ABCC7 p.Pro574His

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PMID: 16442101 [PubMed] Frelet A et al: "Insight in eukaryotic ABC transporter function by mutation analysis."
No. Sentence Comment
307 [138] A455E, P574H cAMP-stimulated apical membrane Cl-currents but current magnitudes were reduced compared to wild-type Electrophysiology of epithelial cells [139] C491S, C1344S, C1355S C491S channels opened almost exclusively to a 3-pS subconductance.
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ABCC7 p.Pro574His 16442101:307:13
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PMID: 10021451 [PubMed] Zeitlin PL et al: "Novel pharmacologic therapies for cystic fibrosis."
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31 As might be expected, mutations in this class, such as R117H or P574H, are thought to confer a milder phenotype. Class V mutations reduce the level of normal CFTR protein by alterations in the promoter or by altering splicing.
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ABCC7 p.Pro574His 10021451:31:64
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131 R347P affects the rate of chloride flow, whereas R117H and P574H reduce the channel open time.
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ABCC7 p.Pro574His 10021451:131:59
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133 Class IV mutations such as R117H, R334W, R347P, A455E, and P574H are associated with a pancreatic sufficient phenotype or late onset pancreatic insufficiency.
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ABCC7 p.Pro574His 10021451:133:59
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PMID: 10362539 [PubMed] Ostedgaard LS et al: "Processing of CFTR bearing the P574H mutation differs from wild-type and deltaF508-CFTR."
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18 Earlier studies showed that the CF-associated mutants, P574H and A455E, were also misprocessed.
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ABCC7 p.Pro574His 10362539:18:55
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19 In this study, we found that processing of P574H and A455E was also temperature-sensitive; at 26°C, some of the protein matured.
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20 In contrast to other CFTR mutants, P574H accumulated in punctate cytoplasmic bodies that colocalized with endoplasmic reticulum (ER) markers.
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ABCC7 p.Pro574His 10362539:20:35
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22 P574H showed a prolonged association with Hsp70 and also colocalized with Hsp70.
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ABCC7 p.Pro574His 10362539:22:0
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24 Unlike wild-type CFTR, which was converted into an intermediate that was stable in the presence of BFA at 37°C, ∆F508 and P574H produced the intermediate only when the temperature was reduced to 26°C. Furthermore the wild-type intermediate was not associated with Hsp70.
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ABCC7 p.Pro574His 10362539:24:134
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26 Key words: Cystic fibrosis, Hsp70, Protein biosynthesis SUMMARY Processing of CFTR bearing the P574H mutation differs from wild-type and ∆F508-CFTR Lynda S. Ostedgaard, Bernhardt Zeiher and Michael J. Welsh* Howard Hughes Medical Institute, Departments of Internal Medicine and Physiology and Biophysics, University of Iowa College of Medicine, Iowa City, Iowa 52242, USA *Author for correspondence Accepted 22 April; published on WWW 10 June 1999 in NBD1 and throughout the protein (Tsui, 1995).
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ABCC7 p.Pro574His 10362539:26:95
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27 We earlier studied two other CF-associated mutations located in NBD1, A455E and P574H (Sheppard et al., 1995).
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29 However, A455E and P574H generate reduced net epithelial current because the proteins are misprocessed and few functional channels reach the plasma membrane (Sheppard et al., 1995; Champigny et al., 1995).
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ABCC7 p.Pro574His 10362539:29:19
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30 Nevertheless, the processing defect of A455E and P574H is less pronounced than that of ∆F508 and the resulting clinical phenotype is less severe (Kristidis et al., 1992; Kerem et al., 1990a; Veeze et al., 1994; Gan et al., 1995).
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ABCC7 p.Pro574His 10362539:30:49
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32 In this study, we compared the processing of P574H and A455E mutants to that of ∆F508.
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ABCC7 p.Pro574His 10362539:32:45
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33 By studying mutants with different degrees of misprocessing, we hope to gain further insight into the biosynthesis of both normal and mutant CFTR which may help design interventions to improve the processing of ∆F508, P574H, and possibly other CF-associated mutants.
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ABCC7 p.Pro574His 10362539:33:225
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37 Using the pcDNA3-6His-CFTR as a backbone, we made the constructs A455E, P574H and ∆F508 (Kunkel, 1985) and confirmed the mutations by DNA sequencing in both directions.
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ABCC7 p.Pro574His 10362539:37:72
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65 RESULTS Because earlier studies suggested that lowering the temperature allowed ∆F508 to fold correctly and exit the ER (Denning et al., 1992a; Lukacs et al., 1993; Sato et al., 1996), we first examined the temperature-sensitivity of A455E and P574H processing.
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ABCC7 p.Pro574His 10362539:65:251
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70 For the NBD1 mutants, ∆F508 (B), A455E (C) and P574H (D), band B was the primary form detected at 37°C.
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ABCC7 p.Pro574His 10362539:70:54
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74 For ∆F508 (B) and A455E (C), the relative amount of band C was minimal at each time at 37°C. Although the relative amount of band C in P574H (D) was also low, it increased slowly with time at 37°C.
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ABCC7 p.Pro574His 10362539:74:147
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75 When the temperature was reduced to 26°C, the relative amount of band C for ∆F508 and A455E increased modestly, while the amount of P574H band C relative to total P574H increased.
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ABCC7 p.Pro574His 10362539:75:144
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76 Although lowering the temperature caused an increase in the relative amount of P574H band C, the total amount of P574H band C was not as high as that in wild-type CFTR.
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ABCC7 p.Pro574His 10362539:76:79
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ABCC7 p.Pro574His 10362539:76:113
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77 These results indicate that, like ∆F508, both A455E and P574H are temperature-sensitive processing mutants.
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ABCC7 p.Pro574His 10362539:77:63
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78 Moreover, P574H makes relatively more band C at both 37°C and 26°C than either ∆F508 or A455E.
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ABCC7 p.Pro574His 10362539:78:10
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82 P574H and A455E are temperature-sensitive mutants.
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ABCC7 p.Pro574His 10362539:82:0
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83 COS-7 cells were electroporated with pcDNA3 vectors encoding wild-type CFTR (A), ∆F508 (B), A455E (C), and P574H (D).
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ABCC7 p.Pro574His 10362539:83:114
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92 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 5 10 15 20 25 30 Chase (hrs) P574H - BFA P574H + BFA ∆F508 - BFA ∆F508 + BFA Wild-type - BFA Wild-type + BFA 0.0 0.2 0.4 0.6 0.8 1.0 1.2 0 5 10 15 20 25 30 Chase (hrs) P574H - BFA P574H + BFA ∆F508 - BFA ∆F508 + BFA A: 37 ˚C B: 26 ˚C RelatoveamountofBandBRelatoveamountofBandB Fig. 2.
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ABCC7 p.Pro574His 10362539:92:59
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ABCC7 p.Pro574His 10362539:92:71
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ABCC7 p.Pro574His 10362539:92:212
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ABCC7 p.Pro574His 10362539:92:224
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93 Effect of brefeldin A on the relative amount of band B protein at 37°C or 26°C. HeLa cells infected with recombinant vaccinia virus encoding P574H-CFTR, ∆F508-CFTR, and wild-type CFTR were pulsed (P574H-CFTR and ∆F508-CFTR for 30 minutes; wild-type-CFTR for 15 minutes) with [35S]methionine at 5 hours post-infection and chased for the indicated times with 10 mM cold methionine in the presence or absence of (5 µg/ml) brefeldin A (BFA).
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ABCC7 p.Pro574His 10362539:93:151
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ABCC7 p.Pro574His 10362539:93:214
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95 Chase was continued for 30 hours for P574H and ∆F508 to detect the stable B form.
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ABCC7 p.Pro574His 10362539:95:37
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99 Repeated measures analysis with multiple means comparison using Supernova software (Abacus Concepts, Berkeley, CA) indicates that P574H + BFA is different from ∆F508 + BFA from 7.5 hours through 30 hours (P=0.013).
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ABCC7 p.Pro574His 10362539:99:130
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100 Wild-type immunoprecipitated with anti-CFTR antibodies (-BFA, ᭺; +BFA, ᭹); ∆F508 (-BFA, ᭝; +BFA, ᭡); P574H (-BFA, ᭛; + BFA, ᭜).
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ABCC7 p.Pro574His 10362539:100:136
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107 Unlike wild-type CFTR, neither ∆F508 nor P574H formed detectable intermediate B after BFA treatment at 37°C (Fig. 2A).
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ABCC7 p.Pro574His 10362539:107:48
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108 However, when the temperature was lowered to 26°C, the intermediate B form of P574H was detectable after 7.5 hours of BFA treatment (Fig. 2B), a time that correlates with the production of P574H band C in the absence of BFA (Fig. 1D).
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ABCC7 p.Pro574His 10362539:108:83
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ABCC7 p.Pro574His 10362539:108:194
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109 Likewise, a small amount of intermediate B accumulated after incubation of ∆F508-expresssing cells at 26°C, but this accumulation of ∆F508 was slower than the accumulation of P574H intermediate B at 26°C (Fig. 2).
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ABCC7 p.Pro574His 10362539:109:194
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110 These data suggest that the temperature-sensitive maturation defect of ∆F508 and P574H occurs at or prior to generation of intermediate band B protein and that P574H responds more readily to lowered temperature than ∆F508.
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ABCC7 p.Pro574His 10362539:110:88
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ABCC7 p.Pro574His 10362539:110:167
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111 We used immunocytochemistry to determine if the cellular distribution of ∆F508, A455E and P574H was consistent with the quantitative biochemical differences we had observed.
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ABCC7 p.Pro574His 10362539:111:97
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117 The NBD1 mutants are temperature-sensitive and P574H displays a unique cytoplasmic pattern of immunofluorescence at 37°C.
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ABCC7 p.Pro574His 10362539:117:47
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118 Immunofluorescence in COS-7 cells electroporated with wild-type-CFTR (A,B); ∆F508 (C,D); A455E (E,F); and P574H (G,H).
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ABCC7 p.Pro574His 10362539:118:113
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120 The same punctate cytoplasmic bodies are present when P574H is expressed in HeLa cells at 37°C (not shown).
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ABCC7 p.Pro574His 10362539:120:54
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121 pattern, P574H presented an unusual immunofluorescence pattern of prominent punctate bodies distributed within the cytoplasm when cells were grown at 37°C (G).
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ABCC7 p.Pro574His 10362539:121:9
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124 We asked whether the cytoplasmic bodies produced by P574H colocalized with a known intracellular organelle.
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ABCC7 p.Pro574His 10362539:124:52
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128 Moreover, in cells stained with both anti-CFTR and anti-Golgi antibodies, the bodies which contain P574H (Fig. 4C) did not colocalize with the Golgi markers, p58 (Bloom and Brashear, 1989) (Fig. 4D) or β-COP (Duden et al., 1991) (not shown).
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ABCC7 p.Pro574His 10362539:128:99
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129 These data suggested the P574H cytoplasmic bodies were not part of the Golgi complex.
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ABCC7 p.Pro574His 10362539:129:25
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130 We used cAMP agonists, which have been shown by others to influence membrane insertion and retrieval of endosomal CFTR (Bradbury et al., 1992; Lehrich et al., 1998), to determine if the P574H bodies were part of endocytic or exocytic vesicles.
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ABCC7 p.Pro574His 10362539:130:186
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132 Although CFTR has also been reported to be contained in clathrin-coated vesicles (Bradbury et al., 1994), the punctate P574H bodies did not colocalize with the more disperse network of clathrin, a component of the trans-Golgi network and the membrane coat of endocytic vesicles and lysosomes (Brodsky, 1988) (not shown).
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ABCC7 p.Pro574His 10362539:132:119
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133 To determine if P574H was present in the ER, we examined the staining pattern of the ER-resident protein, protein disulfide isomerase (PDI) (Kaetzel et al., 1987).
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ABCC7 p.Pro574His 10362539:133:16
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135 Thus P574H, like ∆F508, is retained within the reticular network of the ER.
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ABCC7 p.Pro574His 10362539:135:5
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136 In addition, the punctate P574H cytoplasmic bodies stained with both anti-CFTR and anti-PDI antibodies.
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ABCC7 p.Pro574His 10362539:136:26
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137 These results suggest not only that P574H is localized in the ER, but that the punctate cytoplasmic bodies may represent a subdomain of the ER which is only detectable in cells expressing P574H.
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ABCC7 p.Pro574His 10362539:137:36
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ABCC7 p.Pro574His 10362539:137:188
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138 Because previous work showed that ∆F508 was associated with the cytoplasmic chaperone Hsp70 (Yang et al., 1993), we examined the possibility that the P574H in the punctate bodies might associate with Hsp70.
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ABCC7 p.Pro574His 10362539:138:157
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139 When the same cells are stained with both anti-CFTR and anti-Hsp70 antibodies, P574H (Fig. 5C) and Hsp70 (Fig. 5D) show a striking colocalization in both the reticular ER pattern and in the cytoplasmic bodies.
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ABCC7 p.Pro574His 10362539:139:79
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140 Colocalization of P574H and Hsp70 suggested a physical association of P574H with Hsp70.
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ABCC7 p.Pro574His 10362539:140:18
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141 To test this, we used anti-Hsp70 antibodies to coimmunoprecipitate P574H that was bound to Hsp70 and evaluated the time course of the retention in a pulse-chase experiment.
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ABCC7 p.Pro574His 10362539:141:67
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142 Fig. 6 shows that band B, but not band C, of wild-type CFTR, ∆F508 and P574H were all initially associated with Hsp70.
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ABCC7 p.Pro574His 10362539:142:78
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144 However, band B of both ∆F508 and P574H retain their association with Hsp70 for Fig. 4.
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ABCC7 p.Pro574His 10362539:144:41
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145 P574H cytoplasmic bodies are not part of the Golgi complex.
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146 COS-7 cells expressing P574H and grown at 37°C were either treated with brefeldin A (BFA) (5 µg/ml) (B) or the vehicle control (A) for 30 minutes before staining with anti-CFTR antibody.
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147 Cells expressing P574H grown at 37°C were stained with anti-CFTR antibody (C) and anti-Golgi (p58) antibody (D).
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149 P574H cytoplasmic bodies colocalize with ER markers and with Hsp70.
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150 Cells expressing P574H grown at 37°C were stained with anti-CFTR antibody (A) and with protein-disulphide isomerase antibody (PDI) (B).
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151 Cells expressing P574H grown at 37°C were stained with anti-CFTR antibody (C) and with anti-Hsp70 antibody (D).
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153 In addition, pulse-chase studies show that relatively more of P574H is associated with Hsp70 than is wild-type CFTR (Fig. 6B).
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ABCC7 p.Pro574His 10362539:153:62
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154 This continued physical association of P574H and Hsp70 is consistent with the immunocytochemical colocalization of P574H and Hsp70.
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ABCC7 p.Pro574His 10362539:154:39
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ABCC7 p.Pro574His 10362539:154:115
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158 The continued retention of P574H with Hsp70 suggests that the conformational maturation of P574H may be delayed or actually inhibited, consistent with the diminished ability of P574H to adopt the intermediate B conformation at 37°C.
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ABCC7 p.Pro574His 10362539:158:27
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ABCC7 p.Pro574His 10362539:158:91
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165 DISCUSSION Earlier work showed that CFTR containing the CF-associated mutations ∆F508, A455E, or P574H decreases cell membrane Cl- current primarily because the mutant proteins fail to fold correctly and therefore do not traffic out of the ER (Cheng et al., 1990; Lukacs et al., 1994; Ward and Kopito, 1994; Sheppard et al., 1995; Qu and Thomas, 1996; Qu et al., 1997; Yang et al., 1993; Zhang et al., 1998).
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ABCC7 p.Pro574His 10362539:165:104
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167 The association of P574H and ∆F508 with Hsp70 is prolonged.
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168 HeLa cells infected with recombinant vaccinia virus encoding wild-type-CFTR, ∆F508 and P574H were pulsed for 15 minutes with [35S]methionine at 5 hours post-infection and chased for the indicated times with 10 mM cold methionine at 37°C.
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ABCC7 p.Pro574His 10362539:168:94
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174 Wild-type-CFTR (᭺); P574H (᭛); ∆F508 (᭡).
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185 We found that, like ∆F508, the processing of P574H and A455E is temperature-sensitive.
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ABCC7 p.Pro574His 10362539:185:52
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186 Moreover, unlike ∆F508 and A455E, P574H formed some mature protein at 37°C, and at 26°C, P574H generated relatively more mature protein than ∆F508.
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ABCC7 p.Pro574His 10362539:186:41
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187 Thus the processing defect is less severe for P574H, consistent with functional studies (Sheppard et al., 1995).
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ABCC7 p.Pro574His 10362539:187:46
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188 These studies demonstrate that misprocessing is not an all-or-none phenomenon, but rather a continuum, with wild-type P574H >A455E>>∆F508.
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ABCC7 p.Pro574His 10362539:188:118
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189 This is consistent with the clinical phenotype in CF patients: P574H and A455E are associated with a milder, pancreatic-sufficient phenotype and ∆F508 is associated with a severe, pancreatic-insufficient clinical phenotype (Kerem et al., 1990a,b; Kristidis et al., 1992; Veeze et al., 1994; Gan et al., 1995).
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ABCC7 p.Pro574His 10362539:189:63
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192 This was especially clear for the P574H mutant: the appearance of the intermediate B form in cells treated with BFA occurred at the same time as the maturation of band B to band C in the cells not treated with BFA, suggesting, as has been shown for wild-type, that intermediate B goes on to become the mature band C form of the protein.
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ABCC7 p.Pro574His 10362539:192:34
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193 At 37°C, P574H resides in the ER; P574H showed both the characteristic reticular pattern of immunocytochemical staining typical of ER as well as a unique punctate pattern of cytoplasmic bodies which colocalized with an ER marker.
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ABCC7 p.Pro574His 10362539:193:14
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194 More interestingly, P574H also colocalized with the chaperone Hsp70 in both the reticular ER and in the punctate bodies and was associated with Hsp70 by coimmunoprecipitation.
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ABCC7 p.Pro574His 10362539:194:20
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195 When the temperature was reduced to 26°C, the cytoplasmic bodies containing P574H and Hsp70 were no longer observed, suggested they had dissipated and P574H proceeded to make both the intermediate B and the band C forms of protein.
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ABCC7 p.Pro574His 10362539:195:81
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196 The punctate bodies appeared only in cells expressing P574H.
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198 This morphological pattern is not simply due to the level of protein expression, because the absolute amount of recombinant protein expressed is low and protein expression was no greater for P574H than for any other forms of CFTR.
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ABCC7 p.Pro574His 10362539:198:191
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200 Interestingly, the appearance of subcellular structures reminiscent of the P574H bodies have been reported following expression of other endogenous and recombinant membrane proteins.
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205 We do not know the functional significance of the P574H accumulated within the punctate bodies; it may represent a pool of protein which can convert to the intermediate B form when the temperature is lowered.
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206 The late onset of modest amounts of mature P574H band C at 37°C may represent a slow conversion or 'leak` from this protected pool to intermediate B.
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209 Alternatively, we cannot exclude the possibility that the punctate bodies in cells expressing P574H could represent an ER subcompartment en-route to degradation.
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211 The inability of P574H to form intermediate B and the prolonged retention of P574H by Hsp70 are consistent with this observation.
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ABCC7 p.Pro574His 10362539:211:17
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PMID: 10720936 [PubMed] Zeitlin PL et al: "Pharmacologic restoration of delta F508 CFTR-mediated chloride current."
No. Sentence Comment
106 R347P affects the rate of chloride flow, Chemical and pharmacologic mediators of protein whereas R117H and P574H reduce the channel open trafficking are needed in CF and other diseases caused time.
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ABCC7 p.Pro574His 10720936:106:107
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PMID: 10764788 [PubMed] Van Oene M et al: "Cystic fibrosis mutations lead to carboxyl-terminal fragments that highlight an early biogenesis step of the cystic fibrosis transmembrane conductance regulator."
No. Sentence Comment
1 Analysis of cystic fibrosis-associated missense mutations in the first nucleotide binding domain (NBD1), including A455E, S549R, Y563N, and P574H, revealed reduced levels of mature CFTR with elevated levels of carboxyl-terminal polypeptide fragments of 105 and 90 kDa.
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ABCC7 p.Pro574His 10764788:1:140
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41 EXPERIMENTAL PROCEDURES Construction of CFTR Expression Plasmids-The CFTR mutants A455E, S549R, P574H, and Y563N were generated from pBQ6.2 (34) as described previously (35).
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ABCC7 p.Pro574His 10764788:41:96
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84 Analysis of the S549R mutant showed measurable but intermediate levels of band C, whereas A455E, Y563N, and P574H mutants showed markedly reduced levels using both tagged and untagged CFTR.
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ABCC7 p.Pro574His 10764788:84:109
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238 The A455E, Y563N, and P574H mutations do appear to be able to achieve at least nominal levels of chloride conduction at the cell surface based both on the presenting phenotype in CF patients (54) and on single channel and whole cell current measurements (55) in heterologous expression systems.
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ABCC7 p.Pro574His 10764788:238:22
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239 In contrast to the Y563N and P574H mutations, where low levels of mature band C forms were detectable with long exposure and/or long label incorporation times in HEK293 cells (data not shown), fully glycosylated protein could not be detected for A455E using our assay systems.
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ABCC7 p.Pro574His 10764788:239:29
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PMID: 10940786 [PubMed] Zeitlin PL et al: "Future pharmacological treatment of cystic fibrosis."
No. Sentence Comment
22 Examples of CFTR mutations organized by classification of the defect in CFTR biosynthesis Type Genotype Phenotype Defect Cell diagram Drugs that may improve phenotype G542X 621+1 G → T 3905insT W1282X R553X 1717-1 G → A PI no CFTR protein no cell surface chloride transport gentamicin G418 Class II [64] 'F508 N1303K (P574H)a (A455E)a PI defective CFTR processing defective CFTR trafficking no cell surface chloride transport chemical chaperones CPX phenylbutyrate deoxyspergualin Class III [64] G551D G551S PI defective chloride channel regulation reduced or absent cell surface chloride transport genistein pyrophosphate Class IV [64, 66] R117H R334W G314E R347P ('F508)a P574H PS reduced chloride conductance reduced levels of cell surface chloride transport genistein milrinone phenylbutyrate Class V [64] 3849+10 kb C → T 2789+5 G → A 3272-26 A → G A455E 3120+1 G → A 1811+1.6 kb A → G 5Tb PS normal CFTR channels reduced numbers of normal CFTR reduced cell surface chloride transport genistein milrinone phenylbutyrate a Some mutants have features of more than one class of defect.
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ABCC7 p.Pro574His 10940786:22:332
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ABCC7 p.Pro574His 10940786:22:688
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75 Another misprocessed mutant, P574H, has been shown to associate with Hsp70 in a prolonged state unless the temperature of the culture was reduced to 26°C [48].
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ABCC7 p.Pro574His 10940786:75:29
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PMID: 11100963 [PubMed] Choo-Kang LR et al: "Type I, II, III, IV, and V cystic fibrosis transmembrane conductance regulator defects and opportunities for therapy."
No. Sentence Comment
108 R347P affects the rate of chloride flow, whereas R117H and P574H reduce the channel open time.
X
ABCC7 p.Pro574His 11100963:108:59
status: NEW
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124 Missense mutations that belong to this class include P574H and A455E.
X
ABCC7 p.Pro574His 11100963:124:53
status: NEW
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PMID: 11116277 [PubMed] Roomans GM et al: "Pharmacological treatment of the ion transport defect in cystic fibrosis."
No. Sentence Comment
199 A substituted benzimidazolone, NS-004, was shown to activate wild-type CFTR and ∆F508-CFTR chloride channels in Xenopus oocytes and Vero cells expressing these channels [105] and also restored near-normal CFTR activity in cells expressing the P574H-CFTR channel (a mild mutation of CF) [106].
X
ABCC7 p.Pro574His 11116277:199:250
status: NEW
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PMID: 11388756 [PubMed] Heim RA et al: "Improved detection of cystic fibrosis mutations in the heterogeneous U.S. population using an expanded, pan-ethnic mutation panel."
No. Sentence Comment
85 These mutations, 574delA, C524X, Y563D, P574H, 2043delG, 3662delA, 3821delT, Q1238X, and 3849 ϩ 4AϾG, as well as the previous seven mutations, make no detectable contribution to mutation detection in the general U.S. CF population.
X
ABCC7 p.Pro574His 11388756:85:40
status: NEW
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PMID: 11404246 [PubMed] Choo-Kang LR et al: "Induction of HSP70 promotes DeltaF508 CFTR trafficking."
No. Sentence Comment
281 Whether this approach would also overcome the defects caused by other trafficking mutants such as N1303K, P574H, or G480C remains to be tested.
X
ABCC7 p.Pro574His 11404246:281:106
status: NEW
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PMID: 11589722 [PubMed] Walkowiak J et al: "Analysis of exocrine pancreatic function in cystic fibrosis: one mild CFTR mutation does not exclude pancreatic insufficiency."
No. Sentence Comment
86 Kristidis et al. [10] reported that pancreatic insufficiency strongly correlates also with two alleles of DI507, Q493X, G542X, R553X, W1282X, 621 1 1G-T, 1717±1G-A, 556delA, 3659delC, I148T, G480C, V520F and R560T while one or two mutations such as R117H, R334W, A455E, and P574H were correlated with a pancreatic sufficient phenotype.
X
ABCC7 p.Pro574His 11589722:86:279
status: NEW
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PMID: 11966405 [PubMed] Sangiuolo F et al: "Towards the pharmacogenomics of cystic fibrosis."
No. Sentence Comment
111 Gentamicin Neomicin (G418) Class II Mutations that fail to be properly processed to a matureglycosylatedform and transported to the apical membrane ∆F508 N1303K P574H A455E PI Defective CFTR processing and trafficking.
X
ABCC7 p.Pro574His 11966405:111:168
status: NEW
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114 R117H R334W G314E R347P ∆F508 P574H PS Reduced chloride conductance Reduced levels of cell surface chloride transport Genistein Milrinone Phenylbutyrate UTP INS36217 Moli1901 Class V Mutations causing defects in CFTR channel expression levels.
X
ABCC7 p.Pro574His 11966405:114:37
status: NEW
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PMID: 12124743 [PubMed] Salvatore F et al: "Genotype-phenotype correlation in cystic fibrosis: the role of modifier genes."
No. Sentence Comment
46 A series of mutations usually associated with pancreatic sufficiency have been identified and defined as ''mild`` with reference to pancreatic status [Kerem et al., 1989c]: G85E, G91R, R117H, E193K, P205S, R334W, T338I, R347H, R347L, R347P, R352Q, A455E, S492F, S549N, P574H, D579G, 711 þ 5 G > A, C866Y, F1052V, H1054D, R1066H, R1068H, H1085R, D1152H, S1159P, S1251N, F1286S, G1349D, 2789 þ 5 G > A, and 3849 þ 10kb C > T [Dean et al., 1990; Cutting et al., 1990a; Cremonesi et al., 1992; Highsmith et al., 1994].
X
ABCC7 p.Pro574His 12124743:46:269
status: NEW
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PMID: 12815607 [PubMed] Scotet V et al: "Comparison of the CFTR mutation spectrum in three cohorts of patients of Celtic origin from Brittany (France) and Ireland."
No. Sentence Comment
64 Spectrum of the CFTR Mutations Identified in the Cohorts from Brittany, Dublin Centre, and Cork Area Nucleotide Amino acid change * change Exon Number Frequency Number Frequency Number Frequency 211delG 2 1 0.1% 310G>T E60X 3 5 0.6% 4 0.3% 347C>A A72D 3 1 0.1% 368G>A W79X 3 1 0.1% 386G>A G85E 3 2 0.3% 3 0.2% 403G>A G91R 3 2 0.3% 482G>A R117H 4 4 0.5% 38 3.0% 4 1.4% 498T>A Y122X 4 1 0.1% 574delA 4 1 0.1% 577G>A G149R 4 1 0.1% 621+1G>T int 4 5 0.6% 21 1.7% 790C>T Q220X 6a 1 0.1% 875+1G>C int 6a 1 0.4% 905delG 6b 1 0.1% 1065C>G F311L 7 2 0.3% 1078delT 7 28 3.6% 1132C>T R334W 7 1 0.1% 1172G>A R347H 7 5 0.6% 1172G>T R347L 7 1 0.1% 1172G>C R347P 7 1 0.1% 1187G>A R352Q 7 3 0.2% 2 0.7% 1208A>G Q359R 7 1 0.1% 1154insTC 7 2 0.2% 1221delCT 7 2 0.3% 1248+1G>A int 7 1 0.1% 1249-27delTA int 7 1 0.4% 1334G>A W401X 8 1 0.1% 1461ins4 9 5 0.4% 1471delA 9 2 0.2% 1607C>T S492F 10 2 0.3% 1609C>T Q493X 10 1 0.1% 1648_1653delATC I507del 10 3 0.4% 10 0.8% 1 0.4% 1652_1655del 3 bp F508del 10 582 74.8% 966 76.5% 226 81.3% 1690G>T V520F 10 4 0.3% 1717-1G>A int 10 8 1.0% 9 0.7% 1756G>T G542X 11 5 0.6% 8 0.6% 1779T>G S549R 11 1 0.1% 1784G>A G551D 11 29 3.7% 82 6.5% 27 9.7% 1789C>G R553G 11 1 0.1% 1789C>T R553X 11 3 0.4% 1 0.1% 1806delA 11 1 0.1% 1811G>A R560K 11 2 0.3% 1811G>C R560T 11 30 2.4% 2 0.7% 1819T>A Y563N 12 1 0.1% 1853C>A P574H 12 1 0.1% 1898+1G>A int 12 1 0.1% 2184delA 13 1 0.1% 1 0.1% 2184insA 13 1 0.1% 2622+1G>A int 13 1 0.1% 2 0.2% 2622+1G>T int 13 1 0.1% 2623-2A>G ** int 13 1 0.1% 2670G>A W846X2 14a 8 1.0% 2752-1G>T int 14a 1 0.1% 2752-26A>G int 14a 2 0.2% 2789+5G>A int 14b 6 0.8% 2966C>T S945L 15 2 0.3% 3007delG 15 4 0.3% 3040G>C G970R 15 1 0.1% 3062C>T S977F 16 1 0.1% 3120+1G>A int 16 1 0.1% 3272-26A>G int 17a 4 0.5% 2 0.2% 2 0.7% 3320dupli(CTATG) 17b 1 0.1% 3329G>A R1066H 17b 1 0.1% 3340C>T R1070W 17b 1 0.1% 3408C>A Y1092X 17b 7 0.9% 3442G>T E1104X 17b 1 0.1% 3446T>G ** M1105R 17b 1 0.1% 3586G>C D1152H 18 1 0.1% 3601-17T>C + 1367delC int 18 + 9 1 0.1% 3616C>T R1162X 19 1 0.1% 2 0.2% 3659delC 19 2 0.2% 3832A>G I1234V 19 2 0.3% 3849+4A>G int 19 1 0.1% 3849+10kbC>T int 19 3 0.2% 3877G>A G1249R 20 1 0.1% 3884G>A S1251N 20 1 0.1% 3898insC 20 1 0.1% 3905insT 20 2 0.3% 3978G>A W1282X 20 3 0.4% 4005+1G>A int 20 6 0.8% 4016insT 21 1 0.1% 4041C>G N1303K 21 11 1.4% 5 0.4% 4136T>C L1335P 22 1 0.1% 1 0.4% 4279insA 23 1 0.1% Unidentified Unidentified - 3 0.4% 41 3.2% 11 4.0% Total 778 100.0% 1262 100.0% 278 100.0% * All nucleotide changes correspond to cDNA numbering.
X
ABCC7 p.Pro574His 12815607:64:1325
status: NEW
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PMID: 12865275 [PubMed] Ahmed N et al: "Molecular consequences of cystic fibrosis transmembrane regulator (CFTR) gene mutations in the exocrine pancreas."
No. Sentence Comment
309 Table 2 Genotype classification according to the functional consequences of CFTR gene mutations Pancreatic status Class I Class II Class III Class IV Class V PS F1 , 875+1G→C(2) F, F (1) F, G551D (1) F, R117H (11) F,3849+10kbC→T (5) F, G85E2 (1) F, R347H (3) F,3272-26A→G (4) F, S1251N (2) F,A445E (3) F, D614G (1) F,P574H (2) F, R347P (1) F,3120G>A (1) R117H,R117H (1) F, 5T (8) F, L1335P (1) F,2789+5G→A (1) F,P67L (1) F,R347P/R347H (1) F,V232D(2) R334W, R334W(1) PS→PI F,3659delC (1) F,F (15) F,G551D (1) F, I1234V (1) F,2184insA (1) F,R560T (1) PI F, G542X (27) F,F (365) F, G551D (28) F, 621+1G→T (13) F, R560T (7) F,R553X (7) F, N1303K (9) F, R1162X (6) F,L1077P (2) F, 3659delC (5) F, I48T (1) F, 1717-1G→A (5) F,A559T (1) F, W1282X (5) F, G85E2 (2) F, 711+1G→T (5) G551D,G551D(1) F,2184delA(4) F,H199R (1) W1282X,W1282X (4) F,I1072T(1) F,Y1092X (3) F,S549 (R75Q) (1) F,556delA (3) F, Q493X (3) F,4016InsT (3) F, 3120+1G→A (2) F, G551D/R553X (2) F,Q814X(2) F,1154insTC (2) F,441delA (1) F, 4326delTC (1) F,Q552X(1) F,3007delG (1) F,2184insA (1) F, 4010del4 (1) F,3905insT (1) F,1078delT(1) F,E1104X (1) F,3876delA (1) F,4374+1G→T (1) F,E585X (1) F, E60X (1) CFTR, cystic fibrosis transmembrane regulator; PI, pancreatic insufficiency; PS, pancreatic sufficiency.
X
ABCC7 p.Pro574His 12865275:309:338
status: NEW
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PMID: 14502435 [PubMed] Derand R et al: "Comparative pharmacology of the activity of wild-type and G551D mutated CFTR chloride channel: effect of the benzimidazolone derivative NS004."
No. Sentence Comment
204 Further studies have also demonstrated that NS004 is a modulator of P574H (Champigny et al., 1995), K1250A (Al-Nakkash et al., 2001) and delF508 (Gribkoff et al., 1994; He et al., 1998; Al-Nakkash et al., 2001).
X
ABCC7 p.Pro574His 14502435:204:68
status: NEW
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PMID: 14562574 [PubMed] Morinville V et al: "Genetic disorders of the pancreas."
No. Sentence Comment
84 Mutations of CFTR include: deltaF508, R117H, D1152H, P574H, 3120 G > A, 621 + 1 G > T, G1069R, N1303K.
X
ABCC7 p.Pro574His 14562574:84:53
status: NEW
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PMID: 14586256 [PubMed] Reboul MP et al: "Isolated idiopathic chronic pancreatitis associated with a compound heterozygosity for two mutations of the CFTR gene."
No. Sentence Comment
80 Patient CFTR no PolyT genotype Sex genotype Age (years) Sweat chloride (mmol/L) Anamnestic features known to be associated with atypical CF Reference 1 F508del/R117H 9T/7T M 45 29 CBAVD [4] 2 N1303K/R117H 9T/7T F n.a. 37 bronchiectasis, sinusitis, positive NPD [5] 3 R1162X/2789+5G>A 7T/7T F n.a. 108 chronic cough [5] 4 I336K/R75Q 7T/7T F 26 26 nasal polyposis [7] 5 F508del/L997F 9T/7T M 17 24 none [11] 6 3849+10kbC>T/3878delG 7T/7T M 14 n.a. none [11] 7 S1235R/L997F 5T/7T M 27 25 none [11] 8 F508del/R117H n.a. M 45 29 CBAVD, smooth P. aeruginosa [12] 9 F508del/I1027T n.a. F 32 59 none [12] 10 F508del/D1152H n.a. M 8 62 none [12] 11 F508del/D1152H n.a. F 15 32 none [12] 12 F508del/P574H n.a. F 26 81 sinus surgery, S. aureus, S. maltophilia [12] 13 F508del/3120G>A n.a. F 40 n.a. n.a. [12] 14 F508del/G1069R n.a. M 16 n.a. n.a. [12] 15 G542X/S1235R 7T/7T M 35 15 none [this study] n.a.: not available.
X
ABCC7 p.Pro574His 14586256:80:689
status: NEW
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PMID: 14641997 [PubMed] Raskin S et al: "High allelic heterogeneity between Afro-Brazilians and Euro-Brazilians impacts cystic fibrosis genetic testing."
No. Sentence Comment
63 FREQUENCIES OF 70 CFTR MUTATIONS IN DIFFERENT STATES OF BRAZIL, BY CONTINENTA L GROUP CFTR mutations SC PR MG detected n n n n % n % N % DF508 53 39 54 146 47.1 8 10.5 154 39.9 G542X 6 9 8 23 7.4 1 1.3 24 6.2 R1162X 9 2 4 15 4.8 2 2.6 17 4.4 N1303K 5 5 0 10 3.2 0 0 10 2.6 R334W 5 1 4 10 3.2 0 0 10 2.6 G85E 2 2 4 8 2.6 1 1.3 9 2.3 1717-1G®A 1 3 2 6 1.9 0 0 6 1.6 W1282X 4 1 1 6 1.9 0 0 6 1.6 3849110kbC®T 1 3 1 5 1.6 0 0 5 1.3 R553X 0 2 0 2 0.7 0 0 2 0.5 1812-1G®A 0 1 3 4 1.3 1 1.3 5 1.3 2183AA®G 2 1 0 3 1.0 0 0 3 0.8 312011G®A 0 0 2 2 0.7 2 2.6 4 1.0 Y1092X 0 1 1 2 0.7 1 1.3 3 0.8 G551D 0 0 0 0 0 0 0 0 0 W1089X 0 0 1 1 0.3 0 0 1 0.3 6211G®T 0 1 0 1 0.3 0 0 1 0.3 Q1238X 0 1 0 1 0.3 0 0 1 0.3 711-1G®T 0 1 0 1 0.3 0 0 1 0.3 R347P 1 0 0 1 0.3 0 0 1 0.3 189811G®A 1 0 0 1 0.3 0 0 1 0.3 I507 0 0 1 1 0.3 0 0 1 0.3 Subtotal 91 73 86 250 80.7 16 21.1 266 68.9 Alleles with CFTR 5 27 28 60 19.4 60 79.0 120 31.1 mutations not detected Total 96 100 114 310 100.0 76 100.0 386 100.0 Detection rate (%) 94.8 73.0 75.4 250 80.7 16 21.1 266 68.9 The following 70 CFTR mutations were selected and tested on the basis of frequency in various populations, known association with CF, or predicted deleterious effect on the CFTR protein product; DF508, G542X, N1303K, G551D, R553X, DI507, A455E, A559T, C524X, D1270N, E60X, G178R, G330X, G85E, 2307insA, I148T, K710X, P574H, Q1238X, Q493X, Q890X, R1158X, R1162X, R117H, R334W, R347H, R347P 2307insA, I148T, K710X, P574H, Q1238X, Q493X, Q890X, R1158X, R1162X, R117H, R334W, R347H, R347P 2307insA, 1148T, K710X, P574H, Q1238X, Q493X, Q890X, R1158X, R1162X, R117H, R334W, R347H, R347P, R352Q, R560T, S1196X, S1255X, S364P, S549N, S549R, V520F, W1089X, W1282X, W1310X, W1316X, Y1092X, Y122X, Y563D, 1078delT,1677delTA,1717-1G-A,1812-1G-A,1898 1 1G-A, 2043delG,2183delAA-G, 2184delA, 2789 1 5G-A, 2869insG, 2909delT, 3120 1 1G-A, 3120G-A, 3358delAC, 3659delC, 3662delA, 3750delAG, 3791delC, 3821delT, 3849 1 10KbC-T, 3849 1 4A-G, 3905insT, 405 1 1G-A, 444delA, 556delA, 574delA, 621 1 1G-T, and 711 1 1G-T. aSC, Santa Catarina State; PR, Parana State; MG, Minas Gerais State; n, number of chromosomes.
X
ABCC7 p.Pro574His 14641997:63:1397
status: NEW
X
ABCC7 p.Pro574His 14641997:63:1493
status: NEW
X
ABCC7 p.Pro574His 14641997:63:1589
status: NEW
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PMID: 14662004 [PubMed] Zeitlin PL et al: "Emerging drug treatments for cystic fibrosis."
No. Sentence Comment
53 The ∆F508 mutation and others such as N1303K or P574H [26], which are also misprocessed in the ER, have been grouped together as Class 2 mutations in CFTR [27].
X
ABCC7 p.Pro574His 14662004:53:55
status: NEW
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88 Class of mutation Molecular mechanism Pancreatic status (if known) Examples 1 No CFTR protein synthesis PI W1282X, G542X, R553X, 621 + 1 G→T, 1717-1 G→A, 3905insT, 394delTT 2 Abnormal CFTR processing and trafficking PI ∆F508, N1303K, P574H 3 Defective CFTR regulation (normal trafficking) PI G551D, G551S, G1349D, S1255P 4 Decreased CFTR chloride conductance PS R117H, R334W, R347P, P547H 5 Reduced synthesis and trafficking of normal CFTR PS A455E, 3849 + 10kb C→T, (5T) 6A Reduced apical stability PI S1455X, Q1412S, 4326delTC, 4279insA 6B Defective regulation of other ion channels PI G551D Note that the G551D is placed in Class 3 for defective regulation and Class 6B for defective regulation of the outwardly rectifying chloride channel.
X
ABCC7 p.Pro574His 14662004:88:255
status: NEW
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301 OSTEDGAARD LS, ZEIHER B, WELSH MJ: Processing of CFTR bearing the P574H mutation differs from wild-type and deltaF508-CFTR.
X
ABCC7 p.Pro574His 14662004:301:66
status: NEW
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PMID: 15233679 [PubMed] Choudari CP et al: "Risk of pancreatitis with mutation of the cystic fibrosis gene."
No. Sentence Comment
130 In a study of more than 500 CF patients, five mutations (R117H, R334W, R347P, A455E, and P574H) were found exclusively in pancreatic sufficient patients (8).
X
ABCC7 p.Pro574His 15233679:130:89
status: NEW
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PMID: 15274098 [PubMed] Davis PB et al: "Relation of sweat chloride concentration to severity of lung disease in cystic fibrosis."
No. Sentence Comment
27 T; G91R; E92K; P205S; G551S; Y563N; and P574H.23,24 Note that there are 36 mild alleles in 34 subjects, because two subjects had both the 3848 þ 10 kb C !
X
ABCC7 p.Pro574His 15274098:27:40
status: NEW
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PMID: 15343184 [PubMed] Borowitz D et al: "Use of fecal elastase-1 to classify pancreatic status in patients with cystic fibrosis."
No. Sentence Comment
116 FE-1 values in subjects with CFTR mutations associated with pancreatic sufficiency11 N Mean (mg/g stool) Median (mg/g stool) Range (mg/g stool) Subjects with at least one PS allele* FE-1 >200 mg/g stool 16 584 582.9 349-773 FE-1 <200 mg/g stool 5 64.4 74.8 0-125 Subjects with at least one PS variable alleley FE-1>200 mg/g stool 29 496.2 493.6 224-798 FE-1 <200 mg/g stool 13 76.1 65.9 0-187 *Pancreatic sufficient dominant CF alleles G551S R117H R347H P574H R334W R352Q T3381 yVariable pancreatic sufficient CF mutations G85E 3849 + 10 kb C fi T R347P 2789 + 5G fi A A455E In summary, FE-1 is an accurate, easily obtained screening test to classify patients with CF as PI or PS.
X
ABCC7 p.Pro574His 15343184:116:454
status: NEW
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PMID: 15371903 [PubMed] Sugarman EA et al: "CFTR mutation distribution among U.S. Hispanic and African American individuals: evaluation in cystic fibrosis patient and carrier screening populations."
No. Sentence Comment
35 87 mutation panel The following mutations were included in the panel: ⌬F508, ⌬F311, ⌬I507, A455E, A559T, C524X, D1152H, D1270N, E60X, G178R, G330X, G480C, G542X, G551D, G85E, G91R, I148T, K710X, L206W, M1101K, N1303K, P574H, Q1238X, Q359K/T360K, Q493X, Q552X, Q890X, R1066C, R1158X, R1162X, R117C, R117H, R1283M, R334W, R347H, R347P, R352Q, R553X, R560T, S1196X, S1251N, S1255X, S364P, S549I, S549N, S549R, T338I, V520F, W1089X, W1282X, Y1092X, Y563D, 1078delT, 1161delC, 1609delCA, 1677delTA, 1717-1GϾA, 1812-1GϾA, 1898ϩ1GϾA, 1898ϩ5GϾT, 1949del84, 2043delG, 2143delT, 2183delAAϾG, 2184delA, 2307insA, 2789ϩ5GϾA, 2869insG, 3120ϩ1GϾA, 3120GϾA, 3659delC, 3662delA, 3791delC, 3821delT, 3849ϩ10kbCϾT, 3849ϩ4AϾG, 3905insT, 394delTT, 405ϩ1GϾA, 405ϩ3AϾC, 444delA, 574delA, 621ϩ1GϾT, 711ϩ1GϾT, 711ϩ5GϾA, 712-1GϾT, 3876delA CFTR mutation analysis Genomic DNA was extracted from peripheral blood lymphocytes, buccal cell swabs, or bloodspots by Qiagen QIAmp 96 DNA Blood Kit. Specimens were tested for 87 mutations by a pooled allele-specific oligonucleotide (ASO) hybridization method as previously described.16,17 Two multiplex chain reactions (PCR) were used to amplify 19 regions of the CFTR gene.
X
ABCC7 p.Pro574His 15371903:35:239
status: NEW
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PMID: 15479737 [PubMed] Wang X et al: "COPII-dependent export of cystic fibrosis transmembrane conductance regulator from the ER uses a di-acidic exit code."
No. Sentence Comment
403 Processing of CFTR bearing the P574H mutation differs from wild-type and ⌬F508-CFTR.
X
ABCC7 p.Pro574His 15479737:403:31
status: NEW
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PMID: 15528020 [PubMed] Cohn JA et al: "The role of cystic fibrosis gene mutations in determining susceptibility to chronic pancreatitis."
No. Sentence Comment
74 When these six nominal CF carriers were tested further by DNA sequencing, rare mutations were identified in five cases (D1152H, P574H, G1069R, 3120G > A; each detected rare mutation is mild-variable [18,30,48,49].
X
ABCC7 p.Pro574His 15528020:74:128
status: NEW
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78 The European data Table 1 Abnormal CFTR and PSTI genotypes detected in two studies of idiopathic chronic pancreatitis* CFTR genotype category N Genotypes detected in individual subjects US study (Noone et al [47]) CFsev / CFm-v compound heterozygotes 8 DF508 / R117H-7T**; DF508 / 5T; DF508 / 5T; DF508 / D1152H; DF508 / D1152H; DF508 / P574H; DF508 / 3120G>A; 621þ1G>T/G1069R CFm-v / CFm-v compound heterozygotes 1 5T / 5T** CFsev / - (CF carriers) 1 N1303K / - CFm-v / - 7 R117H-7T / -; 5T / -**; 5T / -; 5T / -; 5T / -; 5T / -; 5T / - Normal (- / -) CFTR genotype 22 1 was homozygous for the N34S PSTI mutation; 5 were N34S carriers European study (Audrezet et al [50]) CFsev / CFm-v compound heterozygotes 4 DF508/R352Q; DF508/P5L; DF508/Q1476X; W1282X/5T*** CFm-v / CFm-v compound heterozygotes 2 V562I/5T; E217G/A1136T CFsev / - (CF carriers)**** 3 DF508 / -; DF508 / -; G542X / - CFm-v / - 9 L967S/-**; IVS18-20T>C/-**; c.4575þ2G>A/-; IVS3-6T>C; 5T/-; 5T/-; 5T/-; 5T/-; 5T/- Normal (- / -) CFTR genotype 17 1 was homozygous for the N34S PSTI mutation; 1 was a N34S carrier * CFTR mutations were classified as causing either severe (CFsev ) or mild-variable loss-of-function (CFm-v ) [18,47]; all detected CFsev mutations are CF-causing mutations according to current consensus criteria [79].
X
ABCC7 p.Pro574His 15528020:78:337
status: NEW
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PMID: 15705292 [PubMed] Claustres M et al: "Molecular pathology of the CFTR locus in male infertility."
No. Sentence Comment
190 However a single mutation can cause more than one type of abnormality, and hence fall into multiple classes (for example, missense P574H is both a class 2 and class 4 mutation, missense G.'S.
X
ABCC7 p.Pro574His 15705292:190:131
status: NEW
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PMID: 15749233 [PubMed] Cohn JA et al: "The impact of cystic fibrosis and PSTI/SPINK1 gene mutations on susceptibility to chronic pancreatitis."
No. Sentence Comment
77 When these six nominal CF carriers were tested further by DNA sequencing, rare mutations were identified in five cases (D1152H, P574H, G1069R, 3120G > A; each detected rare mutation is mild-variable [18,30,48,49].
X
ABCC7 p.Pro574His 15749233:77:128
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90 Table 1 Abnormal CFTR and PSTI genotypes detected in two studies of idiopathic chronic pancreatitis* CFTR genotype category N Genotypes detected in individual subjects US study (Noone et al [47]) CFsev / CFm-v compound heterozygotes 8 DF508 / R117H-7T**; DF508 / 5T; DF508 / 5T; DF508 / D1152H; DF508 / D1152H; DF508 / P574H; DF508 / 3120G>A; 621þ1G>T/G1069R CFm-v / CFm-v compound heterozygotes 1 5T / 5T** CFsev / - (CF carriers) 1 N1303K / - CFm-v / - 7 R117H-7T / -; 5T / -**; 5T / -; 5T / -; 5T / -; 5T / -; 5T / - Normal (- / -) CFTR genotype 22 1 was homozygous for the N34S PSTI mutation; 5 were N34S carriers European study (Audrezet et al [50]) CFsev / CFm-v compound heterozygotes 4 DF508/R352Q; DF508/P5L; DF508/Q1476X; W1282X/5T*** CFm-v / CFm-v compound heterozygotes 2 V562I/5T; E217G/A1136T CFsev / - (CF carriers)**** 3 DF508 / -; DF508 / -; G542X / - CFm-v / - 9 L967S/-**; IVS18-20T>C/-**; c.4575þ2G>A/-; IVS3-6T>C; 5T/-; 5T/-; 5T/-; 5T/-; 5T/- Normal (- / -) CFTR genotype 17 1 was homozygous for the N34S PSTI mutation; 1 was a N34S carrier * CFTR mutations were classified as causing either severe (CFsev ) or mild-variable loss-of-function (CFm-v ) [18,47]; all detected CFsev mutations are CF-causing mutations according to current consensus criteria [79].
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ABCC7 p.Pro574His 15749233:90:319
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PMID: 15758625 [PubMed] Turcios NL et al: "Cystic fibrosis: an overview."
No. Sentence Comment
55 Pancreatic Sufficient CF Mutations Dominant Pancreatic-Sufficient Variable Pancreatic-Sufficient G551S G85E P574H R347P R117H 3849 + 10kb C !
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ABCC7 p.Pro574His 15758625:55:108
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PMID: 15758663 [PubMed] Cohn JA et al: "Reduced CFTR function and the pathobiology of idiopathic pancreatitis."
No. Sentence Comment
51 nominal CF carriers were further tested by DNA sequencing, rare mutations were identified in 5 cases (D1152H, P574H, G1069R, 3120G.A); each detected rare mutation is mild to variable.18,30,48,49 Among the 9 compound heterozygotes, only one would have been corrected classified by CF carrier screening (1).
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ABCC7 p.Pro574His 15758663:51:110
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69 Abnormal CFTR and PSTI Genotypes Detected in Two Studies of ICP CFTR Genotype Category* N Genotypes Detected in Individual Subjects U.S. study (Noone et al47 ) CFsev / CFm-v compound heterozygotes 8 DF508 / R117H-7T †; DF508 / 5T; DF508 / 5T; DF508 / D1152H; DF508 / D1152H; DF508 / P574H; DF508 / 3120G.A; 621 + 1G.T/G1069R CFm-v / CFm-v compound heterozygotes 1 5T / 5T † CFsev / 2 (CF carriers) 1 N1303K / 2 CFm-v / 2 7 R117H-7T / 2; 5T / 2 †; 5T / 2; 5T / 2; 5T / 2; 5T / 2; 5T / 2 Normal (2 / 2) CFTR genotype 22 1 was homozygous for the N34S PSTI mutation; 5 were N34S carriers French study (Audrezet et al50 ) CFsev / CFm-v compound heterozygotes 4 DF508/R352Q; DF508/P5L; DF508/Q1476X; W1282X/5T‡ CFm-v / CFm-v compound heterozygotes 2 V562I/5T; E217G/A1136T CFsev / 2 (CF carriers)§ 3 DF508 / 2; DF508 / 2; G542X / 2 CFm-v / 2 9 L967S/2 †; IVS18-20T.C/ 2†; c.4575+2G.A/2; IVS3-6T.C; 5T/2; 5 /2; 5T/ 2; 5T/2; 5T/ 2 Normal (2 / 2) CFTR genotype 17 1 was homozygous for the N34S PSTI mutation; 1 was a N34S carriers *Mutations of the cystic fibrosis (CF) gene (CFTR) were classified as causing either severe (CFsev ) or mild-variable loss-of-function (CFm-v )18,47 ; all detected CFsev mutations are CF-causing mutations according to current consensus criteria.68 In the U.S. study, most patients were tested for rare mutations by DNA sequencing47 ; in the French study, most patients were tested by dHPL.50 †These patients were also carriers for the N34S mutation of a trypsin inhibitor gene (PSTI).
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ABCC7 p.Pro574His 15758663:69:290
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PMID: 15880796 [PubMed] Kerem E et al: "Pharmacological induction of CFTR function in patients with cystic fibrosis: mutation-specific therapy."
No. Sentence Comment
211 IBMX produced a small, CFTR-related secretory response in the jejunum, cecum, and rectum of G551D mice but had no effect in the nasal epithelium.78 NS-004 restored near-normal channel activity from P574H-CFTR (a mild, trafficking-impaired mutationinNBD1).79 Theseresultssuggestthat,inaddition to their direct effects on CFTR, these drugs may also have other effects that increase the number of channel proteins found in the membrane.
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ABCC7 p.Pro574His 15880796:211:198
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PMID: 16127463 [PubMed] Pedemonte N et al: "Small-molecule correctors of defective DeltaF508-CFTR cellular processing identified by high-throughput screening."
No. Sentence Comment
9 The bisaminomethylbithiazoles corrected ∆F508-CFTR in ∆F508/∆F508 human bronchial epithelia but did not correct a different temperature-sensitive CFTR mutant (P574H-CFTR) or a dopamine receptor mutant.
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ABCC7 p.Pro574His 16127463:9:180
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115 As an initial test of compound specificity for correction of defective ∆F508-CFTR processing, compounds were tested on P574H-CFTR, a mutant that like ∆F508-CFTR is retained at the ER but can be rescued by incubation for 24 hours at reduced temperature (26).
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ABCC7 p.Pro574His 16127463:115:126
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119 Figure 5A shows that corr-4a and -2a, which produced robust chloride currents in ∆F508-CFTR-expressing cells, did not produce significant correction in P574H-CFTR cells, despite a positive low-temperature rescue control.
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ABCC7 p.Pro574His 16127463:119:159
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179 (A) Apical membrane chloride current in FRT cells expressing the temperature-sensitive mutant P574H-CFTR.
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ABCC7 p.Pro574His 16127463:179:94
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193 The lack of effect of some ∆F508-CFTR correctors on the temperature-sensitive mutant P574H-CFTR and the mutant DRD4-M345T suggests their specificity for the ∆F508 mutation, though further studies are needed to evaluate the numerous potential off-target specificities.
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ABCC7 p.Pro574His 16127463:193:92
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202 A similar approach was used to generate BHK cells coexpressing ∆F508-CFTR and YFP-H148Q/I152L and FRT cells stably expressing P574H-CFTR.
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ABCC7 p.Pro574His 16127463:202:133
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365 Processing of CFTR bearing the P574H mutation differs from wild-type and ∆F508-CFTR.
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ABCC7 p.Pro574His 16127463:365:31
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358 Processing of CFTR bearing the P574H mutation differs from wild-type and ∆F508-CFTR.
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ABCC7 p.Pro574His 16127463:358:31
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PMID: 16132229 [PubMed] Eudes R et al: "Nucleotide binding domains of human CFTR: a structural classification of critical residues and disease-causing mutations."
No. Sentence Comment
131 Other examples (fig. 1) are: (i)A455E [38-40] (labeled B in fig. 2); (ii) P574H [40, 41] (labeled S in fig. 2); (iii) A559T [42, 43] (labeled Q in fig. 2).
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ABCC7 p.Pro574His 16132229:131:74
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132 Worth noting is that A455E and P574H are mild alleles, as they have been found associated with preserved pancreatic function and residual secretion of chloride; in addition, a correlation between the A455E mutation and mild pulmonary disease has also been reported [38].
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ABCC7 p.Pro574His 16132229:132:31
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336 333: 95-99 39 De Braekeleer M., Allard C., Leblanc J. P., Simard F. and Aubin G. (1997) Genotype-phenotype correlation in cystic fibrosis patients compound heterozygous for the A455E mutation. Hum. Genet. 101: 208-211 40 Van Oene M., Lukacs G. L. and Rommens J. M. (2000) Cystic fibrosis mutations lead to carboxyl-terminal fragments that highlight an early biogenesis step of the cystic fibrosis transmembrane conductance regulator. J. Biol. Chem. 275: 19577-19584 41 Ostedgaard L. S., Zeiher B. and Welsh M. J. (1999) Processing of CFTR bearing the P574H mutation differs from wild-type and deltaF508-CFTR.
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ABCC7 p.Pro574His 16132229:336:551
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PMID: 16182665 [PubMed] Sarles J et al: "Combining immunoreactive trypsinogen and pancreatitis-associated protein assays, a method of newborn screening for cystic fibrosis that avoids DNA analysis."
No. Sentence Comment
44 A closer look at the results revealed that among the newborns with CF with moderately elevated IRT (50 to <100 ng/mL), 10 had genuine forms of the disease, their genotypes being DF508/DF508 (n = 6), DF508/P574H (n = 1), DF508/G542X (n = 1), DF508/G149R (n = 1), or DF508/?
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ABCC7 p.Pro574His 16182665:44:205
status: NEW
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PMID: 16472140 [PubMed] Becq F et al: "On the discovery and development of CFTR chloride channel activators."
No. Sentence Comment
52 Class V mutations (e.g. A455E, P574H) produce functional proteins with normal Cl-channel activity and regulation but at reduced rate of synthesis [19].
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ABCC7 p.Pro574His 16472140:52:31
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175 Importantly, NS004 is a modulator of several mutated forms of CFTR; P574H [60], K1250A [61], delF508 [31, 35, 61] and G551D [59].
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ABCC7 p.Pro574His 16472140:175:68
status: NEW
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PMID: 16648884 [PubMed] Mishra A et al: "The relevance of sweat testing for the diagnosis of cystic fibrosis in the genomic era."
No. Sentence Comment
100 Some mutations, such as P574H have a defect in folding that is less severe than ΔF508, and as a result the protein reaches the plasma membrane and retains some function.74 Figure 2.
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ABCC7 p.Pro574His 16648884:100:24
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440 Processing of CFTR bearing the P574H mutation differs from wild-type and deltaF508-CFTR.
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ABCC7 p.Pro574His 16648884:440:31
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PMID: 16787252 [PubMed] Verkman AS et al: "CFTR chloride channel drug discovery--inhibitors as antidiarrheals and activators for therapy of cystic fibrosis."
No. Sentence Comment
269 epithelia, but did not correct a different temperature-sensitive CFTR mutant (P574H-CFTR) or a dopamine receptor mutant.
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ABCC7 p.Pro574His 16787252:269:78
status: NEW
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PMID: 18047735 [PubMed] Turnbull EL et al: "The role of the UPS in cystic fibrosis."
No. Sentence Comment
141 Promisingly, the activities of 'Corr` correctors are specific for ΔF508 CFTR in HBE cells and did not affect the CFTR mutants P574H or N1303K, or the dopamine receptor mutant [82].
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ABCC7 p.Pro574His 18047735:141:132
status: NEW
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PMID: 18366345 [PubMed] Caci E et al: "Evidence for direct CFTR inhibition by CFTR(inh)-172 based on Arg347 mutagenesis."
No. Sentence Comment
24 However, NBD mutants such as G551D, G1349D or P574H, do not show an altered sensitivity to CFTRinh-172 [9,12].
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ABCC7 p.Pro574His 18366345:24:46
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PMID: 18493878 [PubMed] Paranjape SM et al: "Atypical cystic fibrosis and CFTR-related diseases."
No. Sentence Comment
64 Determination of the transepithelial nasal potential difference has been beneficial in establishing a CF Table 1 Mutations, sites, and molecular consequences associated with either an atypical presentation of CF respiratory disease or pancreatic sufficiency or late-onset pancreatic insufficiency (http:// www.genet.sickkids.on.ca) Mutation Site Consequence Atypical presentation M1210I Exon 19 Met to Ile at 1210 S1455X Exon 24 Ser to Stop at 1455 1811+18G→A Intron 11 mRNA splicing defect L346P Exon 7 Leu to Pro at 346 Y161D Exon 4 Tyr to Asp at 161 R31C Exon 2 Arg to Cys at 31 I752S Exon 13 Ile to Ser at 752 2811G/T Exon 15 Sequence variation Pancreatic sufficiency or late-onset pancreatic insufficiency R600G Exon 13 Arg to Gly at 600 D1152H Exon 18 Asp to His at 1152 Y89C Exon 3 Tyr to Cys at 89 R117H Exon 4 Arg to His at 117 D110E Exon 4 Asp to Glu at 110 296 + 3insT Intron 2 mRNA splicing defect E217G Exon 6a Glu to Gly at 217 V392G Exon 8 Val to Gly at 392 N1088D Exon 17b Asn to Asp at 1088 S737F Exon 13 Missense 1716+1G→A Intron 10 mRNA splicing defect R334W Exon 7 Arg to Trp at 334 R347P Exon 7 Arg to Pro at 347 A455E Exon 9 Ala to Glu at 455 P574H Exon 12 Pro to His at 574 3850-3T→G Intron 19 mRNA splicing defect diagnosis in many atypical cases.
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ABCC7 p.Pro574His 18493878:64:1179
status: NEW
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ABCC7 p.Pro574His 18493878:64:1193
status: NEW
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PMID: 19837660 [PubMed] Chen JH et al: "Direct sensing of intracellular pH by the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel."
No. Sentence Comment
349 In support of this idea, the effects of acidic and alkaline pHi on D572N-CFTR are reminiscent of the enhanced activity of P574H-CFTR, a CF mutant affecting a residue in the Walker B-D-loop region of NBD1 (52).
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ABCC7 p.Pro574His 19837660:349:122
status: NEW
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PMID: 20059485 [PubMed] Dorfman R et al: "Do common in silico tools predict the clinical consequences of amino-acid substitutions in the CFTR gene?"
No. Sentence Comment
64 Mutations in the CFTR gene grouped by clinical category Cystic fibrosis CFTR-related disease No disease T338I D614G L320V V920L L90S M470V H199R S1251N I203M G550R P111A I148T Q1291H R560K L1388Q L183I R170H I1027T S549R D443Y P499A L1414S T908N R668C S549N A455E E1401K Q151K G27E I1234L Y563N R347P C866R S1118C P1290S R75Q A559T V520F P841R M469V E1401G P67L G85E S50Y E1409K R933G G458V G178R Y1032C R248T I980K G85V V392G L973P L137H T351S R334W I444S V938G R792G R560T R555G L1339F D1305E P574H V1240G T1053I D58G G551D L1335P I918M F994C S945L L558S F1337V R810G D1152H G1247R P574S R766M D579G W1098R H949R F200I R352Q L1077P K1351E M244K L206W M1101K D1154G L375F N1303K R1066C E528D D110Y R347H R1070Q A800G P1021S S549K A1364V V392A damaging` (is supposed to affect protein function or structure) and 'probably damaging` (high confidence of affecting protein function or structure).
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ABCC7 p.Pro574His 20059485:64:495
status: NEW
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57 PI prevalence and in silico prediction scores for 13 most frequent missense mutations identified in Canadian CF patients Mutation Total PI Total (PI + PS) PI prevalence Class PANTHER scorea POLYPHENa SIFTa p.R334W 1 9 0.11 CF-PS -7.4419 Possibly damaging 0.01 p.P67L 2 14 0.14 CF-PS -4.1736 Probably damaging 0 p.R347P 2 12 0.17 CF-PS -7.5259 Possibly damaging 0.01 p.R347H 1 5 0.20 CF-PS -6.8327 Possibly damaging 0 p.A455E 8 39 0.21 CF-PS -8.8641 Probably damaging 0 p.L206W 4 19 0.21 CF-PS -8.5817 Possibly damaging 0 p.P574H 4 7 0.57 CF-PI/PSb -8.1252 Probably damaging 0 p.G85E 15 24 0.63 CF-PI/PSb -7.3194 Possibly damaging 0 p.M1101K 22 33 0.67 CF-PI/PSb -5.8849 Probably damaging 0.01 p.R1066C 7 8 0.88 CF-PI -7.7424 Probably damaging 0 p.G551D 56 59 0.95 CF-PI -9.5654 Probably damaging 0 p.N1303K 47 49 0.96 CF-PI -9.7687 Probably damaging 0 p.V520F 7 7 1.00 CF-PI -7.1652 Benign 0 aPANTHER scores range from zero to negative values (maximum -12).
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ABCC7 p.Pro574His 20059485:57:523
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PMID: 20551307 [PubMed] Da Paula AC et al: "Folding and rescue of a cystic fibrosis transmembrane conductance regulator trafficking mutant identified using human-murine chimeric proteins."
No. Sentence Comment
305 The different effects of K584E on CFTR processing and Cl-channel function are reminiscent of A455E and P574H, two CF mutations associated with a milder clinical phenotype (44).
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ABCC7 p.Pro574His 20551307:305:103
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306 Although production of the mature forms of A455E and P574H was reduced and very much delayed, A455E had channel activity similar to and P574H had channel activity greater than that of WT-CFTR (44).
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ABCC7 p.Pro574His 20551307:306:53
status: NEW
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ABCC7 p.Pro574His 20551307:306:136
status: NEW
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PMID: 20932301 [PubMed] Green DM et al: "Mutations that permit residual CFTR function delay acquisition of multiple respiratory pathogens in CF patients."
No. Sentence Comment
74 For Pa, the hazard ratio Table 1 Classification of CFTR alleles Category Mutation Specific mutations Class I Defective Protein Synthesis (nonsense, frameshift, aberrant splicing) 1078delT, 1154 insTC, 1525-2A > G, 1717-1G > A, 1898+1G > A, 2184delA, 2184 insA, 3007delG, 3120+1G > A, 3659delC, 3876delA, 3905insT, 394delTT, 4010del4, 4016insT, 4326delTC, 4374+1G > T, 441delA, 556delA, 621+1G > T, 621-1G > T, 711+1G > T, 875+1G > C, E1104X, E585X, E60X, E822X, G542X, G551D/R553X, Q493X, Q552X, Q814X, R1066C, R1162X, R553X, V520F, W1282X, Y1092X Class II Abnormal Processing and Trafficking A559T, D979A, ΔF508, ΔI507, G480C, G85E, N1303K, S549I, S549N, S549R Class III Defective Channel Regulation/Gating G1244E, G1349D, G551D, G551S, G85E, H199R, I1072T, I48T, L1077P, R560T, S1255P, S549 (R75Q) Class IV Decreased Channel Conductance A800G, D1152H, D1154G, D614G, delM1140, E822K, G314E, G576A, G622D, G85E, H620Q, I1139V, I1234V, L1335P, M1137V, P67L, R117C, R117P, R117H, R334W, R347H, R347P, R347P/ R347H, R792G, S1251N, V232D Class V Reduced Synthesis and/or Trafficking 2789+5G > A, 3120G > A, 3272-26A > G, 3849+10kbC > T, 5T variant, 621+3A > G, 711+3A > G, A445E, A455E, IVS8 poly T, P574H was increased 3 fold for those with 'Minimal` function when compared to those with 'Residual` function.
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ABCC7 p.Pro574His 20932301:74:1209
status: NEW
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PMID: 21285372 [PubMed] Ostedgaard LS et al: "Cystic fibrosis transmembrane conductance regulator with a shortened R domain rescues the intestinal phenotype of CFTR-/- mice."
No. Sentence Comment
271 BMC Genet 7:18-32. 42. Ostedgaard LS, Zeiher B, Welsh MJ (1999) Processing of CFTR bearing the P574H mutation differs from WT and deltaF508-CFTR.
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ABCC7 p.Pro574His 21285372:271:95
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PMID: 21474639 [PubMed] Rohlfs EM et al: "Cystic fibrosis carrier testing in an ethnically diverse US population."
No. Sentence Comment
131 Four mutations (p.S1255X, p.G330X, c.313delA, p.S364P) were identified only in African Americans, 8 mutations (p.G178R, p.T338I, c.262_ 263delTT, p.M1101K, c.442delA, p.K710X, p.P574H, p.Q1238X)wereidentifiedonlyinCaucasians,and3mu- tations (c.580-1GϾT, c.531delT, p.Q890X) were identified only in Hispanics.
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ABCC7 p.Pro574His 21474639:131:178
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PMID: 21594800 [PubMed] Cai Z et al: "Application of high-resolution single-channel recording to functional studies of cystic fibrosis mutants."
No. Sentence Comment
305 Using biochemical (N) and functional Table 27.1 Comparison of predicted apical membrane Cl- current and measured cAMP-activated apical membrane Cl- current for wild-type and mutant CFTRs CFTR N (%) i (%) Po (%) N × i × Po (%) ICFTR (apical) (%) Wild-type 100 100 100 100 100 F508del 4 100 30 1.2 0 G551D 100 100 2.5 2.5 1.5 R117H 100 86 28 24 15 P574H 15 100 139 21.1 17 N, the number of Cl-channels in the apical membrane; i, single-channel current amplitude; Po, open probability; N × i × Po, the predicted apical membrane Cl- current; ICFTR (apical), measured cAMP-activated apical membrane Cl- current.
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ABCC7 p.Pro574His 21594800:305:359
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312 Table 27.1 compares the predicted values of N × i × Po with the observed values of ICFTR(apical) measured in FRT epithelia for F508del-CFTR and the CF mutants, R117H, G551D and P574H, which disrupt CFTR function by different mechanisms (33, 37, 59, 64, 65).
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ABCC7 p.Pro574His 21594800:312:187
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PMID: 21658632 [PubMed] Becq F et al: "Pharmacological therapy for cystic fibrosis: from bench to bedside."
No. Sentence Comment
56 Interestingly, Zielenski and Tsui [10] assigned A455E and P574H, two CF mutations associated with a milder clinical phenotype (pancreatic sufficiency) to Class V.
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ABCC7 p.Pro574His 21658632:56:58
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58 Moreover, on reaching the cell surface A455E forms Cl-channels with open probability (Po), a measure of channel activity, similar to that of wild-type CFTR, whereas the Po of P574H exceeds that of wild-type CFTR [15].
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ABCC7 p.Pro574His 21658632:58:175
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PMID: 21658649 [PubMed] Bombieri C et al: "Recommendations for the classification of diseases as CFTR-related disorders."
No. Sentence Comment
323 Using biochemical (N) and functional (i and Po) data, the apical CFTR Cl- current generated by the CF-PI mutant p.F508del-CFTR and the CF-PS mutants p.R117H-, p.R334W-, p.R347P-, p.A455E- and p.P574H-CFTR were predicted [166,167].
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ABCC7 p.Pro574His 21658649:323:194
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332 For example, the CF-PS mutant p.P574H-CFTR disrupts CFTR processing, albeit not as severely as p.F508del-CFTR, but generates a CFTR Cl-channel with a Po value greater than that of wild-type CFTR [167].
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ABCC7 p.Pro574His 21658649:332:32
status: NEW
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PMID: 9630075 [PubMed] Arduino C et al: "Congenital bilateral absence of vas deferens with a new missense mutation (P499A) in the CFTR gene."
No. Sentence Comment
52 Other missense mutations in this domain (A455E, P574H, G576A, S549N) have been found associated with a mild CF phenotype and their functional analysis in transfected cells revealed a low transport efficiency of the chloride channel and a reduced protein expression at the apical cell surface, which can explain the mild clinical phenotype (6).
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ABCC7 p.Pro574His 9630075:52:48
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PMID: 9709387 [PubMed] Wilschanski M et al: "Pathology of pancreatic and intestinal disorders in cystic fibrosis."
No. Sentence Comment
152 A small number of more Table 1 Classification of cystic fibrosis gene mutation as severe, mild or indeterminate with respect to pancreatic function Severe Mild Variable (classes 1, I/ or 111) (classes IV or V) (classes IV or V) AF508 R117H G85E 1148T R334W 2789+5G-*A G480C R347P G551D A455E R560T P574H N1303K 3849+1 Okb C-+T G542X G551S W1282X P5748 621 +1 G-T R352Q 1717-1G-T T3381 556delA Adapted from Ref 20 with permission recently described mutations [G85E and 278+5G-÷AI are less clearly determinant with respect to the pancreatic sufficient and pancreatic insufficient phenotypes.
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ABCC7 p.Pro574His 9709387:152:298
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PMID: 9736778 [PubMed] Vankeerberghen A et al: "Characterization of 19 disease-associated missense mutations in the regulatory domain of the cystic fibrosis transmembrane conductance regulator."
No. Sentence Comment
128 So far, only the P574H mutation, located in NBD1, has been shown to give rise to a protein with higher intrinsic chloride transport properties.
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ABCC7 p.Pro574His 9736778:128:17
status: NEW
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PMID: 9843756 [PubMed] Illek B et al: "Genetic disorders of membrane transport. II. Regulation of CFTR by small molecules including HCO3-."
No. Sentence Comment
103 NS-004 restored near normal channel activity from P574H-CFTR (a mild, trafficking-impaired mutation in NBD1) (3).
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ABCC7 p.Pro574His 9843756:103:50
status: NEW
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PMID: 9922378 [PubMed] Schultz BD et al: "Pharmacology of CFTR chloride channel activity."
No. Sentence Comment
579 A second possibility is that the phosporylation state of the channel obtainedsurements, that NS004 similarly activated an additional CFTR mutant, P574H, while failing to activate the CF- during activation by forskolin is distinct from that after activation of the channel by exogenous PKA/ATP in ancausing R560T or DI507 CFTR mutants (70).
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ABCC7 p.Pro574His 9922378:579:146
status: NEW
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PMID: 1717452 [PubMed] Shyamala V et al: "Structure-function analysis of the histidine permease and comparison with cystic fibrosis mutations."
No. Sentence Comment
159 A few mutations are located in the nucleotide-binding pocket, A455E,G458V, and P574H, and may be tolerated because they may have maintained partial activity, especially the two that are not located insites defined as critical for ATP binding by the HisPanalysis.
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ABCC7 p.Pro574His 1717452:159:79
status: NEW
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PMID: 22658665 [PubMed] Ooi CY et al: "Cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations in pancreatitis."
No. Sentence Comment
855 CFTR mutation Total PI Total PI + PS PIP score CFTR mutation Total PI Total PI + PS PIP score 621+1G>T 96 96 1.00 G542X 74 75 0.99 711+1G>T 36 36 1.00 F508del 1276 1324 0.96 I507del 34 34 1.00 1717-1G>A 20 21 0.95 R553X 24 24 1.00 W1282X 19 20 0.95 Q493X 11 11 1.00 N1303K 45 48 0.94 S489X 11 11 1.00 R1162X 12 13 0.92 1154insTC 10 10 1.00 Y1092X 12 13 0.92 3659delC 9 9 1.00 I148T 10 11 0.91 CFTRdele2 7 7 1.00 V520F 9 10 0.90 4016insT 7 7 1.00 G551D 59 67 0.88 E60X 7 7 1.00 L1077P 5 6 0.83 R560T 7 7 1.00 R1066C 5 6 0.83 R1158X 7 7 1.00 2184insA 9 12 0.75 3905insT 6 6 1.00 2143delT 3 4 0.75 I148T;3199del6 5 5 1.00 1161delC 3 4 0.75 2183AA>G 5 5 1.00 3120+1G>A 3 4 0.75 1898+1G>A 5 5 1.00 S549N 3 4 0.75 2347delG 4 4 1.00 G85E 16 22 0.73 Q1313X 3 3 1.00 R117C 2 3 0.67 Q220X 3 3 1.00 M1101K 19 30 0.63 2184delA 3 3 1.00 P574H 3 5 0.60 1078delT 3 3 1.00 474del13BP 1 2 0.50 L1254X 3 3 1.00 R352Q 1 2 0.50 E585X 3 3 1.00 Q1291H 1 2 0.50 3876delA 2 2 1.00 A455E 18 37 0.49 S4X 2 2 1.00 R347P 6 15 0.40 R1070Q 2 2 1.00 2789+5G>A 6 16 0.38 F508C 2 2 1.00 L206W 6 18 0.33 DELI507 2 2 1.00 IVS8-5T 4 16 0.25 Q1411X 2 2 1.00 3272-26A>G 1 4 0.25 365-366insT 2 2 1.00 R334W 1 10 0.10 R709X 2 2 1.00 3849+10kbC>T 2 22 0.09 1138insG 2 2 1.00 P67L 1 14 0.07 CFTRdele2-4 2 2 1.00 R117H 1 25 0.04 3007delG 2 2 1.00 R347H 0 5 0.00 Q814X 2 2 1.00 G178R 0 3 0.00 394delTT 2 2 1.00 E116K 0 2 0.00 406-1G>A 2 2 1.00 875+1G>C 0 2 0.00 R75X 2 2 1.00 V232D 0 2 0.00 CFTRdel2-3 2 2 1.00 D579G 0 2 0.00 E193X 2 2 1.00 L1335P 0 2 0.00 185+1G>T 2 2 1.00 Mild mutations (based on PIP scores) are shaded in gray.
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ABCC7 p.Pro574His 22658665:855:824
status: NEW
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PMID: 22210114 [PubMed] Dong Q et al: "Human-mouse cystic fibrosis transmembrane conductance regulator (CFTR) chimeras identify regions that partially rescue CFTR-DeltaF508 processing and alter its gating defect."
No. Sentence Comment
281 Protein Sci 19: 1932-1947. 35. Ostedgaard LS, Zeiher B, Welsh MJ (1999) Processing of CFTR bearing the P574H mutation differs from wild-type and ΔF508-CFTR.
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ABCC7 p.Pro574His 22210114:281:103
status: NEW
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PMID: 16049310 [PubMed] Schrijver I et al: "Genotyping microarray for the detection of more than 200 CFTR mutations in ethnically diverse populations."
No. Sentence Comment
51 Complete List of Mutations Detectable with the CF APEX Assay CFTR location Amino acid change Nucleotide change 1 E 1 Frameshift 175delC 2 E 2,3 Frameshift del E2, E3 3 E 2 W19C 189 GϾT 4 E 2 Q39X 247 CϾT 5 IVS 2 Possible splicing defect 296 ϩ 12 TϾC 6 E 3 Frameshift 359insT 7 E 3 Frameshift 394delTT 8 E 3 W57X (TAG) 302GϾA 9 E 3 W57X (TGA) 303GϾA 10 E 3 E60X 310GϾT 11 E 3 P67L 332CϾT 12 E 3 R74Q 353GϾA 13 E 3 R75X 355CϾT 14 E 3 G85E 386GϾA 15 E 3 G91R 403GϾA 16 IVS 3 Splicing defect 405 ϩ 1GϾA 17 IVS 3 Possible splicing defect 405 ϩ 3AϾC 18 IVS 3 Splicing defect 406 - 1GϾA 19 E 4 E92X 406GϾT 20 E 4 E92K 406GϾA 21 E 4 Q98R 425AϾG 22 E 4 Q98P 425AϾC 23 E 4 Frameshift 444delA 24 E 4 Frameshift 457TATϾG 25 E 4 R117C 481CϾT 26 E 4 R117H 482GϾA 27 E 4 R117P 482GϾC 28 E 4 R117L 482GϾT 29 E 4 Y122X 498TϾA 30 E 4 Frameshift 574delA 31 E 4 I148T 575TϾC 32 E 4 Splicing defect 621GϾA 33 IVS 4 Splicing defect 621 ϩ 1GϾT 34 IVS 4 Splicing defect 621 ϩ 3AϾG 35 E 5 Frameshift 624delT 36 E 5 Frameshift 663delT 37 E 5 G178R 664GϾA 38 E 5 Q179K 667CϾA 39 IVS 5 Splicing defect 711 ϩ 1GϾT 40 IVS 5 Splicing defect 711 ϩ 1GϾA 41 IVS 5 Splicing defect 712 - 1GϾT 42 E 6a H199Y 727CϾT 43 E 6a P205S 745CϾT 44 E 6a L206W 749TϾG 45 E 6a Q220X 790CϾT 46 E 6b Frameshift 935delA 47 E 6b Frameshift 936delTA 48 E 6b N287Y 991AϾT 49 IVS 6b Splicing defect 1002 - 3TϾG 50 E 7 ⌬F311 3-bp del between nucleotides 1059 and 1069 51 E 7 Frameshift 1078delT 52 E 7 Frameshift 1119delA 53 E 7 G330X 1120GϾT 54 E 7 R334W 1132CϾT 55 E 7 I336K 1139TϾA 56 E 7 T338I 1145CϾT 57 E 7 Frameshift 1154insTC 58 E 7 Frameshift 1161delC 59 E 7 L346P 1169TϾC 60 E 7 R347H 1172GϾA 61 E 7 R347P 1172GϾC 62 E 7 R347L 1172GϾT 63 E 7 R352Q 1187GϾA 64 E 7 Q359K/T360K 1207CϾA and 1211CϾA 65 E 7 S364P 1222TϾC 66 E 8 Frameshift 1259insA 67 E 8 W401X (TAG) 1334GϾA 68 E 8 W401X (TGA) 1335GϾA 69 IVS 8 Splicing changes 1342 - 6 poly(T) variants 5T/7T/9T 70 IVS 8 Splicing defect 1342 - 2AϾC Table 1. Continued CFTR location Amino acid change Nucleotide change 71 E 9 A455E 1496CϾA 72 E 9 Frameshift 1504delG 73 E 10 G480C 1570GϾT 74 E 10 Q493X 1609CϾT 75 E 10 Frameshift 1609delCA 76 E 10 ⌬I507 3-bp del between nucleotides 1648 and 1653 77 E 10 ⌬F508 3-bp del between nucleotides 1652 and 1655 78 E 10 Frameshift 1677delTA 79 E 10 V520F 1690GϾT 80 E 10 C524X 1704CϾA 81 IVS 10 Possible splicing defect 1717 - 8GϾA 82 IVS 10 Splicing defect 1717 - 1GϾA 83 E 11 G542X 1756GϾT 84 E 11 G551D 1784GϾA 85 E 11 Frameshift 1784delG 86 E 11 S549R (AϾC) 1777AϾC 87 E 11 S549I 1778GϾT 88 E 11 S549N 1778GϾA 89 E 11 S549R (TϾG) 1779TϾG 90 E 11 Q552X 1786CϾT 91 E 11 R553X 1789CϾT 92 E 11 R553G 1789CϾG 93 E 11 R553Q 1790GϾA 94 E 11 L558S 1805TϾC 95 E 11 A559T 1807GϾA 96 E 11 R560T 1811GϾC 97 E 11 R560K 1811GϾA 98 IVS 11 Splicing defect 1811 ϩ 1.6 kb AϾG 99 IVS 11 Splicing defect 1812 - 1GϾA 100 E 12 Y563D 1819TϾG 101 E 12 Y563N 1819TϾA 102 E 12 Frameshift 1833delT 103 E 12 D572N 1846GϾA 104 E 12 P574H 1853CϾA 105 E 12 T582R 1877CϾG 106 E 12 E585X 1885GϾT 107 IVS 12 Splicing defect 1898 ϩ 5GϾT 108 IVS 12 Splicing defect 1898 ϩ 1GϾA 109 IVS 12 Splicing defect 1898 ϩ 1GϾC 110 IVS 12 Splicing defect 1898 ϩ 1GϾT 111 E 13 Frameshift 1924del7 112 E 13 del of 28 amino acids 1949del84 113 E 13 I618T 1985TϾC 114 E 13 Frameshift 2183AAϾG 115 E 13 Frameshift 2043delG 116 E 13 Frameshift 2055del9ϾA 117 E 13 D648V 2075TϾA 118 E 13 Frameshift 2105-2117 del13insAGAA 119 E 13 Frameshift 2108delA 120 E 13 R668C 2134CϾT 121 E 13 Frameshift 2143delT 122 E 13 Frameshift 2176insC 123 E 13 Frameshift 2184delA 124 E 13 Frameshift 2184insA 125 E 13 Q685X 2185CϾT 126 E 13 R709X 2257CϾT 127 E 13 K710X 2260AϾT 128 E 13 Frameshift 2307insA 129 E 13 V754M 2392GϾA 130 E 13 R764X 2422CϾT 131 E 14a W846X 2670GϾA 132 E 14a Frameshift 2734delGinsAT 133 E 14b Frameshift 2766del8 134 IVS 14b Splicing defect 2789 ϩ 5GϾA 135 IVS 14b Splicing defect 2790 - 2AϾG 136 E 15 Q890X 2800CϾT 137 E 15 Frameshift 2869insG 138 E 15 S945L 2966CϾT 139 E 15 Frameshift 2991del32 140 E 16 Splicing defect 3120GϾA interrogation: ACCAACATGTTTTCTTTGATCTTAC 3121-2A3G,T S; 5Ј-ACCAACATGTTTTCTTTGATCTTAC A GTTGTTATTAATTGTGATTGGAGCTATAG-3Ј; CAACAA- TAATTAACACTAACCTCGA 3121-2A3G,T AS.
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ABCC7 p.Pro574His 16049310:51:3509
status: NEW
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150 Primers Generated to Create Synthetic Templates That Serve As Positive Mutation Controls Primer name Sense strand 5Ј 3 3Ј Name Antisense strand 5Ј 3 3Ј 175delC synt F T(15)ATTTTTTTCAGGTGAGAAGGTGGCCA 175delC synt R T(15)ATTTGGAGACAACGCTGGCCTTTTCC W19C synt F T(15)TACCAGACCAATTTTGAGGAAAGGAT W19C synt R T(15)ACAGCTAAAATAAAGAGAGGAGGAAC Q39X synt F T(15)TAAATCCCTTCTGTTGATTCTGCTGA Q39X synt R T(15)AGTATATGTCTGACAATTCCAGGCGC 296 ϩ 12TϾC synt F T(15)CACATTGTTTAGTTGAAGAGAGAAAT 296 ϩ 12TϾC synt R T(15)GCATGAACATACCTTTCCAATTTTTC 359insT synt F T(15)TTTTTTTCTGGAGATTTATGTTCTAT 359insT synt R T(15)AAAAAAACATCGCCGAAGGGCATTAA E60X synt F T(15)TAGCTGGCTTCAAAGAAAAATCCTAA E60X synt R T(15)ATCTATCCCATTCTCTGCAAAAGAAT P67L synt F T(15)TTAAACTCATTAATGCCCTTCGGCGA P67L synt R T(15)AGATTTTTCTTTGAAGCCAGCTCTCT R74Q synt F T(15)AGCGATGTTTTTTCTGGAGATTTATG R74Q synt R T(15)TGAAGGGCATTAATGAGTTTAGGATT R75X synt F T(15)TGATGTTTTTTCTGGAGATTTATGTT R75X synt R T(15)ACCGAAGGGCATTAATGAGTTTAGGA W57X(TAG) synt F T(15)AGGATAGAGAGCTGGCTTCAAAGAAA W57X(TAG) synt R T(15)TATTCTCTGCAAAAGAATAAAAAGTG W57X(TGA) synt F T(15)AGATAGAGAGCTGGCTTCAAAGAAAA W57X(TGA) synt R T(15)TCATTCTCTGCAAAAGAATAAAAAGT G91R synt F T(15)AGGGTAAGGATCTCATTTGTACATTC G91R synt R T(15)TTAAATATAAAAAGATTCCATAGAAC 405 ϩ 1GϾA synt F T(15)ATAAGGATCTCATTTGTACATTCATT 405 ϩ 1GϾA synt R T(15)TCCCTAAATATAAAAAGATTCCATAG 405 ϩ 3AϾC synt F T(15)CAGGATCTCATTTGTACATTCATTAT 405 ϩ 3AϾC synt R T(15)GACCCCTAAATATAAAAAGATTCCAT 406 - 1GϾA synt F T(15)AGAAGTCACCAAAGCAGTACAGCCTC 406 - 1GϾA synt R T(15)TTACAAAAGGGGAAAAACAGAGAAAT E92X synt F T(15)TAAGTCACCAAAGCAGTACAGCCTCT E92X synt R T(15)ACTACAAAAGGGGAAAAACAGAGAAA E92K synt F T(15)AAAGTCACCAAAGCAGTACAGCCTCT E92K synt R T(15)TCTACAAAAGGGGAAAAACAGAGAAA 444delA synt F T(15)GATCATAGCTTCCTATGACCCGGATA 444delA synt R T(15)ATCTTCCCAGTAAGAGAGGCTGTACT 574delA synt F T(15)CTTGGAATGCAGATGAGAATAGCTAT 574delA synt R T(15)AGTGATGAAGGCCAAAAATGGCTGGG 621GϾA synt F T(15)AGTAATACTTCCTTGCACAGGCCCCA 621GϾA synt R T(15)TTTCTTATAAATCAAACTAAACATAG Q98P synt F T(15)CGCCTCTCTTACTGGGAAGAATCATA Q98P synt R T(15)GGTACTGCTTTGGTGACTTCCTACAA 457TATϾG synt F T(15)GGACCCGGATAACAAGGAGGAACGCT 457TATϾG synt R T(15)CGGAAGCTATGATTCTTCCCAGTAAG I148T synt F T(15)CTGGAATGCAGATGAGAATAGCTATG I148T synt R T(15)GTGTGATGAAGGCCAAAAATGGCTGG 624delT synt F T(15)CTTAAAGCTGTCAAGCCGTGTTCTAG 624delT synt R T(15)TAAGTCTAAAAGAAAAATGGAAAGTT 663delT synt F T(15)ATGGACAACTTGTTAGTCTCCTTTCC 663delT synt R T(15)CATACTTATTTTATCTAGAACACGGC G178R synt F T(15)AGACAACTTGTTAGTCTCCTTTCCAA G178R synt R T(15)TAATACTTATTTTATCTAGAACACGG Q179K synt F T(15)AAACTTGTTAGTCTCCTTTCCAACAA Q179K synt R T(15)TTCCAATACTTATTTTATCTAGAACA 711 ϩ 5GϾA synt F T(15)ATACCTATTGATTTAATCTTTTAGGC 711 ϩ 5GϾA synt R T(15)TTATACTTCATCAAATTTGTTCAGGT 712 - 1GϾT synt F T(15)TGGACTTGCATTGGCACATTTCGTGT 712 - 1GϾT synt R T(15)TATGGAAAATAAAAGCACAGCAAAAAC H199Y synt F T(15)TATTTCGTGTGGATCGCTCCTTTGCA H199Y synt R T(15)TATGCCAATGCTAGTCCCTGGAAAATA P205S synt F T(15)TCTTTGCAAGTGGCACTCCTCATGGG P205S synt R T(15)TAAGCGATCCACACGAAATGTGCCAAT L206W synt F T(15)GGCAAGTGGCACTCCTCATGGGGCTA L206W synt R T(15)TCAAGGAGCGATCCACACGAAATGTGC Q220X synt F T(15)TAGGCGTCTGCTTTCTGTGGACTTGG Q220X synt R T(15)TATAACAACTCCCAGATTAGCCCCATG 936delTA synt F T(15)AATCCAATCTGTTAAGGCATACTGCT 936delTA synt R T(15)TGATTTTCAATCATTTCTGAGGTAATC 935delA synt F T(15)GAAATATCCAATCTGTTAAGGCATAC 935delA synt R T(15)TATTTCAATCATTTCTGAGGTAATCAC N287Y synt F T(15)TACTTAAGACAGTAAGTTGTTCCAAT N287Y synt R T(15)TATTCAATCATTTTTTCCATTGCTTCT 1002 - 3TϾG synt F T(15)GAGAACAGAACTGAAACTGACTCGGA 1002 - 3TϾG synt R T(15)TCTAAAAAACAATAACAATAAAATTCA 1154insTC syntwt F T(15)ATCTCATTCTGCATTGTTCTGCGCAT 1154insTC syntwt R T(15)TTGAGATGGTGGTGAATATTTTCCGGA 1154insTC syntmt F T(15)TCTCTCATTCTGCATTGTTCTGCGCAT 1154insTC syntmt R T(15)TAGAGATGGTGGTGAATATTTTCCGGA DF311 mt syntV1 F T(15)CCTTCTTCTCAGGGTTCTTTGTGGTG dF311 mt syntV1 R T(15)GAGAAGAAGGCTGAGCTATTGAAGTATC G330X synt F T(15)TGAATCATCCTCCGGAAAATATTCAC G330X synt R T(15)ATTTGATTAGTGCATAGGGAAGCACA S364P synt F T(15)CCTCTTGGAGCAATAAACAAAATACA S364P synt R T(15)GGTCATACCATGTTTGTACAGCCCAG Q359K/T360K mt synt F T(15)AAAAAATGGTATGACTCTCTTGGAGC Q359K/T360K mt synt R T(15)TTTTTTACAGCCCAGGGAAATTGCCG 1078delT synt F T(15)CTTGTGGTGTTTTTATCTGTGCTTCC 1078delT synt R T(15)CAAGAACCCTGAGAAGAAGAAGGCTG 1119delA synt F T(15)CAAGGAATCATCCTCCGGAAAATATT 1119delA synt R T(15)CTTGATTAGTGCATAGGGAAGCACAG 1161delC synt F T(15)GATTGTTCTGCGCATGGCGGTCACTC 1161delC synt R T(15)TCAGAATGAGATGGTGGTGAATATTT T338I synt F T(15)TCACCATCTCATTCTGCATTGTTCTG T338I synt R T(15)ATGAATATTTTCCGGAGGATGATTCC R352Q synt F T(15)AGCAATTTCCCTGGGCTGTACAAACA R352Q synt R T(15)TGAGTGACCGCCATGCGCAGAACAAT L346P synt F T(15)CGCGCATGGCGGTCACTCGGCAATTT L346P synt R T(15)GGAACAATGCAGAATGAGATGGTGGT 1259insA synt F T(15)AAAAAGCAAGAATATAAGACATTGGA 1259insA synt R T(15)TTTTTGTAAGAAATCCTATTTATAAA W401X(TAG)mtsynt F T(15)AGGAGGAGGTCAGAATTTTTAAAAAA W401X(TAG)mtsynt R T(15)TAGAAGGCTGTTACATTCTCCATCAC W401X(TGA) synt F T(15)AGAGGAGGTCAGAATTTTTAAAAAAT W401X(TGA) synt R T(15)TCAGAAGGCTGTTACATTCTCCATCA 1342 - 2AϾC synt F T(15)CGGGATTTGGGGAATTATTTGAGAAA 1342 - 2AϾC synt R T(15)GGTTAAAAAAACACACACACACACAC 1504delG synt F T(15)TGATCCACTGTAGCAGGCAAGGTAGT 1504delG synt R T(15)TCAGCAACCGCCAACAACTGTCCTCT G480C synt F T(15)TGTAAAATTAAGCACAGTGGAAGAAT G480C synt R T(15)ACTCTGAAGGCTCCAGTTCTCCCATA C524X synt F T(15)ACAACTAGAAGAGGTAAGAAACTATG C524X synt R T(15)TCATGCTTTGATGACGCTTCTGTATC V520F synt F T(15)TTCATCAAAGCAAGCCAACTAGAAGA V520F synt R T(15)AGCTTCTGTATCTATATTCATCATAG 1609delCA synt F T(15)TGTTTTCCTGGATTATGCCTGGCACC 1609delCA synt R T(15)CAGAACAGAATGAAATTCTTCCACTG 1717 - 8GϾA synt F T(15)AGTAATAGGACATCTCCAAGTTTGCA 1717 - 8GϾA synt R T(15)TAAAAATAGAAAATTAGAGAGTCACT 1784delG synt F T(15)AGTCAACGAGCAAGAATTTCTTTAGC 1784delG synt R T(15)ACTCCACTCAGTGTGATTCCACCTTC A559T synt F T(15)ACAAGGTGAATAACTAATTATTGGTC A559T synt R T(15)TTAAAGAAATTCTTGCTCGTTGACCT Q552X synt F T(15)TAACGAGCAAGAATTTCTTTAGCAAG Q552X synt R T(15)AACCTCCACTCAGTGTGATTCCACCT S549R(AϾC) synt F T(15)CGTGGAGGTCAACGAGCAAGAATTTC S549R(AϾC) synt R T(15)GCAGTGTGATTCTACCTTCTCCAAGA S549R(TϾG) synt F T(15)GGGAGGTCAACGAGCAAGTATTTC S549R(TϾG) synt R T(15)CCTCAGTGTGATTCCACCTTCTCCAA L558S synt F T(15)CAGCAAGGTGAATAACTAATTATTGG L558S synt R T(15)GAAGAAATTCTCGCTCGTTGACCTCC 1811 ϩ 1.6 kb AϾG synt F T(15)GTAAGTAAGGTTACTATCAATCACAC 1811 ϩ 1.6 kb AϾG synt R T(15)CATCTCAAGTACATAGGATTCTCTGT 1812 - 1GϾA synt F T(15)AAGCAGTATACAAAGATGCTGATTTG 1812 - 1GϾA synt R T(15)TTAAAAAGAAAATGGAAATTAAATTA D572N synt F T(15)AACTCTCCTTTTGGATACCTAGATGT D572N synt R T(15)TTAATAAATACAAATCAGCATCTTTG P574H synt F T(15)ATTTTGGATACCTAGATGTTTTAACA P574H synt R T(15)TGAGAGTCTAATAAATACAAATCAGC 1833delT synt F T(15)ATTGTATTTATTAGACTCTCCTTTTG 1833delT synt R T(15)CAATCAGCATCTTTGTATACTGCTCT Table 4. Continued Primer name Sense strand 5Ј 3 3Ј Name Antisense strand 5Ј 3 3Ј Y563D synt F T(15)GACAAAGATGCTGATTTGTATTTATT Y563D synt R T(15)CTACTGCTCTAAAAAGAAAATGGAAA T582R synt F T(15)GAGAAAAAGAAATATTTGAAAGGTAT T582R synt R T(15)CTTAAAACATCTAGGTATCCAAAAGG E585X synt F T(15)TAAATATTTGAAAGGTATGTTCTTTG E585X synt R T(15)ATTTTTCTGTTAAAACATCTAGGTAT 1898 ϩ 5GϾT synt F T(15)TTTCTTTGAATACCTTACTTATATTG 1898 ϩ 5GϾT synt R T(15)AATACCTTTCAAATATTTCTTTTTCT 1924del7 synt F T(15)CAGGATTTTGGTCACTTCTAAAATGG 1924del7 synt R T(15)CTGTTAGCCATCAGTTTACAGACACA 2055del9ϾA synt F T(15)ACATGGGATGTGATTCTTTCGACCAA 2055del9ϾA synt R T(15)TCTAAAGTCTGGCTGTAGATTTTGGA D648V synt F T(15)TTTCTTTCGACCAATTTAGTGCAGAA D648V synt R T(15)ACACATCCCATGAGTTTTGAGCTAAA K710X synt F T(15)TAATTTTCCATTGTGCAAAAGACTCC K710X synt R T(15)ATCGTATAGAGTTGATTGGATTGAGA I618T synt F T(15)CTTTGCATGAAGGTAGCAGCTATTTT I618T synt R T(15)GTTAATATTTTGTCAGCTTTCTTTAA R764X synt F T(15)TGAAGGAGGCAGTCTGTCCTGAACCT R764X synt R T(15)ATGCCTGAAGCGTGGGGCCAGTGCTG Q685X synt F T(15)TAATCTTTTAAACAGACTGGAGAGTT Q685X synt R T(15)ATTTTTTTGTTTCTGTCCAGGAGACA R709X synt F T(15)TGAAAATTTTCCATTGTGCAAAAGAC R709X synt R T(15)ATATAGAGTTGATTGGATTGAGAATA V754M synt F T(15)ATGATCAGCACTGGCCCCACGCTTCA V754M synt R T(15)TGCTGATGCGAGGCAGTATCGCCTCT 1949del84 synt F T(15)AAAAATCTACAGCCAGACTTTATCTC 1949del84 synt R T(15)TTTTTAGAAGTGACCAAAATCCTAGT 2108delA synt F T(15)GAATTCAATCCTAACTGAGACCTTAC 2108delA synt R T(15)ATTCTTCTTTCTGCACTAAATTGGTC 2176insC synt F T(15)CCAAAAAAACAATCTTTTAAACAGACTGGAGAG 2176insC synt R T(15)GGTTTCTGTCCAGGAGACAGGAGCAT 2184delA synt F T(15)CAAAAAACAATCTTTTAAACAGACTGG 2184delA synt R T(15)GTTTTTTGTTTCTGTCCAGGAGACAG 2105-2117 del13 synt F T(15)AAACTGAGACCTTACACCGTTTCTCA 2105-2117 del13 synt R T(15)TTTCTTTCTGCACTAAATTGGTCGAA 2307insA synt F T(15)AAAGAGGATTCTGATGAGCCTTTAGA 2307insA synt R T(15)TTTCGATGCCATTCATTTGTAAGGGA W846X synt F T(15)AAACACATACCTTCGATATATTACTGTCCAC W846X synt R T(15)TCATGTAGTCACTGCTGGTATGCTCT 2734G/AT synt F T(15)TTAATTTTTCTGGCAGAGGTAAGAAT 2734G/AT synt R T(15)TTAAGCACCAAATTAGCACAAAAATT 2766del8 synt F T(15)GGTGGCTCCTTGGAAAGTGAGTATTC 2766del8 synt R T(15)CACCAAAGAAGCAGCCACCTGGAATGG 2790 - 2AϾG synt F T(15)GGCACTCCTCTTCAAGACAAAGGGAA 2790 - 2AϾG synt R T(15)CGTAAAGCAAATAGGAAATCGTTAAT 2991del32 synt F T(15)TTCAACACGTCGAAAGCAGGTACTTT 2991del32 synt R T(15)AAACATTTTGTGGTGTAAAATTTTCG Q890X synt F T(15)TAAGACAAAGGGAATAGTACTCATAG Q890X synt R T(15)AAAGAGGAGTGCTGTAAAGCAAATAG 2869insG synt F T(15)GATTATGTGTTTTACATTTACGTGGG 2869insG synt R T(15)CACGAACTGGTGCTGGTGATAATCAC 3120GϾA synt F T(15)AGTATGTAAAAATAAGTACCGTTAAG 3120GϾA synt R T(15)TTGGATGAAGTCAAATATGGTAAGAG 3121 - 2AϾT synt F T(15)TGTTGTTATTAATTGTGATTGGAGCT 3121 - 2AϾT synt R T(15)AGTAAGATCAAAGAAAACATGTTGGT 3132delTG synt F T(15)TTGATTGGAGCCATAGCAGTTGTCGC 3132delTG synt R T(15)AATTAATAACAACTGTAAGATCAAAG 3271delGG synt F T(15)ATATGACAGTGAATGTGCGATACTCA 3271delGG synt R T(15)ATTCAGATTCCAGTTGTTTGAGTTGC 3171delC synt F T(15)ACCTACATCTTTGTTGCAACAGTGCC 3171delC synt R T(15)AGGTTGTAAAACTGCGACAACTGCTA 3171insC synt F T(15)CCCCTACATCTTTGTTGCTACAGTGC 3171insC synt R T(15)GGGGTTGTAAAACTGCGACAACTGCT 3199del6 synt F T(15)GAGTGGCTTTTATTATGTTGAGAGCATAT 3199del6 synt R T(15)CCACTGGCACTGTTGCAACAAAGATG M1101K synt F T(15)AGAGAATAGAAATGATTTTTGTCATC M1101K synt R T(15)TTTTGGAACCAGCGCAGTGTTGACAG G1061R synt F T(15)CGACTATGGACACTTCGTGCCTTCGG G1061R synt R T(15)GTTTTAAGCTTGTAACAAGATGAGTG R1066L synt F T(15)TTGCCTTCGGACGGCAGCCTTACTTT R1066L synt R T(15)AGAAGTGTCCATAGTCCTTTTAAGCT R1070P synt F T(15)CGCAGCCTTACTTTGAAACTCTGTTC R1070P synt R T(15)GGTCCGAAGGCACGAAGTGTCCATAG L1077P synt F T(15)CGTTCCACAAAGCTCTGAATTTACAT L1077P synt R T(15)GGAGTTTCAAAGTAAGGCTGCCGTCC W1089X synt F T(15)AGTTCTTGTACCTGTCAACACTGCGC W1089X synt R T(15)TAGTTGGCAGTATGTAAATTCAGAGC L1093P synt F T(15)CGTCAACACTGCGCTGGTTCCAAATG L1093P synt R T(15)GGGTACAAGAACCAGTTGGCAGTATG W1098R synt F T(15)CGGTTCCAAATGAGAATAGAAATGAT W1098R synt R T(15)GGCGCAGTGTTGACAGGTACAAGAAC Q1100P synt F T(15)CAATGAGAATAGAAATGATTTTTGTC Q1100P synt R T(15)GGGAACCAGCGCAGTGTTGACAGGTA D1152H synt F T(15)CATGTGGATAGCTTGGTAAGTCTTAT D1152H synt R T(15)GTATGCTGGAGTTTACAGCCCACTGC R1158X synt F T(15)TGATCTGTGAGCCGAGTCTTTAAGTT R1158X synt R T(15)ACATCTGAAATAAAAATAACAACATT S1196X synt F T(15)GACACGTGAAGAAAGATGACATCTGG S1196X synt R T(15)CAATTCTCAATAATCATAACTTTCGA 3732delA synt F T(15)GGAGATGACATCTGGCCCTCAGGGGG 3732delA synt R T(15)CTCCTTCACGTGTGAATTCTCAATAA 3791delC synt F T(15)AAGAAGGTGGAAATGCCATATTAGAG 3791delC synt R T(15)TTGTATTTTGCTGTGAGATCTTTGAC 3821delT synt F T(15)ATTCCTTCTCAATAAGTCCTGGCCAG 3821delT synt R T(15)GAATGTTCTCTAATATGGCATTTCCA Q1238X synt F T(15)TAGAGGGTGAGATTTGAACACTGCTT Q1238X synt R T(15)AGCCAGGACTTATTGAGAAGGAAATG S1255X (ex19)synt F T(15)GTCTGGCCCTCAGGGGGCCAAATGAC S1255X (ex19) synt R T(15)CGTCATCTTTCTTCACGTGTGAATTC S1255X;L synt F T(15)AAGCTTTTTTGAGACTACTGAACACT S1255X;L synt R T(15)TATAACAAAGTAATCTTCCCTGATCC 3849 ϩ 4AϾG synt F T(15)GGATTTGAACACTGCTTGCTTTGTTA 3849 ϩ 4AϾG synt R T(15)CCACCCTCTGGCCAGGACTTATTGAG 3850 - 1GϾA synt F T(15)AGTGGGCCTCTTGGGAAGAACTGGAT 3850 - 1GϾA synt R T(15)TTATAAGGTAAAAGTGATGGGATCAC 3905insT synt F T(15)TTTTTTTGAGACTACTGAACACTGAA 3905insT synt R T(15)AAAAAAAGCTGATAACAAAGTACTCT 3876delA synt F T(15)CGGGAAGAGTACTTTGTTATCAGCTT 3876delA synt R T(15)CGATCCAGTTCTTCCCAAGAGGCCCA G1244V synt F T(15)TAAGAACTGGATCAGGGAAGAGTACT G1244V synt R T(15)ACCAAGAGGCCCACCTATAAGGTAAA G1249E synt F T(15)AGAAGAGTACTTTGTTATCAGCTTTT G1249E synt R T(15)TCTGATCCAGTTCTTCCCAAGAGGCC S1251N synt F T(15)ATACTTTGTTATCAGCTTTTTTGAGACTACTG S1251N synt R T(15)TTCTTCCCTGATCCAGTTCTTCCCAA S1252P synt F T(15)CCTTTGTTATCAGCTTTTTTGAGACT S1252P synt R T(15)GACTCTTCCCTGATCCAGTTCTTCCC D1270N synt F T(15)AATGGTGTGTCTTGGGATTCAATAAC D1270N synt R T(15)TGATCTGGATTTCTCCTTCAGTGTTC W1282R synt F T(15)CGGAGGAAAGCCTTTGGAGTGATACC W1282R synt R T(15)GCTGTTGCAAAGTTATTGAATCCCAA R1283K synt F T(15)AGAAAGCCTTTGGAGTGATACCACAG R1283K synt R T(15)TTCCACTGTTGCAAAGTTATTGAATC 4005 ϩ 1GϾA synt F T(15)ATGAGCAAAAGGACTTAGCCAGAAAA 4005 ϩ 1GϾA synt R T(15)TCTGTGGTATCACTCCAAAGGCTTTC 4010del4 synt F T(15)GTATTTTTTCTGGAACATTTAGAAAAAACTTGG 4010del4 synt R T(15)AAAATACTTTCTATAGCAAAAAAGAAAAGAAGAA 4016insT synt F T(15)TTTTTTTCTGGAACATTTAGAAAAAACTTGG 4016insT synt R T(15)AAAAAAATAAATACTTTCTATAGCAAAAAAGAAAAGAAGA CFTRdele21 synt F T(15)TAGGTAAGGCTGCTAACTGAAATGAT CFTRdele21 synt R T(15)CCTATAGCAAAAAAGAAAAGAAGAAGAAAGTATG 4382delA synt F T(15)GAGAGAACAAAGTGCGGCAGTACGAT 4382delA synt R T(15)CTCTATGACCTATGGAAATGGCTGTT Bold, mutation allele of interest; bold and italicized, modified nucleotide.
X
ABCC7 p.Pro574His 16049310:150:6858
status: NEW
X
ABCC7 p.Pro574His 16049310:150:6903
status: NEW
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PMID: 15858154 [PubMed] Schrijver I et al: "Diagnostic testing by CFTR gene mutation analysis in a large group of Hispanics: novel mutations and assessment of a population-specific mutation spectrum."
No. Sentence Comment
94 D572N was identified in a Russian patient22 and P574H was identified in two patients with pancreatic sufficiency.23 The serine residue at position 573 is highly conserved across species, as are at least seven residues on either side in mammals, and two in amphibians and fish.24 A second mutation in this subject was not identified.
X
ABCC7 p.Pro574His 15858154:94:48
status: NEW
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PMID: 12458151 [PubMed] Powell K et al: "Therapeutic approaches to repair defects in deltaF508 CFTR folding and cellular targeting."
No. Sentence Comment
92 The effect the DF508, as well as the N1303K, and P574H.
X
ABCC7 p.Pro574His 12458151:92:49
status: NEW
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93 on band C formation was reversible so that when the Class III mutations are defective in regulation of cells were placed back into culture at 37 8C, the chloride conductance through the CFTR at the amount of fully mature protein decreased with a plasma membrane and include the G551D and half-life of approximately 7 h (similar to wild type) G551S mutations.
X
ABCC7 p.Pro574His 12458151:93:49
status: NEW
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PMID: 12454843 [PubMed] Durno C et al: "Genotype and phenotype correlations in patients with cystic fibrosis and pancreatitis."
No. Sentence Comment
105 CFTR Genotypes Among CF Patients With PS With and Without Pancreatitis Two mutations (n) ⌬F508/R117H (9) ⌬F508/(5T) (6) ⌬F508/3272-26A 3 G (4) ⌬F508/R347H (2) ⌬F508/P574H (2) ⌬F508/875 ϩ 1G Ͼ C (2) ⌬F508/3849 ϩ 10kb C 3 T (1) ⌬F508/A455E (1) ⌬F508/D614G (1) ⌬F508/G85E (1) ⌬F508/R347P (1) ⌬F508/S1251N (1) ⌬F508/⌬F508a (1) ⌬F508/3120G Ͼ A (1) ⌬F508/G551Da (1) G542X/R117H (1) R560T/L206W (1) R117H/R117H (1) R31L/P67L (1) 1461ins4 (AGAT)/G85E (1) G551D/(5T) (1) R1066C/3849 ϩ 10kb C Ͼ T (1) G551D/3849 ϩ 10kb C Ͼ T (1) R334W/R334W (1) R334W/681delC (1) W1282X/3489 ϩ 10kb C Ͼ T (1) One mutation (n) ⌬F508/- (18) L1077P/- (1) W1282X/- (1) M1137V/- (1) G551D/- (1) R347H/- (1) Q30X1/- (1) G1244E/- (1) R117H/- (1) 621 ϩ 2G621 ϩ 1G 3 T/- (1) NOTE.
X
ABCC7 p.Pro574His 12454843:105:200
status: NEW
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PMID: 12079272 [PubMed] Naruse S et al: "Cystic fibrosis and related diseases of the pancreas."
No. Sentence Comment
62 is observed only when normal CFTR function is less than 1%.13 In general, patients with pancreatic insuciency are homozygous or compound heterozygous for two severe mutations (class I, II or III in Figure 3), such as DF508, DI507, Q493X, G542X, R553X, W1282X, 621 ‡ 1G 4 T, 1717-1G 4 A, 556delA, 3659delC, I148T, G480C, V520F, G551D, and R560T, whereas the PS phenotype occurs in patients who have one or two mild CFTR mutations, such as R117H, R334W, R347P, A455E, and P574H (class IV or V).5,20 EXOCRINE PANCREAS IN CYSTIC FIBROSIS Pathology of the pancreas in CF There is a spectrum of pancreatic abnormalities in CF irrespective of age.21,22 Pancreatic lesions may be absent in an individual case, but in long-standing CF the pancreas is small, hard and nodular with increased fat and multiple cysts; hence the name `cystic ®brosis of the pancreas'.
X
ABCC7 p.Pro574His 12079272:62:481
status: NEW
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64 is observed only when normal CFTR function is less than 1%.13 In general, patients with pancreatic insuQciency are homozygous or compound heterozygous for two severe mutations (class I, II or III in Figure 3), such as DF508, DI507, Q493X, G542X, R553X, W1282X, 621 W 1G 4 T, 1717-1G 4 A, 556delA, 3659delC, I148T, G480C, V520F, G551D, and R560T, whereas the PS phenotype occurs in patients who have one or two mild CFTR mutations, such as R117H, R334W, R347P, A455E, and P574H (class IV or V).5,20 EXOCRINE PANCREAS IN CYSTIC FIBROSIS Pathology of the pancreas in CF There is a spectrum of pancreatic abnormalities in CF irrespective of age.21,22 Pancreatic lesions may be absent in an individual case, but in long-standing CF the pancreas is small, hard and nodular with increased fat and multiple cysts; hence the name `cystic &#ae;brosis of the pancreas'.
X
ABCC7 p.Pro574His 12079272:64:479
status: NEW
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PMID: 10692317 [PubMed] Xie J et al: "Conformation, independent of charge, in the R domain affects cystic fibrosis transmembrane conductance regulator channel openings."
No. Sentence Comment
272 Although other CFTR mutants have chloride transport in excess of WT CFTR, they are either not processed efficiently (e.g., P574H or H949Y) (Sheppard et al., 1996a, b; Seibert et al., 1996) or open without PKA stimulation (e.g., CFTR-D836X), and thus are not subject to physiologic regulation (Sheppard et al., 1994).
X
ABCC7 p.Pro574His 10692317:272:123
status: NEW
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274 Although other CFTR mutants have chloride transport in excess of WT CFTR, they are either not processed efficiently (e.g., P574H or H949Y) (Sheppard et al., 1996a, b; Seibert et al., 1996) or open without PKA stimulation (e.g., CFTR-D836X), and thus are not subject to physiologic regulation (Sheppard et al., 1994).
X
ABCC7 p.Pro574His 10692317:274:123
status: NEW
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PMID: 10923036 [PubMed] Claustres M et al: "Spectrum of CFTR mutations in cystic fibrosis and in congenital absence of the vas deferens in France."
No. Sentence Comment
107 f 306insA, W79X, R117C, P205S, L227R, I336K, 1248+1G>A, 1609delCA, 1717-8G>A, S549R(T>G), S549N, 1812-1G>A, P574H, 2176insC, R709X, E827X, D836Y, 3007delG, L1065P, L1077P, H1085R, M1101K, 4021insT.
X
ABCC7 p.Pro574His 10923036:107:108
status: NEW
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141 Some mutants are misprocessed (class II) but generate channels that retain significant activity which is sufficient to confer a milder clinical phenotype (for instance, A455E or P574H).
X
ABCC7 p.Pro574His 10923036:141:178
status: NEW
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171 CFTR Mutation Genotypes Identified Both in Cystic Fibrosis (CF) and in Congenital Bilateral Absence of the Vas Deferens (CBAVD) CF CBAVD F508del/5T 3 143 F508del/2789+5G>A 53 1 F508del/3272-26A>G 17 4 F508del/R117H* 10 39 F508del/R117C 2 2 F508del/L206W 12 4 F508del/R347H 10 5 F508del/R347L 1 1 F508del/D443Y 1 5 F508del/Y569C 1 1 F508del/P574H 3 1 F508del/G628R(G>A) 2 1 F508del/V920M 1 1 F508del/R1070W 2 3 F508del/D1152H 6 8 F508del/S1235R 3 1 F508del/T1246I 1 1 F508del/D1270N+R74W 2 3 F508delN1303I 1 1 3659delC/R347H 1 1 G542X/T338I 2 2 R347H/R1066H 1 1 *The only case with CF whose alleles at IVS8(T)n were reported had mutation R117H associated with a 5T allele.
X
ABCC7 p.Pro574His 10923036:171:340
status: NEW
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PMID: 10518014 [PubMed] Zerhusen B et al: "Function of the second nucleotide-binding fold in the CFTR chloride channel."
No. Sentence Comment
212 Mutations in di¡erent regions of CFTR, either changes in single amino acids, i.e. vF508 and P574H [17], or regional deletions, i.e. vexon5 and v19 [20,28], all lead to di¤culty in proper folding and have a signi'cant impact on the processing of CFTR proteins.
X
ABCC7 p.Pro574His 10518014:212:97
status: NEW
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211 Mutations in di&#a1;erent regions of CFTR, either changes in single amino acids, i.e. vF508 and P574H [17], or regional deletions, i.e. vexon5 and v19 [20,28], all lead to di&#a4;culty in proper folding and have a signi'cant impact on the processing of CFTR proteins.
X
ABCC7 p.Pro574His 10518014:211:96
status: NEW
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PMID: 9674722 [PubMed] Schwiebert EM et al: "Cystic fibrosis: a multiple exocrinopathy caused by dysfunctions in a multifunctional transport protein."
No. Sentence Comment
223 They include another deletion mutation at amino acid position 507 (⌬I507), several missense mutations (F508C, G551D, G551S, A455E, R553Q, P574H, S549N, A559T), and some nonsense mutations (G542X, R553X, Q493X).
X
ABCC7 p.Pro574His 9674722:223:145
status: NEW
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238 Some of these mutations, however, such as A455E, P574H and G551S have been associated either with less severe pulmonary disease and/or less compromised Cl- channel function.
X
ABCC7 p.Pro574His 9674722:238:49
status: NEW
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PMID: 9454574 [PubMed] Wigley WC et al: "Transmembrane domain of cystic fibrosis transmembrane conductance regulator: design, characterization, and secondary structure of synthetic peptides m1-m6."
No. Sentence Comment
261 The critical importance of this intramembrane residue to CFTR structure is highlighted by the CF-causing mutation of proline 205 to serine (54), which prevents proper folding and processing of CFTR (52) and causes a form of cystic fibrosis similar to other misprocessing mutants such as A455E and P574H (55).
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ABCC7 p.Pro574His 9454574:261:297
status: NEW
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PMID: 9511935 [PubMed] Bianchet MA et al: "Modeling of nucleotide binding domains of ABC transporter proteins based on a F1-ATPase/recA topology: structural model of the nucleotide binding domains of the cystic fibrosis transmembrane conductance regulator (CFTR)."
No. Sentence Comment
360 The CFTR NBD1 model that results (Fig. 6) gathers the disease causing mutations in three different clusters: (1) mutations affecting the nucleotide binding pocket and the putative general base: A455E, G458V, E504Q AI507 AF508 P574H; (2) mutations in motif C which are probably related to an interaction with region D: S549[R,N,I] G551[S,D], R553Q; and (3) mutations within or near motif B, L558S, A559T, R560T, Y563N and mutations S492F and G480C.
X
ABCC7 p.Pro574His 9511935:360:226
status: NEW
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PMID: 9068292 [PubMed] Wallis C et al: "Diagnosing cystic fibrosis: blood, sweat, and tears."
No. Sentence Comment
29 Pancreatic insuYciency appears to correlate with diVerent gene mutations at the CFTR locus21 (for example R117H, R334W, R347P, P574H), but to date there has not been a satisfactory correlation between a high chloride conduction (that is a high sweat test result )22 or severe pulmonary disease and genotyping.23 The most surprising finding to emanate from the numerous phenotype-genotype correlation studies that festoon the cystic fibrosis literature, is a new understanding of the wide phenotypic range that an individual, homozygous for a mutation in the CFTR gene, can present.
X
ABCC7 p.Pro574His 9068292:29:127
status: NEW
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PMID: 8910333 [PubMed] Seibert FS et al: "Cytoplasmic loop three of cystic fibrosis transmembrane conductance regulator contributes to regulation of chloride channel activity."
No. Sentence Comment
147 A similar effect was noted previously for the CF-associated NBF1 mutation P574H.
X
ABCC7 p.Pro574His 8910333:147:74
status: NEW
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150 The occurrence of substantial activity for these mutants correlates well with the observation that patients affected by the P574H and H949Y mutations suffer from a less severe form of CF and are pancreatic sufficient (Kerem et al., 1990; Ghanem et al., 1994).
X
ABCC7 p.Pro574His 8910333:150:124
status: NEW
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PMID: 8702904 [PubMed] Cotten JF et al: "Effect of cystic fibrosis-associated mutations in the fourth intracellular loop of cystic fibrosis transmembrane conductance regulator."
No. Sentence Comment
84 In Fig. 2 the processing of ⌬F508 and the milder CF-associated mutant, P574H (10), are provided for reference.
X
ABCC7 p.Pro574His 8702904:84:78
status: NEW
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122 Expression of wild-type, ⌬F508, P574H, and ICL4 mutants.
X
ABCC7 p.Pro574His 8702904:122:39
status: NEW
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222 For example, H1085R is misprocessed like ⌬F508, yet it is reported to occur in a patient with a pancreatic sufficient phenotype.
X
ABCC7 p.Pro574His 8702904:222:128
status: NEW
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223 Conversely, our data with the mutants R1066L, R1066H, and A1067T are similar to that obtained with the "mild" mutants A455E and P574H (10) in that they retained partial processing and function.
X
ABCC7 p.Pro574His 8702904:223:128
status: NEW
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83 In Fig. 2 the processing of DF508 and the milder CF-associated mutant, P574H (10), are provided for reference.
X
ABCC7 p.Pro574His 8702904:83:71
status: NEW
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121 Expression of wild-type, DF508, P574H, and ICL4 mutants.
X
ABCC7 p.Pro574His 8702904:121:32
status: NEW
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PMID: 8663008 [PubMed] Sheppard DN et al: "Contribution of proline residues in the membrane-spanning domains of cystic fibrosis transmembrane conductance regulator to chloride channel function."
No. Sentence Comment
192 In this regard they are similar to the CF mutations A455E and P574H that disrupt processing but generate channels that retain significant activity (23).
X
ABCC7 p.Pro574His 8663008:192:62
status: NEW
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217 The mechanism of dysfunction of P205S resembles that of the nucleotide-binding domain 1 pancreatic sufficiency mutants A455E and P574H, which are misprocessed (23).
X
ABCC7 p.Pro574His 8663008:217:129
status: NEW
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198 In this regard they are similar to the CF mutations A455E and P574H that disrupt processing but generate channels that retain significant activity (23).
X
ABCC7 p.Pro574His 8663008:198:62
status: NEW
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225 The mechanism of dysfunction of P205S resembles that of the nucleotide-binding domain 1 pancreatic sufficiency mutants A455E and P574H, which are misprocessed (23).
X
ABCC7 p.Pro574His 8663008:225:129
status: NEW
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PMID: 9156582 [PubMed] Gray MA et al: "Chloride channels and cystic fibrosis of the pancreas."
No. Sentence Comment
117 The second group involves residues within NBD1 (A455E and P574H) which are located close to the walker A (residues 458-464) and B (residues 568-572) motifs, crucial for ATP binding.
X
ABCC7 p.Pro574His 9156582:117:58
status: NEW
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121 However, these channels were fully functional with normal conductance and normal, or greater than normal, Po (P574H).
X
ABCC7 p.Pro574His 9156582:121:110
status: NEW
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118 The second group involves residues within NBD1 (A455E and P574H) which are located close to the walker A (residues 458-464) and B (residues 568-572) motifs, crucial for ATP binding.
X
ABCC7 p.Pro574His 9156582:118:58
status: NEW
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122 However, these channels were fully functional with normal conductance and normal, or greater than normal, Po (P574H).
X
ABCC7 p.Pro574His 9156582:122:110
status: NEW
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PMID: 7472820 [PubMed] Wilschanski M et al: "Correlation of sweat chloride concentration with classes of the cystic fibrosis transmembrane conductance regulator gene mutations."
No. Sentence Comment
43 Defined mutations (each mutation cited in references 8, 23, and 24; numerals in parentheses indicate number of patients): Nonsense mutations-----class I: Frameshift mutations---class I: Splice site mutations-class I: Missense mutations---class HI: Missense mutations---class IV: Partially defective processing---class V: Alternative spficing-----classV: R1162X (3), Y1092X (3), G542X (21), Q552X (2), Q493X (2), w1282x (2), E1104X (1), R553X (6), E585X (l), (all PI) 3659delC (5), 2184delA (4), 4010de14 (1), 556delA (1), 3002delG (1) 3905insT (1), 4016insT (3), 1154insTC (l), 441delA (1), 2184insA (2), 1078delT (1), 4326delTC (3) (all PI) I717-1G--~A (4), 621+lG--*T (10), 711+IG--~T (3), 875+1G-+C (2), 3120+IG-~A (1) (18 PI, 2 PS) G551D (25), N1303K (7), R560T (8), I148T (1), G85E (3), A559T (1), L1077P (2), T1234V (1), (47 PI, 1 PS) R117H (10), R347H (3), R347P (1), D614G (1), S1251N (2), (all PS) P574H (2), A455E (2), (all PS) 3272-26A-+G (4), 3849+10KbC---~T (2), 3120G-+A (1), (all PS) analysis, we further grouped the patients according to the molecular consequences conferred by the CFTR alleles.
X
ABCC7 p.Pro574His 7472820:43:907
status: NEW
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107 Electrophysiologic studies on the mild mutations A455E and P574H revealed that single-channel conductance was not different from the normal CFFR channel but that less protein reached the membrane.
X
ABCC7 p.Pro574His 7472820:107:59
status: NEW
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PMID: 7539891 [PubMed] Gan KH et al: "A cystic fibrosis mutation associated with mild lung disease."
No. Sentence Comment
29 Since the gene for cystic fibrosis was cloned, there have been several studies on associations between the genotype and the phenotype in cystic fibrosis.5-8 A number of mutations (R117H, R334W, R347P, A455E, and P574H) appear to be associated with pancreatic sufficiency9 and residual transmembrane transport of chloride.10,11 The most common mutation, ⌬F508, is associated with pancreatic insufficiency and severe pulmonary disease.5,6 There is great variation in the severity of lung disease, but until now no mutation associated with mild pulmonary disease has been found.
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ABCC7 p.Pro574His 7539891:29:212
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28 Since the gene for cystic fibrosis was cloned, there have been several studies on associations between the genotype and the phenotype in cystic fibrosis.5-8 A number of mutations (R117H, R334W, R347P , A455E, and P574H) appear to be associated with pancreatic sufficiency9 and residual transmembrane transport of chloride.10,11 The most common mutation, F508, is associated with pancreatic insufficiency and severe pulmonary disease.5,6 There is great variation in the severity of lung disease, but until now no mutation associated with mild pulmonary disease has been found.
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ABCC7 p.Pro574His 7539891:28:213
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PMID: 7540133 [PubMed] Champigny G et al: "A change in gating mode leading to increased intrinsic Cl- channel activity compensates for defective processing in a cystic fibrosis mutant corresponding to a mild form of the disease."
No. Sentence Comment
1 Immunochemical and functional analyses indicate that the rank order of CFTR expression at the cell surface is: wild type CFTR > P574H > > AF508 > > R560T - 0.
X
ABCC7 p.Pro574His 7540133:1:128
status: NEW
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2 Patch-clamp analysis indicates that the open probability of P574H Cl- channels is almost twice as high as that ofthe wild type CFTR-CI- channel.
X
ABCC7 p.Pro574His 7540133:2:60
status: NEW
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3 This increased intrinsic activity of individual P574H CFTR-Cl- channels compensates for the lower number of P574H CFTR-Cl- channels reaching the cell surface, and probably explains the milder form of CF associated with the P574H mutation.
X
ABCC7 p.Pro574His 7540133:3:48
status: NEW
X
ABCC7 p.Pro574His 7540133:3:107
status: NEW
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ABCC7 p.Pro574His 7540133:3:108
status: NEW
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4 NS004, a recently described activator, restores near normal CFTR activity in cells expressing the P574H-CFTR channel.
X
ABCC7 p.Pro574His 7540133:4:98
status: NEW
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5 The P574H mutation modifies the gating mode of the channel with a large increase (.X7) in the mean channel open time.
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ABCC7 p.Pro574His 7540133:5:4
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19 This work analyses the molecular properties of a CFTR protein expressed with a recombinant vaccinia virus, which carries the mutation P574H, previously shown to be associated with a mild form of the disease (Kristidis et al., 1992).
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ABCC7 p.Pro574His 7540133:19:134
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22 Results Immunolocalization of P574H-CFTR and other CFTR mutant proteins Recombinant vaccinia viruses expressing wild type or mutant CFTR were constructed to study the effects of al. P 574 H AF 508 R 560 T Fig. 1.
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ABCC7 p.Pro574His 7540133:22:30
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23 Immunodetection of wild type, AF508-, R560T- and P574H-CFTRs in Vero cells.
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ABCC7 p.Pro574His 7540133:23:49
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24 Standard fluorescence microscopic photomicrographs (A, C, E and G) and XY and XZ confocal optical section (B, D, F and H) of Vero cells expressing wild type (A and B), P574H- (C and D), AF508- (E and F) and R560T-CFTRs (G and H).
X
ABCC7 p.Pro574His 7540133:24:168
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27 A1507, AF508, R560T and to compare them with the P574H mutation.
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ABCC7 p.Pro574His 7540133:27:49
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34 In cells expressing P574H-CFTR, the immunolabelling was mainly intracellular, and preferentially localized in the Golgi region, although some cells also exhibited focal CFTR labelling on the plasma membrane (Figure 1C and D).
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ABCC7 p.Pro574His 7540133:34:20
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38 Figure 2 shows the percentage of cells that were positive for membrane immunolabelling after incubation with the 80 c ._ CD 60 m 40 E c 20 0 _ Vero M Chang I -E _m7A - WT P574H AF508 R560T Fig. 2.
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ABCC7 p.Pro574His 7540133:38:174
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39 Analysis of the cell surface expression of wild type, AF508-, R560T- and P574H-CFTRs.
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ABCC7 p.Pro574His 7540133:39:73
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40 The number of Vero and Chang cells used for the analysis was equal to 125 and 223 for wild type CFTR, 59 and 288 for AF508, 169 and 317 for R560T, 88 and 479 for P574H respectively.
X
ABCC7 p.Pro574His 7540133:40:162
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45 The percentage of cells with positive plasma-membrane labelling was lower for P574H and AF508, but remained significantly higher than zero (20-30% for P574H and 2-6% for AF508).
X
ABCC7 p.Pro574His 7540133:45:78
status: NEW
X
ABCC7 p.Pro574His 7540133:45:151
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46 No R560T-expressing cells exhibited plasma-membrane immunolabelling.
X
ABCC7 p.Pro574His 7540133:46:78
status: NEW
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ABCC7 p.Pro574His 7540133:46:151
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50 A diffuse band of 160-170 kDa corresponding to the fully glycosylated CFTR was only detected in cells infected with the recombinant virus expressing wild type CFTR, but in some experiments small amounts of 160-170 kDa glycosylated protein could be observed in cells expressing AF508 or P574H (not shown).
X
ABCC7 p.Pro574His 7540133:50:286
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52 In addition, the absence of fully glycosylated forms of CFTR in cells expressing A1507, AF508, R560T and P574H mutant proteins confirms that these mutations are associated with defective processing.
X
ABCC7 p.Pro574His 7540133:52:105
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53 Functional characterization of P574H-CFTR as compared with other CFTR mutant proteins The functional activity of CFTR mutant proteins was measured in two different experimental conditions that permit activation of the CFTR-CI- channels.
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ABCC7 p.Pro574His 7540133:53:31
status: NEW
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ABCC7 p.Pro574His 7540133:53:105
status: NEW
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54 In the first 'Mock WT P574H R560TAF508 A1507 170kDam F 140kDamp _ mm ----MD Fig. 3.
X
ABCC7 p.Pro574His 7540133:54:22
status: NEW
X
ABCC7 p.Pro574His 7540133:54:31
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55 Western blotting analysis of the 'mock'-infected, wild type, AF508-, R560T- and P574H-CFTRs.
X
ABCC7 p.Pro574His 7540133:55:22
status: NEW
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ABCC7 p.Pro574His 7540133:55:80
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68 Very different results were observed for P574H (Figure 4C-E).
X
ABCC7 p.Pro574His 7540133:68:41
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71 1050 Istim-Ictrl 700 (pA) 350 P574H R560T A1507 urIC) 0L It) C~ a ° Fig. 4.
X
ABCC7 p.Pro574His 7540133:71:30
status: NEW
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72 Functional analysis of the A1507-, R560T- and P574H-CFTRs.
X
ABCC7 p.Pro574His 7540133:72:30
status: NEW
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ABCC7 p.Pro574His 7540133:72:46
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75 (C) Time-course of the activation of the P574H-CFTR mediated 125p- efflux by 10 ,uM forskolin (O), 20 ,uM NS004 (M) and 10 jM forskolin + 20 ,uM NS004 (A).
X
ABCC7 p.Pro574His 7540133:75:41
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77 Results are shown as mean SD (D) Whole cell patch-clamp studies of wild type, A1507, AF508, R560T and P574H after stimulation by 10 jM forskolin.
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ABCC7 p.Pro574His 7540133:77:102
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78 (E) Activation of the A1507, R560T and P574H -C1- currents (Istim) by 20 jM NS004 and/or forskolin 10 jiM.
X
ABCC7 p.Pro574His 7540133:78:39
status: NEW
X
ABCC7 p.Pro574His 7540133:78:102
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81 wild type, P574H and AF508 respectively).
X
ABCC7 p.Pro574His 7540133:81:11
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84 Values of the Cl- current did not differ very significantly between P574H and the wild type proteins, suggesting that in that configuration there might be no significant difference in activity between the two different channel proteins.
X
ABCC7 p.Pro574His 7540133:84:68
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85 Figure 4E shows that NS004 can stimulate the CFTR-C1- current in P574H-CFTR expressing cells, either when used alone or in combination with forskolin.
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ABCC7 p.Pro574His 7540133:85:65
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86 A Cl- current in R560T and A1507 expressing cells was stimulated by forskolin (Figure 4D), but its amplitude remained always very small in comparison with the Cl- current generated by the expression of wild type CFTR (2.6 and 3.7% of the wild type, for A1507 and R560T respectively), and the stimulating effect of NS004 remained modest (Figure 4E).
X
ABCC7 p.Pro574His 7540133:86:65
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87 The mutant protein P574H has a higher intrinsic Ct channel activity than wild type In order to understand why the P574H mutant protein displayed a high residual C1- channel activity, despite its defective cell-surface expression, the properties of the P574H-Cl- channel were compared with those of the wild type CFTR using the cell-attached patch-clamp technique.
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ABCC7 p.Pro574His 7540133:87:19
status: NEW
X
ABCC7 p.Pro574His 7540133:87:114
status: NEW
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ABCC7 p.Pro574His 7540133:87:252
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88 When cells expressing wild type or P574H Cl- channels were exposed to NS004, an increase in the open probability was noticed in both cases (not shown), but the comparison shown here was performed after stimulation by a 'cocktail' of forskolin (5 ,uM), 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate (cpt-cAMP, 100 ,uM) and isobutylmethylxanthine (IBMX, 100 gM) and in the absence of NS004.
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ABCC7 p.Pro574His 7540133:88:19
status: NEW
X
ABCC7 p.Pro574His 7540133:88:35
status: NEW
X
ABCC7 p.Pro574His 7540133:88:114
status: NEW
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ABCC7 p.Pro574His 7540133:88:251
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89 Long lasting recordings of P574H mutant CFTR and wild type CFTR show the typical difference observed between the activity of the two C1- channels (Figure 5A).
X
ABCC7 p.Pro574His 7540133:89:27
status: NEW
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ABCC7 p.Pro574His 7540133:89:35
status: NEW
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90 Although the unitary current amplitude was not modified by the P574H mutation (Figure 5B), the mean number of active Cl- channels was reduced by a factor of -2 (i.e. 46% of the wild type CFTR) (Figure 5C) and the open probability of the channel was increased by 59% (Figure SD).
X
ABCC7 p.Pro574His 7540133:90:27
status: NEW
X
ABCC7 p.Pro574His 7540133:90:63
status: NEW
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91 The conductance of the P574H C1- channel is 5.1 pS, very similar if not identical to that found for the normal CFTR channel (5.3 pS) under the same conditions.
X
ABCC7 p.Pro574His 7540133:91:23
status: NEW
X
ABCC7 p.Pro574His 7540133:91:63
status: NEW
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92 Figure 5E presents a comparative analysis of the mean open and mean closed times of P574H CFTR and normal CFTR after stimulation by the cocktail of forskolin, cpt-cAMP and IBMX.
X
ABCC7 p.Pro574His 7540133:92:23
status: NEW
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ABCC7 p.Pro574His 7540133:92:84
status: NEW
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93 The P574H mutation led to a large increase (a factor of 6.8) in the mean open time of the Cl- channel to a value of 10.4 s.
X
ABCC7 p.Pro574His 7540133:93:4
status: NEW
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ABCC7 p.Pro574His 7540133:93:84
status: NEW
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97 .-.. 2 4 time(minutes) 6 Istim-lctd (pA) I J * NS004 1OgM U ForskliO±M O NS004 + Forsk 1 E WT P574H N-4.
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ABCC7 p.Pro574His 7540133:97:101
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98 Po-0.72 o.4pAIC _L|j C_. -C ~- - Al - - --- --- -- N-4, Po-0.6 N-8, Po.0.31 cJ/\S71q2 i(pA) 8 4How P574H A _._ E 1.___ *FfoP574H2~~ III , * I I~~~~~~~~- 0 35I~~~~I~~II' InLLLEiJ~~~~+60 mV 60 mV ,.
X
ABCC7 p.Pro574His 7540133:98:99
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99 : wr(rw o P574H (r-7) Mean open Mean doe tl -e Fig. 5.
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ABCC7 p.Pro574His 7540133:99:10
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100 Single channel comparison between wild type and P574H mutant Cl- channels.
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ABCC7 p.Pro574His 7540133:100:48
status: NEW
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ABCC7 p.Pro574His 7540133:100:76
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101 (A) Typical cell-attached recordings on forskolin-stimulated WT-CFTR-infected cell (left) and P574H-CFTR infected cell (right); c = closed state of the channels.
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ABCC7 p.Pro574His 7540133:101:94
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102 (B) Mean I-V relationships for wild type and P574H mutant CFTR.
X
ABCC7 p.Pro574His 7540133:102:10
status: NEW
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ABCC7 p.Pro574His 7540133:102:45
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103 (C) Histograms showing the mean number of active CFTR-Cl- channels in wild type and P574H CFTR expressing cells.
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ABCC7 p.Pro574His 7540133:103:48
status: NEW
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ABCC7 p.Pro574His 7540133:103:84
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104 (D) Histograms showing the mean open probability of active CFTR-Cl- channels in wild type and P574H CFTR expressing cells at ±60 mV.
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ABCC7 p.Pro574His 7540133:104:94
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106 (E) Histograms showing the mean open and closed time for wild type and P574H Cl- channels at -60 mV.
X
ABCC7 p.Pro574His 7540133:106:71
status: NEW
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ABCC7 p.Pro574His 7540133:106:83
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109 No significant Cl- channel activity was observed in cells expressing the two mutant proteins A1507 and R560T-CFTR corresponding to severe mutations.
X
ABCC7 p.Pro574His 7540133:109:71
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117 A different situation was observed with the P574H mutation.
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ABCC7 p.Pro574His 7540133:117:44
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119 In this case the anti-CFTR monoclonal antibody labelled the cell surface of 20-30% of P574H-CFTR expressing cells (Figures 1 and 2).
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ABCC7 p.Pro574His 7540133:119:86
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121 This alteration in the traffic toward the cell surface of the P574H mutant protein was confirmed by functional studies.
X
ABCC7 p.Pro574His 7540133:121:62
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122 The number of active channels per patch (estimated by assuming independence between the channels present in a patch) was about half for P574H as compared with wild type (Figure 5C and D).
X
ABCC7 p.Pro574His 7540133:122:86
status: NEW
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ABCC7 p.Pro574His 7540133:122:136
status: NEW
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123 Immature P574H-CFTR accumulated mainly in the Golgi apparatus and in the cytoplasm, whereas diffuse labelling restricted to the cytoplasm was observed for AF508-CFTR.
X
ABCC7 p.Pro574His 7540133:123:9
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125 An interesting property of the P574H mutant protein is that it is associated with defective sorting and increased intrinsic activity of the Cl- channel (Figures 4 and 5).
X
ABCC7 p.Pro574His 7540133:125:31
status: NEW
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ABCC7 p.Pro574His 7540133:125:136
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126 This increased Cl- channel activity is not due to a change of the channel conductance (5.1 pS and 5.3 pS, for P574H- and WT-CFTR, respectively) but is associated with an increase in the time during which the channels remain open.
X
ABCC7 p.Pro574His 7540133:126:9
status: NEW
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ABCC7 p.Pro574His 7540133:126:110
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128 The open probability of the P574H-CFTR Cl- channel is nearly 60% higher than normal.
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ABCC7 p.Pro574His 7540133:128:28
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129 An initial conclusion from these observations is that mutations associated with mild CF: (i) can lead to partial defective processing, less severe than previously observed for severe mutations, and (ii) are not necessarily associated with a decrease in intrinsic channel activity as previously reported for R 117H, R334W and R347P.
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ABCC7 p.Pro574His 7540133:129:109
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132 A second conclusion is that the 'quality control' which prevents the trafficking of mutated CFTR-Cl- channels to the plasma membrane does not only eliminate mutants with reduced or abolished Cl- channel activity, since the intrinsic activity of the P574H mutant protein is higher than normal.
X
ABCC7 p.Pro574His 7540133:132:249
status: NEW
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134 2421 A E I aL The P574H mutation has been identified in compound heterozygotes where it is associated with other mutations such as AF508 (Kristidis et al., 1992).
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ABCC7 p.Pro574His 7540133:134:20
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136 Results from 125v efflux measurements (Figure 4) indicate that the P574H- Cl- channel activity is -70% of the activity measured with wild type CFTR-expressing Vero cells (as measured by the ratios of the maximal stimulation factors).
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ABCC7 p.Pro574His 7540133:136:67
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137 This would mean, assuming that the activity observed in Vero cells can be extrapolated to polarized epithelial cells, that residual in vivo Cl- transport activity for patients bearing P574H mutation would be at most 35% of the normal value.
X
ABCC7 p.Pro574His 7540133:137:20
status: NEW
X
ABCC7 p.Pro574His 7540133:137:184
status: NEW
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142 The association of the P574H mutation with a mild clinical status of the disease is certainly explained by the relatively high intrinsic activity of the corresponding CFTR-C1- channel.
X
ABCC7 p.Pro574His 7540133:142:23
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150 The P574H mutation also increases the mean closed times at -60 mV but only by a factor of 2.6.
X
ABCC7 p.Pro574His 7540133:150:4
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155 The P574H mutation described in this work favours the long open state situation, and as such might become useful for further mechanistic studies of the Cl- channel activity as have been recently carried out with the normal CFTR protein (Fisher and Machen, 1994).
X
ABCC7 p.Pro574His 7540133:155:4
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164 Light fluorescence microscopy Vero and Chang cells were grown on Lab Tek chamber slides and were infected using a recombinant vaccinia virus expression system carrying either the wild type or the mutant (AF508, R560T or P574H CFTR) cDNAs under the control of the T7 promoter.
X
ABCC7 p.Pro574His 7540133:164:220
status: NEW
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51 A diffuse band of 160-170 kDa corresponding to the fully glycosylated CFTR was only detected in cells infected with the recombinant virus expressing wild type CFTR, but in some experiments small amounts of 160-170 kDa glycosylated protein could be observed in cells expressing AF508 or P574H (not shown).
X
ABCC7 p.Pro574His 7540133:51:286
status: NEW
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56 Western blotting analysis of the 'mock'-infected, wild type, AF508-, R560T- and P574H-CFTRs.
X
ABCC7 p.Pro574His 7540133:56:80
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69 Very different results were observed for P574H (Figure 4C-E).
X
ABCC7 p.Pro574His 7540133:69:41
status: NEW
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73 Functional analysis of the A1507-, R560T- and P574H-CFTRs.
X
ABCC7 p.Pro574His 7540133:73:46
status: NEW
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76 (C) Time-course of the activation of the P574H-CFTR mediated 125p- efflux by 10 ,uM forskolin (O), 20 ,uM NS004 (M) and 10 jM forskolin + 20 ,uM NS004 (A).
X
ABCC7 p.Pro574His 7540133:76:41
status: NEW
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79 (E) Activation of the A1507, R560T and P574H -C1- currents (Istim) by 20 jM NS004 and/or forskolin 10 jiM.
X
ABCC7 p.Pro574His 7540133:79:39
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82 wild type, P574H and AF508 respectively).
X
ABCC7 p.Pro574His 7540133:82:11
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94 The P574H mutation led to a large increase (a factor of 6.8) in the mean open time of the Cl-channel to a value of 10.4 s.
X
ABCC7 p.Pro574His 7540133:94:4
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105 (B) Mean I-V relationships for wild type and P574H mutant CFTR.
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ABCC7 p.Pro574His 7540133:105:45
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107 (D) Histograms showing the mean open probability of active CFTR-Cl-channels in wild type and P574H CFTR expressing cells at &#b1;60 mV.
X
ABCC7 p.Pro574His 7540133:107:93
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120 A different situation was observed with the P574H mutation.
X
ABCC7 p.Pro574His 7540133:120:44
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124 This alteration in the traffic toward the cell surface of the P574H mutant protein was confirmed by functional studies.
X
ABCC7 p.Pro574His 7540133:124:62
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131 The open probability of the P574H-CFTR Cl-channel is nearly 60% higher than normal.
X
ABCC7 p.Pro574His 7540133:131:28
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135 A second conclusion is that the 'quality control' which prevents the trafficking of mutated CFTR-Cl-channels to the plasma membrane does not only eliminate mutants with reduced or abolished Cl-channel activity, since the intrinsic activity of the P574H mutant protein is higher than normal.
X
ABCC7 p.Pro574His 7540133:135:247
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139 Results from 125v efflux measurements (Figure 4) indicate that the P574H- Cl- channel activity is -70% of the activity measured with wild type CFTR-expressing Vero cells (as measured by the ratios of the maximal stimulation factors).
X
ABCC7 p.Pro574His 7540133:139:67
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140 This would mean, assuming that the activity observed in Vero cells can be extrapolated to polarized epithelial cells, that residual in vivo Cl-transport activity for patients bearing P574H mutation would be at most 35% of the normal value.
X
ABCC7 p.Pro574His 7540133:140:183
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145 The association of the P574H mutation with a mild clinical status of the disease is certainly explained by the relatively high intrinsic activity of the corresponding CFTR-C1-channel.
X
ABCC7 p.Pro574His 7540133:145:23
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153 The P574H mutation also increases the mean closed times at -60 mV but only by a factor of 2.6.
X
ABCC7 p.Pro574His 7540133:153:4
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158 The P574H mutation described in this work favours the long open state situation, and as such might become useful for further mechanistic studies of the Cl-channel activity as have been recently carried out with the normal CFTR protein (Fisher and Machen, 1994).
X
ABCC7 p.Pro574His 7540133:158:4
status: NEW
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167 Light fluorescence microscopy Vero and Chang cells were grown on Lab Tek chamber slides and were infected using a recombinant vaccinia virus expression system carrying either the wild type or the mutant (AF508, R560T or P574H CFTR) cDNAs under the control of the T7 promoter.
X
ABCC7 p.Pro574His 7540133:167:220
status: NEW
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PMID: 7543317 [PubMed] Pignatti PF et al: "Increased incidence of cystic fibrosis gene mutations in adults with disseminated bronchiectasis."
No. Sentence Comment
31 List of CFTR gene mutations and DNA polymorphisms screened Mutations R75Q/X/L, G85E, 394deITT 457TAT->G, R117H 621 + 1G->T 711 + 5G->A L206W 875 + 40 A->G 936 del TA 1001 + 11C->T R334W, R347 P/H/L, 1154insTC A455E, V456F DF5O8 1717-IG->A, 1717-8G->A G542X, G551D, Q552X, R553X P574H 1898 + 3A->G 2183 AA->G, 2184delA, R709X D836Y, 2694 T/G 2752-22 A/G 2789 + 5 G->A, 2790-2 A-»G Q890X 3041-71 G/C 3132delTG 3271 + 18 C-»T, 3272-26 A->G H1054D, G1061R, R1066C/H, A1067T, H1085R, Y1092X, 3320 ins5 D1152H R1162X, 3667ins4, 3737delA, 11234V 3849 + 10 kb C-»T, 3850-1 G-»A SI25IN, S1255P, 3905insT, 3898insC, D127ON, W1282X, R1283M, 4002 A/G 4005 + 1 G-»A N1303 K/H, 4029 A/G D1377H Q1411 X 4404 C/T, 4521 G/A Location e 3 e 4 i 4 i 5 e 6a i 6a e 6b i 6b e 7 e 9 e 10 i 10 e 11 e 12 i 12 e 13 e 14a i 14a i 14b e 15 i 15 e 17a i 17a e 17b e 18 e 19 i 19 e 20 i 20 e2l e 22 e 23 e24 Listing is in order of location along the CFTR gene, e = exon; i = intron.
X
ABCC7 p.Pro574His 7543317:31:278
status: NEW
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121 Mutations R75Q/ X/L,621 + 1G->T, 1717-1G->A, P574H,2183AA->G, RI066C, andN13O3K, were examined by restriction site generating PCR as described (30).
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ABCC7 p.Pro574His 7543317:121:45
status: NEW
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PMID: 7534226 [PubMed] Sheppard DN et al: "Mechanism of dysfunction of two nucleotide binding domain mutations in cystic fibrosis transmembrane conductance regulator that are associated with pancreatic sufficiency."
No. Sentence Comment
1 To understand better how mutations in CFTR disrupt Cl- channel function and to learn about the relationship between genotype and phenotype, we studied two CF mutants, A455E and P574H, that are associated with pancreatic sufficiency.
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ABCC7 p.Pro574His 7534226:1:176
status: NEW
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2 A455E and P574H are located close to conserved ATP binding motifs in CFTR.
X
ABCC7 p.Pro574His 7534226:2:10
status: NEW
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5 These mutants had normal or increased Cl- channel activity: A455E had an open-state probability (PO) similar to wild-type, and P574H had an increased PO because bursts of activity were prolonged.
X
ABCC7 p.Pro574His 7534226:5:126
status: NEW
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27 Two such mutations are located within NBD1: A455E and P574H (Kerem et al., 1990; Kristidis et al., 1992).
X
ABCC7 p.Pro574His 7534226:27:54
status: NEW
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29 A455E and P574H affect residues in NBD1 that are located close to the Walker A (residues 458-464) and B (residues 568-572) motifs, respectively.
X
ABCC7 p.Pro574His 7534226:29:10
status: NEW
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31 Therefore, we tested the hypothesis that A455E and P574H might alter the function of CFTR Cl- channels.
X
ABCC7 p.Pro574His 7534226:31:51
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33 86 Oxford University Press Results Expression of A455E and P574H generates cAMP-activated CL currents To learn whether A455E and P574H could form regulated Cl- channels in epithelia, we expressed these mutants in cAMP~ N E ._0. -1 P574H cAMPI A455E 'cAMP 5 pA/cm2L 5 min b Fig. 1. cAMP agonists activate apical membrane CF- currents in FRT epithelia expressing A455E and P574H.
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ABCC7 p.Pro574His 7534226:33:61
status: NEW
X
ABCC7 p.Pro574His 7534226:33:62
status: NEW
X
ABCC7 p.Pro574His 7534226:33:131
status: NEW
X
ABCC7 p.Pro574His 7534226:33:132
status: NEW
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44 Figure 1 shows that A455E and P574H generated cAMP-stimulated apical Cl- currents, but AF508 did not.
X
ABCC7 p.Pro574His 7534226:44:30
status: NEW
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45 However, the magnitude of current generated by A455E and P574H was <20% that of wild-type CFTR in the rank order: wild-type CFTR>> P574H > A455E > AF508.
X
ABCC7 p.Pro574His 7534226:45:30
status: NEW
X
ABCC7 p.Pro574His 7534226:45:57
status: NEW
X
ABCC7 p.Pro574His 7534226:45:131
status: NEW
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46 When we expressed A455E and P574H in HeLa cells and studied them with the whole-cell patch-clamp technique, we found properties qualitatively similar to those observed with wild-type CFTR (Welsh et al., 1992; Riordan, 1993).
X
ABCC7 p.Pro574His 7534226:46:28
status: NEW
X
ABCC7 p.Pro574His 7534226:46:57
status: NEW
X
ABCC7 p.Pro574His 7534226:46:131
status: NEW
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47 Figure 2 shows data from studies of P574H; similar results were obtained with A455E (data not shown).
X
ABCC7 p.Pro574His 7534226:47:28
status: NEW
X
ABCC7 p.Pro574His 7534226:47:36
status: NEW
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51 In contrast, when P574H was expressed using the same conditions, cAMP agonists stimulated current in only five out of 12 cells, IP Cl [CI]E 7 ; - 25~O20 h / -200 Fig. 2.
X
ABCC7 p.Pro574His 7534226:51:18
status: NEW
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52 Whole-cell properties of P574H.
X
ABCC7 p.Pro574His 7534226:52:18
status: NEW
X
ABCC7 p.Pro574His 7534226:52:25
status: NEW
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66 CFTR 0 * I1 -50 * CFTR A A455E * P574H pA Fig. 3.
X
ABCC7 p.Pro574His 7534226:66:33
status: NEW
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67 Single-channel properties of A455E and P574H.
X
ABCC7 p.Pro574His 7534226:67:33
status: NEW
X
ABCC7 p.Pro574His 7534226:67:39
status: NEW
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68 (A) Representative single-channel recordings are from excised inside-out membrane patches from HeLa cells transiently expressing wild-type CFTR, A455E or P574H.
X
ABCC7 p.Pro574His 7534226:68:39
status: NEW
X
ABCC7 p.Pro574His 7534226:68:154
status: NEW
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72 The voltages were -45 (CFTR), -46 (A455E) or -47 mV (P574H).
X
ABCC7 p.Pro574His 7534226:72:53
status: NEW
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73 (B) Single-channel I-V relationships of CFTR (U), A455E (A) and P574H (A).
X
ABCC7 p.Pro574His 7534226:73:53
status: NEW
X
ABCC7 p.Pro574His 7534226:73:64
status: NEW
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76 Conductance was CFT'R 7.76 ± 0.14, A455E 7.40 ± 0.25 and P574H 7.84 ± 0.26 pS; n = 6.
X
ABCC7 p.Pro574His 7534226:76:67
status: NEW
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81 Single-channel properties of A455E and P574H To determine why A455E and P574H generated less macroscopic Cl- current, we examined their single-channel properties in excised, inside-out patches of membrane.
X
ABCC7 p.Pro574His 7534226:81:39
status: NEW
X
ABCC7 p.Pro574His 7534226:81:72
status: NEW
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83 The tracings show that the single-channel current amplitudes of A455E and P574H were similar to that of wild-type CFTR.
X
ABCC7 p.Pro574His 7534226:83:74
status: NEW
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88 However, P574H showed a different pattern of activity.
X
ABCC7 p.Pro574His 7534226:88:9
status: NEW
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92 Figure 3C shows that the PO of A455E was similar to that of wild-type CFTR, whereas the PO of P574H was increased significantly.
X
ABCC7 p.Pro574His 7534226:92:94
status: NEW
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95 We focused primarily on P574H to determine why the P0 of this mutant was greater than that of wild-type CFTR.
X
ABCC7 p.Pro574His 7534226:95:24
status: NEW
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96 Maximum likelihood A 0.5pA 2s 50OC mV 0.6 PO 0.4 0.2- -0.1 - -1.2 0.0 J- - -0.4 A B pi (/S) al (f/S) pi 02I -- C2 _ ° a1 a2 12 CFTR A455E P574H CFTR A455E P574H Fig. 4.
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ABCC7 p.Pro574His 7534226:96:24
status: NEW
X
ABCC7 p.Pro574His 7534226:96:145
status: NEW
X
ABCC7 p.Pro574His 7534226:96:162
status: NEW
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97 Effect of A455E and P574H on single-channel kinetics.
X
ABCC7 p.Pro574His 7534226:97:20
status: NEW
X
ABCC7 p.Pro574His 7534226:97:146
status: NEW
X
ABCC7 p.Pro574His 7534226:97:163
status: NEW
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100 (B) Effect of A455E and P574H on rate constants determined by the maximum likelihood fit to the model in (A).
X
ABCC7 p.Pro574His 7534226:100:24
status: NEW
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101 Data are from four CFTR, two A455E and six P574H single-channel patches.
X
ABCC7 p.Pro574His 7534226:101:24
status: NEW
X
ABCC7 p.Pro574His 7534226:101:43
status: NEW
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103 The voltages were -50 ± 2 (CFTR), -48 ± I (P574H) and -47 ± 1 mV (A455E).
X
ABCC7 p.Pro574His 7534226:103:53
status: NEW
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110 The effect of P574H on the rate constants is summarized in Figure 4B.
X
ABCC7 p.Pro574His 7534226:110:14
status: NEW
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111 The major effect of the P574H mutation was to decrease PI and al to -40% of wild-type values.
X
ABCC7 p.Pro574His 7534226:111:14
status: NEW
X
ABCC7 p.Pro574His 7534226:111:24
status: NEW
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118 All of the changes in rate constants produced by P574H tend to increase Tb.
X
ABCC7 p.Pro574His 7534226:118:49
status: NEW
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119 The increase in burst duration is responsible for the increase in PO (Figure 3C), but it is partially offset by the decreased frequency with which P574H channels move into the bursting state (i.e. the decreased I31).
X
ABCC7 p.Pro574His 7534226:119:49
status: NEW
X
ABCC7 p.Pro574His 7534226:119:147
status: NEW
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120 In two experiments we examined the gating kinetics of single phosphorylated A455E Cl- channels.
X
ABCC7 p.Pro574His 7534226:120:147
status: NEW
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124 Regulation of A455E and P574H by intracellular nucleotides Intracellular MgATP regulates CFTR through interactions with the NBDs.
X
ABCC7 p.Pro574His 7534226:124:24
status: NEW
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127 Therefore, we tested the hypothesis that ATP-dependent regulation of A455E and P574H was altered.
X
ABCC7 p.Pro574His 7534226:127:79
status: NEW
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129 At each concentration of MgATP tested, A455E had PO values similar to those of wild-type CFTR, whereas P574H had higher values of PO.
X
ABCC7 p.Pro574His 7534226:129:103
status: NEW
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131 Figure 5B shows that ADP (1 mM) produced an equivalent inhibition of wild-type CFTR, A455E and P574H.
X
ABCC7 p.Pro574His 7534226:131:95
status: NEW
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133 Analysis of the processing of A455E and P574H The data indicate that the reduced apical membrane Cl- current produced by A455E and P574H cannot be attributed to the reduced function of single mutant channels.
X
ABCC7 p.Pro574His 7534226:133:40
status: NEW
X
ABCC7 p.Pro574His 7534226:133:131
status: NEW
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139 However, band C production was reduced in A455E and P574H and was not apparent until 12-24 h after infection, and only then as a faint A B CFTR A455E P574H MgATP (mM) Fig. 5.
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ABCC7 p.Pro574His 7534226:139:52
status: NEW
X
ABCC7 p.Pro574His 7534226:139:152
status: NEW
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140 Effects of MgATP concentration and ADP on the activity of phosphorylated A455E and P574H channels.
X
ABCC7 p.Pro574His 7534226:140:52
status: NEW
X
ABCC7 p.Pro574His 7534226:140:83
status: NEW
X
ABCC7 p.Pro574His 7534226:140:152
status: NEW
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142 and intracellular MgATP concentration for A455E, P574H and wild-type CFTR Cl- channels.
X
ABCC7 p.Pro574His 7534226:142:49
status: NEW
X
ABCC7 p.Pro574His 7534226:142:77
status: NEW
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143 Data points are the mean ± SEM of n = 4-6 for CFTR (-), n = 5 for A455E (A) and n = 3-5 for P574H (0) at each concentration.
X
ABCC7 p.Pro574His 7534226:143:96
status: NEW
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144 The voltages were -86 ± 1 (CFTR), -49 ± 2 (A455E) and -49 ± 1 mV (P574H).
X
ABCC7 p.Pro574His 7534226:144:78
status: NEW
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147 (B) Effect of intracellular ADP on the activities of CFTR, A455E and P574H Cl- channels.
X
ABCC7 p.Pro574His 7534226:147:69
status: NEW
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149 Values are the mean ± SEM of n = 4 for CFTR, A455E and P574H; the voltages were -49 ± 1 (CFTR), -48 ± 2 (A455E) and -52 ± 1 mV (P574H).
X
ABCC7 p.Pro574His 7534226:149:59
status: NEW
X
ABCC7 p.Pro574His 7534226:149:60
status: NEW
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150 Percent inhibition of NXP0 by 1 mM ADP was CFTR 77 ± 3, A455E 70 ± 3 and P574H 75 ± 6%.
X
ABCC7 p.Pro574His 7534226:150:81
status: NEW
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156 Although the production of mature band C protein was greatly reduced in A455E and P574H, it was greater than that observed with AF508.
X
ABCC7 p.Pro574His 7534226:156:82
status: NEW
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157 Discussion Relationship between apical membrane CL currents and the properties of mutant CFTR Because the A455E and P574H mutations cause CF, we hypothesized that Cl- transport would be reduced in cells expressing these mutants.
X
ABCC7 p.Pro574His 7534226:157:116
status: NEW
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161 If we set each of these variables to 100% for wild-type CFTR, we can then compare the apical Cl- current generated by wild-type and AF508 CFTR with that produced by A455E and P574H. Table I presents values of each variable, the predicted value of fCI(apical) determined by calculating N x i X P0 and the observed value of fCI(apical).
X
ABCC7 p.Pro574His 7534226:161:174
status: NEW
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164 Thus, these data provide a molecular explanation for the quantitative decrease in cAMP-stimulated apical membrane Cl- current generated by A455E and P574H. Table I.
X
ABCC7 p.Pro574His 7534226:164:148
status: NEW
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165 Comparison of predicted apical membrane Cl- current, N X i X PO, and measured cAMP-stimulated apical membrane Cl- current, f'(apical), for wild-type and mutant CFTR N i PO N X i X PO fl'(apical) (%) (%) (%) (%) (%) Wild-type 100 100 100 100 100 AF508 4 100 38 1.6 0 A455E 11 100 93 10.5 8 P574H 15 100 139 21.1 17 N, the number of Cl- channels in the apical membrane; i, single-channel current; PO- open-state probability.
X
ABCC7 p.Pro574His 7534226:165:287
status: NEW
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171 Mechanism of altered gating by A455E and P574H A455 and P574 lie near the conserved Walker A and B motifs of NBD1.
X
ABCC7 p.Pro574His 7534226:171:41
status: NEW
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174 P574H decreased 01, a rate constant that is also controlled by ATP and which describes the transition to the bursting state.
X
ABCC7 p.Pro574His 7534226:174:0
status: NEW
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175 Because of the proximity of P574 to D572 (the Walker B aspartate of NBD1), we speculate that P574H alters the interaction of NBD1 with MgATP, and/ or the consequences thereof, and hence slows the rate of channel opening.
X
ABCC7 p.Pro574His 7534226:175:93
status: NEW
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176 P574H also altered al.
X
ABCC7 p.Pro574His 7534226:176:0
status: NEW
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177 It is interesting to compare the effect that different mutations in NBD1 had on al: P574H decreased a1, A455E did not change al, and K464A (a mutant of the Walker A lysine of NBDl; Carson and Welsh, 1995) appeared to increase the rate of channel closing.
X
ABCC7 p.Pro574His 7534226:177:84
status: NEW
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181 Expression of wild-type CFTR, AF508, A455E and P574H.
X
ABCC7 p.Pro574His 7534226:181:47
status: NEW
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187 The radioactivity in gels of immunoprecipitated and phosphorylated wild-type CFTR, AF508, A455E and P574H was quantitated as described in Materials and methods.
X
ABCC7 p.Pro574His 7534226:187:100
status: NEW
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188 The values are the mean + SEM of n = 3 for CFTR, AF508, A455E and P574H.
X
ABCC7 p.Pro574His 7534226:188:66
status: NEW
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189 understand how CFTR is regulated by the NBDs and to learn how P574H alters this function.
X
ABCC7 p.Pro574His 7534226:189:62
status: NEW
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192 Our data from A455E, plus the finding that P574H altered P2, suggest that the NBDs may also be involved in regulating gating within a burst.
X
ABCC7 p.Pro574His 7534226:192:43
status: NEW
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194 Cellular processing ofA455E and P574H An important mechanism of dysfunction of CFTR containing CF-associated mutations is misprocessing, so that the mutant protein fails to leave the endoplasmic reticulum and traffic to the Golgi complex and then on to the cell surface (Cheng et al., 1990).
X
ABCC7 p.Pro574His 7534226:194:32
status: NEW
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198 Our data indicate that processing mutations are not necessarily an all-or-none defect; in contrast to AF508, some of the mutant A455E and P574H protein had a glycosylation pattern consistent with processing in the Golgi complex and generated cAMP-stimulated apical Cl- currents.
X
ABCC7 p.Pro574His 7534226:198:138
status: NEW
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199 We evaluated the processing of A455E and P574H in HeLa and FRT cells incubated at 37°C.
X
ABCC7 p.Pro574His 7534226:199:41
status: NEW
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204 More strikingly, P574H, which was also misprocessed, formed a channel that was even more active than wild-type.
X
ABCC7 p.Pro574His 7534226:204:17
status: NEW
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209 Implications for cystic fibrosis These data suggest that the mutations A455E and P574H cause CF because they disrupt the processing of CFTR and its delivery to the cell surface.
X
ABCC7 p.Pro574His 7534226:209:81
status: NEW
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210 However, they also suggest that these mutations are associated with a milder (PS) clinical phenotype because a small amount of the 881 CFTR A kDa 210 - 170- 116 - 98 - AF508 N. A455E I I P574H 4- Band C 4- Band B B band C bands A+B P574H aL mutant protein is processed correctly, retains normal or greater than normal Cl- channel function and thus generates residual cAMP-stimulated apical membrane C1- cur- rents.
X
ABCC7 p.Pro574His 7534226:210:187
status: NEW
X
ABCC7 p.Pro574His 7534226:210:232
status: NEW
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220 These values are in the same range as those found for A455E (8%) and P574H (17%).
X
ABCC7 p.Pro574His 7534226:220:69
status: NEW
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223 A455E or P574H could prove to be of value in the development of therapies designed to augment the delivery to and the retention of mutant protein at the plasma membrane (Cheng et al., 1990; Lukacs et al., 1993).
X
ABCC7 p.Pro574His 7534226:223:9
status: NEW
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224 Although AF508 could be used to assess the processing defect, A455E or P574H might prove to be more tractable models for evaluating strategies to correct mislocalization because the processing defect is only partial and the conductance and regulation of the Cl- channels is more nearly normal.
X
ABCC7 p.Pro574His 7534226:224:71
status: NEW
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226 Such strategies might also be applied to patients bearing the A455E and P574H mutations because they appear to have at least some functional protein present in the plasma membrane.
X
ABCC7 p.Pro574His 7534226:226:72
status: NEW
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34 -1 P574H cAMPI A455E 'cAMP 5 pA/cm2L 5 min b Fig. 1. cAMP agonists activate apical membrane CF- currents in FRT epithelia expressing A455E and P574H.
X
ABCC7 p.Pro574His 7534226:34:3
status: NEW
X
ABCC7 p.Pro574His 7534226:34:143
status: NEW
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48 Figure 2 shows data from studies of P574H; similar results were obtained with A455E (data not shown).
X
ABCC7 p.Pro574His 7534226:48:36
status: NEW
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53 Whole-cell properties of P574H.
X
ABCC7 p.Pro574His 7534226:53:25
status: NEW
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69 (A) Representative single-channel recordings are from excised inside-out membrane patches from HeLa cells transiently expressing wild-type CFTR, A455E or P574H.
X
ABCC7 p.Pro574His 7534226:69:154
status: NEW
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74 (B) Single-channel I-V relationships of CFTR (U), A455E (A) and P574H (A).
X
ABCC7 p.Pro574His 7534226:74:64
status: NEW
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77 Conductance was CFT'R 7.76 &#b1; 0.14, A455E 7.40 &#b1; 0.25 and P574H 7.84 &#b1; 0.26 pS; n = 6.
X
ABCC7 p.Pro574His 7534226:77:65
status: NEW
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82 Single-channel properties of A455E and P574H To determine why A455E and P574H generated less macroscopic Cl-current, we examined their single-channel properties in excised, inside-out patches of membrane.
X
ABCC7 p.Pro574His 7534226:82:39
status: NEW
X
ABCC7 p.Pro574His 7534226:82:72
status: NEW
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84 The tracings show that the single-channel current amplitudes of A455E and P574H were similar to that of wild-type CFTR.
X
ABCC7 p.Pro574His 7534226:84:74
status: NEW
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89 However, P574H showed a different pattern of activity.
X
ABCC7 p.Pro574His 7534226:89:9
status: NEW
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93 Figure 3C shows that the PO of A455E was similar to that of wild-type CFTR, whereas the PO of P574H was increased significantly.
X
ABCC7 p.Pro574His 7534226:93:94
status: NEW
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98 Effect of A455E and P574H on single-channel kinetics.
X
ABCC7 p.Pro574His 7534226:98:20
status: NEW
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102 Data are from four CFTR, two A455E and six P574H single-channel patches.
X
ABCC7 p.Pro574His 7534226:102:43
status: NEW
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104 The voltages were -50 &#b1; 2 (CFTR), -48 &#b1; I (P574H) and -47 &#b1; 1 mV (A455E).
X
ABCC7 p.Pro574His 7534226:104:51
status: NEW
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112 The major effect of the P574H mutation was to decrease PI and al to -40% of wild-type values.
X
ABCC7 p.Pro574His 7534226:112:24
status: NEW
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125 Regulation of A455E and P574H by intracellular nucleotides Intracellular MgATP regulates CFTR through interactions with the NBDs.
X
ABCC7 p.Pro574His 7534226:125:24
status: NEW
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128 Therefore, we tested the hypothesis that ATP-dependent regulation of A455E and P574H was altered.
X
ABCC7 p.Pro574His 7534226:128:79
status: NEW
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130 At each concentration of MgATP tested, A455E had PO values similar to those of wild-type CFTR, whereas P574H had higher values of PO.
X
ABCC7 p.Pro574His 7534226:130:103
status: NEW
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132 Figure 5B shows that ADP (1 mM) produced an equivalent inhibition of wild-type CFTR, A455E and P574H.
X
ABCC7 p.Pro574His 7534226:132:95
status: NEW
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134 Analysis of the processing of A455E and P574H The data indicate that the reduced apical membrane Cl-current produced by A455E and P574H cannot be attributed to the reduced function of single mutant channels.
X
ABCC7 p.Pro574His 7534226:134:40
status: NEW
X
ABCC7 p.Pro574His 7534226:134:130
status: NEW
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141 Effects of MgATP concentration and ADP on the activity of phosphorylated A455E and P574H channels.
X
ABCC7 p.Pro574His 7534226:141:83
status: NEW
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PMID: 8825494 [PubMed] Zielenski J et al: "Cystic fibrosis: genotypic and phenotypic variations."
No. Sentence Comment
644 Several missense mutations (e.g. P574H and A455E) are reasoned to have mild molecular consequence because they are found associated with pancreatic sufficiency-a mild phenotype (lOS).
X
ABCC7 p.Pro574His 8825494:644:33
status: NEW
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PMID: 7522998 [PubMed] Tsongalis GJ et al: "Association of pancreatic adenocarcinoma, mild lung disease, and delta F508 mutation in a cystic fibrosis patient."
No. Sentence Comment
44 CorrelatIon of phenotype and genotype of CFTR mutations Key phenotypic Lung disease SweatC1 Exocnne pancreas function Vasdeferens Associated CFTR mutations Pancreatic InsuffIcIent Pancreatic sufficient Normalsweat C1 Severe Less severe Relatively mild Elevated Elevated Normal Insufficient Sufficient Sufficient Absent Absent Absent SF508, G542X, R553X, G5510, Ni 303K, Wi 282X, RI 17H, and others 2789 + 5G>A, R117H, R334W, R347P, A455E, P574H, S945L, G85E, and others G551S, R117H, 3849 + 10kb C>T, and others Congenitalabsence of the vas deferens None Normal or elevated Sufficient Absent F508C, Ri 17H, Di D1152H, and others FIg. 2.
X
ABCC7 p.Pro574His 7522998:44:439
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PMID: 7525963 [PubMed] Chevalier-Porst F et al: "Mutation analysis in 600 French cystic fibrosis patients."
No. Sentence Comment
21 Among the 104 other CFTR mutations tested on the 373 non-AF508 CF chromosomes, none of the following 58 mutations were found: G91R, 435 insA, 444delA, D11OH, 556delA, 557delT, R297Q, 1154insTC, R347L, R352Q, Q359K/T360K, 1221delCT, G480C, Q493R, V520F, C524X, 1706dell7, S549R (A-C), S549N, S549I, G551S, 1784delG, Q552X, L558S, A559T, R560T, R560K, Y563N, P574H, 2307insA, 2522insC, 2556insAT, E827X, Q890X, Y913C, 2991de132 (Dork et al, personal communication), L967S, 3320ins5, 3359delCT, H1085R, R1158X, 3662delA, 3667del4, 3667ins4, 3732delA, 3737delA, W1204X, 3750delAG, I 1234V, Q1238X, 3850- 3T-+G, 3860ins31, S1255X, 3898insC, D1270N, R1283M, F1286S, 4005 + I G-A. Forty-six other mutations were found on at Distribution of CFTR mutations found in our sample ofpopulation (1200 CF chromosomes) Mutations tested No of CF chromosomes Haplotypes Method with the mutation XV2C-KM19 (% of total CF alleles) Exon 3: G85E 4 (033) 3C HinfI/ASO394delTT 2 2B PAGEExon 4: R117H 1 B ASOY122X 2 2C MseI/sequenceI148T 1 B ASO621+IG-J* 1 B MseIIASOExon 5: 711+1G--T 8(07) 8A ASOExon 7: AF311 1 C PAGE/sequencelO78delT 5 (0-42) 5C PAGE/ASOR334W 5 (0-42) 2A,2C,ID MspIlASOR347P 5 (042) 5A CfoI/NcoIR347H 1 Cfol/sequenceExon 9: A455E 1 B ASOExon 10: S492F I C DdeI/sequenceQ493X 1 D ASOl609deICA 1 C PAGE/Ddel/sequenceA1507 3 (025) 3D PAGE/ASOAF508 827 (69) 794B,30D,2C,IA PAGEl677delTA 1 A PAGE/sequenceExon I11: 1717-IG--.A 16(1-3) 14B Modified primers + AvaIIG542X 40 (3-3) 29B,5D,2A Modified primers + BstNiS549R(T--*G) 2 2B ASOG551D 3 (025) 3B HincII/Sau3AR553X 10(0-8) 6A,1B,2C,ID Hincll/sequenceExon 12: 1898+IG--A 1 C ASO1898+ IG-C 2 IC ASOExon 13: l9l8deIGC 1 A PAGE/sequence1949de184 I C PAGE/sequenceG628R(G-+A) 2 2A Sequence2118de14 I c PAGE/sequence2143de1T 1 B PAGE/modified primers2184de1A+2183A--*G 11 (0-9) lIB PAGE/ASO2184de1A 1 ASOK710X 3 (025) IC XmnI2372de18 1 B PAGE/sequenceExon 15: S945L 1 C TaqlExon 17b:L1065P I MnlIL1077P 1 A ASOY1092X 3 (025) 2C,IA Rsal/ASOExon 19: RI1162X 6 (0-5) 5C,IA DdeI/ASO3659delC 3 (025) 3C ASOExon 20: G1244E 2 2A MboIIS1251N 2 2C RsaI3905insT 4 (0-33) 4C PAGE/ASOW1282X 18 (105) 15B,1D MnlI/ASOR1283K 1 C Mnll/sequenceExon 21: N1303K 22 (1-8) 18B,lA,ID Modified primers+BstNI 47 mutations 1031 (85 9) least one CF chromosome (table): 21 of them are very rare as they were found on only one CF chromosome in our population.
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ABCC7 p.Pro574His 7525963:21:357
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PMID: 7521710 [PubMed] Ravnik-Glavac M et al: "Sensitivity of single-strand conformation polymorphism and heteroduplex method for mutation detection in the cystic fibrosis gene."
No. Sentence Comment
121 1078delT (35), L327R (Ravnik-Glavac a al., unpublished), R334W (36), D36K (31), R347L (26), R347P (14), A349V (26), R352Q (30), 1221delCT (34); Exon 8: W401X (31), 1342-1G-C (25); Exon 9: G458V (37), 1525 -1G-A (38); Exon 10: S492F (34), Q493X (39), 1609delCA (40,17), deltaI507 (39,41), deltaF5O8 (3), 1717-1G-A (39,42); Exon 11: G542X (39), S549N, G551D, R553X (43), R553Q (44), A559T (43), R560K (Fine et al., pers. comm.), R560T (39); Exon 12: Y563N (39), 1833delT (Schwartz et al., pers. comm.), P574H (39), 1898 + 1G-C (31), 1898+3A-G (Ferrari et al., pers. comm.); Exon 13: G628R(G-C) (31), Q685X (Firec et al., pers. comm.), K716X (26), L719X (Dork etal., pers. comm.), 2522insC (15), 2556insAT (45), E827X (34); Exon 14a: E831X (Ffrec et al., pers. comm.), R851X (29), 2721delll (31), C866Y (Audrezet et al., pers. comm.); Exon 14b: 2789+5G-A (Highsmith et al., pers. comm.); Exon 15: 2907denT (21), 2991del32 (Dark and TQmmler, pers. comm.), G970R (31); Exon 16: S977P, 3100insA (D6rk et al., pers. comm.); Exon 17a: I1005R (Dork and TQmmler, pers. comm.), 3272-1G-A (46); Exon 17b: H1054D (F6rec et al., pers. comm.), G1061R (Fdrec et al., pers. comm.), 332Oins5, R1066H, A1067T (34), R1066L (Fe"rec etal., pers. comm.), R1070Q (46), E1104X (Zielenski el al., pers. comm.), 3359delCT (46), L1077P (Bozon « a/., pers. comm.), H1085R (46), Y1092X (Bozon etal., pers. comm.), W1098R, M1101K (Zielenski et al., pers. comm.); Exon 18: D1152H (Highsmith et al., pers. comm.); Exon 19:R1162X (36), 3659delC (39), 3662delA (25), 3667del4 (Chillon et al., pers. comm.), 3737ddA (35), 3821ddT (15), I1234V (35), S1235R (31), Q1238X (26), 3849G-A (25), 385O-3T-G (38); Exon20:3860ins31 (Chillon etal., pers. comm.), S1255X (47), 3898insC (26), 3905insT (Malik et al., pers. comm.), D127ON (48), W1282X (49), Q1291R (Dork et al., pers. comm.), Exon 21: N1303H (35), N13O3K (50), W1316X (43); Exon 22: 11328L/4116delA (Dork and TQmmler, pers. comm.), E1371X (25); Exon 23: 4374+ 1G-T (38); Exon 24: 4382delA (Claustres et al., pers. comm.).
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ABCC7 p.Pro574His 7521710:121:501
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PMID: 7682984 [PubMed] Guillermit H et al: "A novel mutation in exon 3 of the CFTR gene."
No. Sentence Comment
63 Other missense mutations reported by Kristidis et al. (1991) as being located in the transmembrane domain, i.e. R117H (exon 4), R334W (exon 7), R347P (exon 7), A455E (exon 9) and P574H (exon 12), are associated with pancreatic sufficiency; these observations are consistent with the genetic hypothesis that pancreatic sufficiency is a 235 dominant phenotypic trait associated with about 10% of the CF alleles.
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ABCC7 p.Pro574His 7682984:63:179
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PMID: 7678316 [PubMed] Kubesch P et al: "Genetic determinants of airways' colonisation with Pseudomonas aeruginosa in cystic fibrosis."
No. Sentence Comment
71 The NBF gene mutations in the study population were all severe disease alleles with respect to pancreatic function, and none of the rare PS alleles G551S, Y563N, P574H was detected.4,25 Hence, our findings do not necessarily imply that a NBF mutation should a priori be considered a "high risk" allele but rather that the more common "severe" disease alleles cluster in the NBF.
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ABCC7 p.Pro574His 7678316:71:162
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PMID: 1279852 [PubMed] Tsui LC et al: "The spectrum of cystic fibrosis mutations."
No. Sentence Comment
122 This hypothesis has been well supported by analysis of patients, which shows that individuals with one or two copies of the missense alleles such as Rll7H, R334W, R347P, A455E (Ref. 12) or P574H (Ref. 12) are found to be PS, whereas those with two copies of nonsense, frameshift, splice-site or a subset of the missense mutations are invariably PI TIGNOVEMBER1992 VOt.
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ABCC7 p.Pro574His 1279852:122:189
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PMID: 1376016 [PubMed] Kristidis P et al: "Genetic determination of exocrine pancreatic function in cystic fibrosis."
No. Sentence Comment
10 This result is thus consistent with the hypothesis that PI and PS in CF are predisposed by the genotype at the CFTR locus; the PS phenotype occurs in patients who have one or two mild CFTR mutations, such as R117H, R334W, R347P, A455E, and P574H, whereas the PI phenotype occurs in patients with two severe alleles, such as AF508, A1507, Q493X, G542X, R553X, W1282X, 621 + 1G-PT, 1717-1G--'A, 556delA, 3659delC, I148T, G480C, V520F, G551D, and R560T.
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ABCC7 p.Pro574His 1376016:10:240
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58 Intron 10: 1717-1G-'A Exon 11: G542X .......... S549R ........... G551D .......... R553X .......... R560T .......... Exon 12: Y563N .......... P574H .......... Exon 19: 3659delC ....... Exon 20: W1282X ....... Exon 21: N1303K ..... G460-C A deletion G482-'A A deletion T575-C 621 + 1G-T C1132-T C1172- G C1496-A G1505-'T G1570-T C1609-T 3-bp deletion 3-bp deletion G1690-T G1717-1-A G1756-T T1779-G G1784- A C1789-T G1811-C T1819- A C1853- A C deletion G3978-A C4041-G Asp 110-His Frameshift Arg 117-His Frameshift Ile 148-Thr Splice mutation Arg 334-Trp Arg 347-Pro Ala 455- Glu Gly 458-'Val Gly480-Cys Gln 493- stop del of Ile 507 del of Phe 508 Val 520-Phe Splice mutation Gly 542- stop Ser 549-'Arg Gly 551-WAsp Arg 553- stop Arg 560- Thr Tyr 563- Asn Pro 574-His Frameshift Trp 1282-stop Asn 1303-Lys Dean et al. 1990 White et al. 1991 Dean et al. 1990 Zielenski et al. 1991a F. Rininsland, D. Bozon, and L.-C. Tsui, unpublished data Zielenski et al. 1991a Gasparini et al. 1991 Dean et al. 1990 Kerem et al. 1990b Cuppens et al. 1990 Strong et al. 1991 Kerem et al. 1990b Kerem et al. 1990b Kerem et al. 1989b Jones et al. 1991 Kerem et al. 1990b Kerem et al. 1990b Kerem et al. 1990b Cutting et al. 1990 Cutting et al. 1990 Kerem et al. 1990b Kerem et al. 1990b Kerem et al. 1990b Kerem et al. 1990b Vidaud et al. 1990 Osborne et al. 1990 PI or PS, but not with both.
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ABCC7 p.Pro574His 1376016:58:143
status: NEW
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ABCC7 p.Pro574His 1376016:58:756
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66 As shown in table 3, meconium ileus Table 2 1181 Table 3 Frequency of 25 CF Mutations in Chromosomes of the Toronto Study Population Mutation AF508 ...... G551D...... G542X...... 621 +1G-'T N1303K..... W1282X..... R1 17H...... 1717-1G-~A R560T...... A1507 ...... R553X...... V52OF ...... R334W ..... A455E...... I148T ...... Q493X...... P574H...... R347P ...... SS6delA ..... 3659delC .... G480C...... 444delA ..... D110H...... G458V...... S549R ...... Y563N......
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ABCC7 p.Pro574His 1376016:66:339
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73 Complete CF Genotypes for 394 Patients No. OF PATIENTS GENOTYPE WITH Allele 1 Allele 2 pla P AF508 ...... AF508 277 (49) 2 G551D 21 (1) 0 G542X 18 (9)c 0 621+1G-~T 11 (1) 0 AI507 7 (1) 0 N1303K 6 (1) 0 R560T 5 0 1717-lG-A 5 (1) 0 556delA 3 0 Q493X 3 0 R553X 3 (1) 0 W1282X 3 0 3659delC 2 0 1148T 1 0 R117H 0 9 A445E 0 2 P574H 0 2 R347P 0 1 G551D ..... 1717-lG-~A 2 0 621+1G-~T 1 0 G480C 1 0 G551D 1 0 V520F 1 (1) 0 G542X ..... V520F 1 0 1148T ...... W1282X 1 (1) 0 W1282X .... W1282X 1 0 N1303K .... R553X 1 (1) 0 R117H ..... R117H 0 1 G542X 0 1 R334W ..... R334W 0 1 a1 Numbers in parentheses are number of patients with neonatal meconium ileus.
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ABCC7 p.Pro574His 1376016:73:320
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81 Table 4 Classification of CF Gene Mutations as Severe or Mild with Respect to Pancreatic Function Type of Mutation Severe (location) Mild (location) Missense (point mutation) ...... 1148T (exon 4) R117H (exon 4) G480C (exon 9) R334W (exon 7) VS2OF (exon 10) GSS1D (exon 11) R347P (exon 7) RS60T (exon 11) A455E (exon 9) N1303K (exon 21) P574H (exon 12) Single amino acid deletion ........ AFS08 (exon 10) A1507 (exon 10) Stop codon (nonsense) ..... Q493X (exon 10) G542X (exon 11) R553X (exon 11) W1282X (exon 20) Splice junction ... 621 + 1G-T (intron 4) 1717-1G-T (intron 10) Frameshift ........ 556delA (exon 4) 3659delC (exon 19) with any of the mild mutations was associated with PS.
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ABCC7 p.Pro574His 1376016:81:337
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85 Accordingly, the mutations R117H, R334W, R347P, A455E, and P574H may be regarded as mild, whereas AF508, AI507, Q493X, G542X, R553X, W1282X, 621 + 1G-T, 1717-1G--A, 556delA, 3659delC, 1148T, G480C, V520F, GSS1D, and R560T are severe.
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ABCC7 p.Pro574His 1376016:85:59
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PMID: 1372093 [PubMed] Cuppens H et al: "Simultaneous screening for 11 mutations in the cystic fibrosis transmembrane conductance regulator gene by multiplex amplification and reverse dot-blot."
No. Sentence Comment
19 Frequency of 31 mutations in the CFTR gene in 194 Belgian CF chromosomes The 51255X, W1316X ;5 S549N, G551D, R553X, A559T;6 D110H, R117H, R347P;' Q493X, S5491, S549R(T-+G), R560T, Y563N, P574H ;9 W846X, Y913C;10 2556insAT;" R334W;" S549R(A-+C);'6 444delA, 3821deIT;" 621 +1G-*T18 mutations were not present in this random sample of the Belgian CF population .
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ABCC7 p.Pro574His 1372093:19:187
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PMID: 15463806 [PubMed] Vankeerberghen A et al: "The cystic fibrosis transmembrane conductance regulator: an intriguing protein with pleiotropic functions."
No. Sentence Comment
390 Mutations that cause a small defect in maturation but normal or increased chloride transport activity, like A455E and P574H tend to be classified in this fifth class of mutations w162x.
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ABCC7 p.Pro574His 15463806:390:118
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PMID: 16963320 [PubMed] Perez MM et al: "CFTR gene analysis in Latin American CF patients: heterogeneous origin and distribution of mutations across the continent."
No. Sentence Comment
78 At least another 38 mutations have been searched for, but none of them were found in the CF patients from Latin America: p.E60X, p.Y122X, p.G178R, p.G330X, p.R347H, p.R352Q, p.S364P, p.A455E, p.Q493X, p.V520F, p.C524X, p.R560T, p.Y563D, p.P574H, p.K710X, p.Q890X, p. R1158X, p.S1196X, p.S1255X, p.D1270N, p.W1310X, p. W1316X, c.405+1G-A, c.444delA, c.556delA, c.574delA, c.1677delTA, c.2043delG, c.2307insA, c.2909delT, c.3120G-A, c.3358delAC, c.3662delA, c.3750delAG, c.3791delC, c.3821delT, c.3849+4A-G, c.3905insT.
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ABCC7 p.Pro574His 16963320:78:239
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PMID: 20166764 [PubMed] Becq F et al: "Cystic fibrosis transmembrane conductance regulator modulators for personalized drug treatment of cystic fibrosis: progress to date."
No. Sentence Comment
105 [38] Class V mutations (e.g. A455E, P574H) are responsible for the production of functional proteins with normal Cl-channel activity and regulation, but at a reduced rate of synthesis.
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ABCC7 p.Pro574His 20166764:105:36
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PMID: 24727426 [PubMed] Wang Y et al: "Understanding how cystic fibrosis mutations disrupt CFTR function: from single molecules to animal models."
No. Sentence Comment
2031 Even more notable was the CF-PS mutation P574H, which disrupted CFTR processing, but formed a "super" normal channel, distinguished by greatly prolonged, but less frequent channel openings compared to wild-type CFTR (Sheppard et al., 1995; Champigny et al., 1995).
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ABCC7 p.Pro574His 24727426:2031:41
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PMID: 24816489 [PubMed] Favia M et al: "Trimethylangelicin promotes the functional rescue of mutant F508del CFTR protein in cystic fibrosis airway cells."
No. Sentence Comment
46 Fischer rat thyroid (FRT) epithelial cells, stably coexpressing human F508del CFTR and the high-sensitivity halide-sensing green fluorescent analog yellow fluorescent protein (HS-YFP) YFP-H148Q/ I152L (FRT-F508del) or stably transfected with a plasmid coding for P574H-CFTR and carrying the resistance gene for zeocin (FRT-P574H) were a generous gift from Dr. L. J. Galietta (Gaslini Institute, Genoa, Italy).
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ABCC7 p.Pro574His 24816489:46:263
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ABCC7 p.Pro574His 24816489:46:323
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47 FRT-F508del and FRT-P574H cells were grown in Coon`s modified Ham`s F-12 medium plus 10% FBS, L-glutamine, and penicillin/streptomycin at 37&#b0;C under 5% CO2.
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ABCC7 p.Pro574His 24816489:47:20
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279 Last, to analyze if the correction effect of TMA was specific for the F508del mutation of CFTR, TMA corrector activity was tested on FRT cells stably expressing P574H-CFTR (FRT-P574H), a mutant of CFTR that, like F508del CFTR, is retained at the ER and can be rescued by incubation for 24 h at reduced temperature (42).
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ABCC7 p.Pro574His 24816489:279:161
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ABCC7 p.Pro574His 24816489:279:177
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280 FRT-P574H cell monolayers were incubated for 24 h with 100 nM TMA or at 27&#b0;C, and the chloride efflux in response to FSK plus IBMX was measured as described in MATERIALS AND METHODS.
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ABCC7 p.Pro574His 24816489:280:4
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287 produce a significant correction in FRT-P574H cells [0.0005 afe; 0.0003 èc;(F/F0)/min, n afd; 4], with respect to the positive low-temperature rescue response [0.011 afe; 0.001 èc;(F/F0)/min, n afd; 5], suggesting that the TMA correction effect is specific for F508del mutation of CFTR.
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ABCC7 p.Pro574His 24816489:287:40
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321 Importantly, we found that TMA corrector activity is specific for the F508del mutation since TMA incubation fails to activate the temperature-sensitive mutant P574H-CFTR (42).
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ABCC7 p.Pro574His 24816489:321:159
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333 J. Zabner for providing CuFi-1 cells, Dr. L. J. Galietta for providing FRT cells stably coexpressing human F508del CFTR and high-sensitivity halide-sensing GFP analog YFP H148Q/ I152L and the P574H-CFTR FRT cells, and Dr. E. Ficker for providing the mutated constructs of hERG.
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ABCC7 p.Pro574His 24816489:333:192
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