ABCMdb

ABCC7 p.Tyr89Cys

ClinVar:	c.266A>G , p.Tyr89Cys ? , not provided
CF databases:	c.266A>G , p.Tyr89Cys (CFTR1) ? , This mutation was detected by DGGE analysis followed by direct sequencing in a young Italian male (19-yr.). His other CF mutation is unknown. It was found once out of 800 CF chromosomes. It was also absent in 200 chromosomes of patients affected by diffuse bronchiectasies or by nasal polyposis and in 100 control chromosomes. (Italian origin)Clinical features of the patient: Bronchiectasies of left lower lobe leading to lobectomia at 8 yrs. Nasal polyposis. Pancreatic sufficiency. Moderate-severe lung disease (FVC 68%, FEVI 69% of predicted) with chronic Pseudomonas lung infection. Oligospermia and teratospermia. Sweat test is normal (<30 mEq/L)(repeated).
Predicted by SNAP2:	A: D (63%), C: N (53%), D: D (80%), E: D (80%), F: N (78%), G: D (75%), H: D (66%), I: N (61%), K: D (85%), L: N (66%), M: N (53%), N: D (75%), P: D (91%), Q: D (75%), R: D (80%), S: D (71%), T: D (71%), V: N (57%), W: N (78%),
Predicted by PROVEAN:	A: D, C: D, D: D, E: D, F: N, G: D, H: D, I: N, K: D, L: N, M: N, N: D, P: D, Q: D, R: D, S: D, T: D, V: N, W: N,

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[hide] Gender-sensitive association of CFTR gene mutation... Mol Hum Reprod. 2005 Aug;11(8):607-14. Epub 2005 Aug 26. Morea A, Cameran M, Rebuffi AG, Marzenta D, Marangon O, Picci L, Zacchello F, Scarpa M
Gender-sensitive association of CFTR gene mutations and 5T allele emerging from a large survey on infertility.
Mol Hum Reprod. 2005 Aug;11(8):607-14. Epub 2005 Aug 26., [PMID:16126774]

Abstract [show]
Human infertility in relation to mutations affecting the cystic fibrosis transmembrane regulator (CFTR) gene has been investigated by different authors. The role of additional variants, such as the possible forms of the thymidine allele (5T, 7T and 9T) of the acceptor splice site of intron 8, has in some instances been considered. However, a large-scale analysis of the CFTR gene and number of thymidine residues, alone and in combination, in the two sexes had not yet been addressed. This was the aim of this study. Two groups were compared, a control group of 20,532 subjects being screened for perspective reproduction, and the patient group represented by 1854 idiopathically infertile cases. Analyses involved PCR-based CFTR mutations assessment, reverse dot-blot IVS8-T polymorphism analyses, denaturing gradient gel electrophoresis (DGGE) and DNA sequencing. The expected 5T increase in infertile men was predominantly owing to the 5/9 genotypic class. The intrinsic rate of 5T fluctuated only slightly among groups, but some gender-related differences arose when comparing their association. Infertile men showed a significantly enriched 5T + CFTR mutation co-presence, distributed in the 5/9 and 5/7 classes. In contrast, females, from both the control and the infertile groups, showed a trend towards a pronounced reduction of such association. The statistical significance of the difference between expected and observed double occurrence of 5T + CFTR traits in women suggests, in line with other reports in the literature, a possible survival-hampering effect. Moreover, regardless of the 5T status, CFTR mutations appear not to be involved in female infertility. These results underline the importance of (i) assessing large sample populations and (ii) considering separately the two genders, whose genotypically opposite correlations with these phenomena may otherwise tend to mask each other.

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76 This test involved nine subjects from the infertile group, revealing the occurrence of the following rare mutations: E217G, T1054A, W356X, D443Y and 3667insTC in males and L997F and R297Q in females and 29 subjects from the control, in which we found: A1009T, D110Y, E826K, G1069R, G1130A, G194V, I556V, L320F, M348K, M82V, P1290T, R117C, R352W, R74W, S42F, S660T, S911R, S912L, T1086A, T582S, V920L and Y89C. Login to comment

[hide] N-terminal CFTR missense variants severely affect ... Hum Mutat. 2008 May;29(5):738-49. Gene GG, Llobet A, Larriba S, de Semir D, Martinez I, Escalada A, Solsona C, Casals T, Aran JM
N-terminal CFTR missense variants severely affect the behavior of the CFTR chloride channel.
Hum Mutat. 2008 May;29(5):738-49., [PMID:18306312]

Abstract [show]
Over 1,500 cystic fibrosis transmembrane conductance regulator (CFTR) gene sequence variations have been identified in patients with cystic fibrosis (CF) and related disorders involving an impaired function of the CFTR chloride channel. However, detailed structure-function analyses have only been established for a few of them. This study aimed evaluating the impact of eight N-terminus CFTR natural missense changes on channel behavior. By site-directed mutagenesis, we generated four CFTR variants in the N-terminal cytoplasmic tail (p.P5L, p.S50P, p.E60K, and p.R75Q) and four in the first transmembrane segment of membrane-spanning domain 1 (p.G85E/V, p.Y89C, and p.E92K). Immunoblot analysis revealed that p.S50P, p.E60K, p.G85E/V, and p.E92K produced only core-glycosylated proteins. Immunofluorescence and whole cell patch-clamp confirmed intracellular retention, thus reflecting a defect of CFTR folding and/or trafficking. In contrast, both p.R75Q and p.Y89C had a glycosylation pattern and a subcellular distribution comparable to the wild-type CFTR, while the percentage of mature p.P5L was considerably reduced, suggesting a major biogenesis flaw on this channel. Nevertheless, whole-cell chloride currents were recorded for all three variants. Single-channel patch-clamp analyses revealed that the channel activity of p.R75Q appeared similar to that of the wild-type CFTR, while both p.P5L and p.Y89C channels displayed abnormal gating. Overall, our results predict a major impact of the CFTR missense variants analyzed, except p.R75Q, on the CF phenotype and highlight the importance of the CFTR N-terminus on channel physiology.

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116 Moreover, some nascent/immature CFTR protein present in the ER and A B 501 E92K Y89C G85E/V P5L S50P E60K R75Q NBD1 NBD2 R FIGURE 1. Login to comment
156 Confocal images from representative xy sections taken from1of 3 independent experiments show the subcellular distribution of wild-type CFTR (WT), p.F508del mutant (F508del), and variants p.S50P (S50P), p.E60K (E60K), p.G85E (G85E), p.G85V (G85V), p.E92K (E92K), p.P5L (P5L), p.R75Q (R75Q), and p.Y89C (Y89C). Login to comment
182 C:Whole-cell currents evoked by variants p. R75Q (R75Q), p.P5L (P5L), and p.Y89C (Y89C).Wild-type CFTR current (WT) is also shown for comparison. Forskolin was applied for 90s. Login to comment
3 By site-directed mutagenesis, we generated four CFTR variants in the N-terminal cytoplasmic tail (p.P5L, p.S50P, p.E60K, and p.R75Q) and four in the first transmembrane segment of membrane-spanning domain 1 (p.G85E/V, p.Y89C, and p.E92K). Login to comment
6 In contrast, both p.R75Q and p.Y89C had a glycosylation pattern and a subcellular distribution comparable to the wild-type CFTR, while the percentage of mature p.P5L was considerably reduced, suggesting a major biogenesis flaw on this channel. Login to comment
8 Single-channel patch-clamp analyses revealed that the channel activity of p.R75Q appeared similar to that of the wild-type CFTR, while both p.P5L and p.Y89C channels displayed abnormal gating. Login to comment
40 The eight CFTR variants included in this study: p.P5L, p.S50P, p.E60 K, p.R75Q, p.G85E, p.G85V, p.Y89C, and p.E92K (Fig. 1A) were generated by oligonucleotide-directed mutagenesis in pCMVCFTRNot6.2wt using the QuickChangeTM XL-Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA) according to the manufacturer`s instructions (see Supplementary Table S1 for a detailed description of the mutagenesis primers employed; available online at http://www.interscience.wiley.com/jpages/1059-7794/supp mat). Login to comment
76 The average capacitances of HEK 293 cells transfected with wild-type CFTR and each of the three active channel variants were: wild-type CFTR (1671 pF; n 5 24); p.R75Q (2473 pF; n 5 18); p.Y89C (2072 pF; n 5 6); and p.P5L (2173 pF; n 5 20). Login to comment
102 RESULTS Description and Cross-Species Analysis of Natural N-Terminus CFTR Variants We chose eight naturally occurring sequence variants, four located across the N-terminal CFTR tail (p.P5L, p.S50P, p.E60K, and p.R75Q), and four within the first segment of MSD1 (p.G85E, p.G85V, p.Y89C, and p.E92 K) (Fig. 1A; Table 1). Login to comment
109 Some of the variants such as p.R75Q and p.Y89C, presented a glycosylation status comparable to the wild-type CFTR, suggesting that they might reach the PM. Login to comment
118 B: Alignment of the N-terminus (amino acids 1 to 100) of the CFTR protein derived from di¡erent species.The sequences derived from human (Homo sapiens, Gen- BankNM_000492), mouse (Mus musculus,GenBankNM_021050), Norway rat (Rattusnovergicus,GenBankNM_031506), European rabbit (Oryctolagus cuniculus, GenBank NM_001082716), cow (Bos taurus, GenBank NM_174018), sheep (Ovis aries, GenBank NM_001009781), African clawed frog (Xenopus laevis, GenBank X65256), and spiny dog'sh (Squalus acanthias, GenBank M83785) were aligned using the ClustalW multiple sequence alignment program.The amino acid residue a¡ected by each of the variants analyzed (p.P5L, p.S50P, p.E60K, p.R75Q, p.G85E, p.G85V, p.Y89C, and p.E92K) is indicated in bold. Login to comment
120 Likewise, variants p.R75Q and p.Y89C (Fig. 3) presented a localization pattern similar to that of wild-type CFTR, with most of the protein visualized in the PM (yellow colocalization of the merge images). Login to comment
131 From the variants expressed in the PM (Fig. 3), only p.R75Q and p.Y89C generated cAMP-stimulated currents comparable to that of wild-type CFTR channel (Fig. 4C). Login to comment
133 Genotype^Phenotype Correlation in the N-Terminal CFTR MissenseVariants Under StudyÃ Missense varianta Phenotype Second allele (number of patients)b p.P5L CF p.F508del (1), p.P205S (1) p.S50P CBAVD p.F508del (1), p.E115del (1) p.E60K CF p.G542X (1), p.I507del (1) p.R75Q HT p.F508del (3), p.E725K (1) B p.R347H (1), p.R75Q (1), n.i. (4) Br c.1584G4A (2), c.1210-7_1210-6delTT (1), n.i.(3) NT p.F508del (1) CP c.1584G4A (1), n.i. (3) MI n.i. (1) CUAVD n.i. (2) OZ n.i. (2) Normal p.R75Q (1), c.2052_2053insA (1), n.i. (1) p.G85E CF p.F508del (8), p.G542X (2), p.I507del (1), c.580-1G4T (1), p.G85E (1), c.1477_ 1478delCA (1) CBAVD p.G576A (1) HT p.L997F (1),WT (1) p.G85V CF p.F508del (2), p.G542X (2), p.Y1092X (1), c.265715G4A (1), p.A1006E, c.1210-7_1210- 6delTT (1), n.i. (1) p.Y89C CF n.i. (1)c p.E92K CF p.F508del (2), p.Q890X (1), p.L206W (1) CBAVD c.1210-7_1210-6delTT (1) ÃThe recommendations for mutation nomenclature (www.hgvs.org/mutnomen/) were used to name CFTR gene sequence variations at both the nucleotide level and the protein level. Login to comment
151 Similar values were observed for p.R75Q (98755 pA/pF, n 5 18), and p.Y89C (114754 pA/pF, n 5 6), indicating that these CFTR variants were trafficked to the PM. Login to comment
158 In the £uorescent images from variants p.P5L, p.R75Q, and p.Y89C, the contour plot from a successfully transfected cell, obtained from the corresponding phase contrast image, is shown to aid the identi'cation of the cell boundary. Login to comment
166 Single-Channel Analysis of N-Terminus CFTR Variants Expressed at the Plasma Membrane To further investigate whether variants p.R75Q, and p.Y89C affected the chloride channel function of CFTR and, particularly, to delineate whether the reduced current density of p.P5L could be attributed only to a folding/trafficking defect or also to a channel malfunction, single-channel experiments were carried out. Login to comment
197 As explained above, variants p.R75Q and p.Y89C were also able to reach the PM and supported chloride currents (Fig. 4). Login to comment
199 Finally, the p.Y89C CFTR was at some extent, FIGURE 5. Login to comment
213 Four of the variants (p.P5L, p.S50P, p.E60 K, and p.R75Q) are localized within the cytosolic N-terminal tail, and the remaining four (p.G85E, p.G85V, p.Y89C, and p.E92K) are embedded in three positions within the first transmembrane segment (TM1) of MSD1. Login to comment
218 Conversely, we have observed that three CFTR missense variants, two (p.P5L and p.R75Q) located in the cytosolic N-terminus and one (p.Y89C) located within the first segment of MSD1, appeared to mature and reach the PM, albeit with different efficiencies. Login to comment
228 Regarding the p.Y89C variant, based on the structural features of the amino acids involved (the bulky hydrophobic tyrosine residue is substituted by the smaller hydrophilic cysteine residue) and its position within TM1 (between the severe folding mutations p.G85E/V and p.E92K) it would also be expected a major structural rearrangement and folding defect of MSD1 conferred by this variant. Login to comment
229 Surprisingly, p.Y89C CFTR was able to mature quite efficiently and to reach the PM. Login to comment
232 Nevertheless, additional work is necessary to accurately define the electrophysiological properties of the p.Y89C variant. Login to comment
255 Genotype^Phenotype Correlations All selected N-terminus CFTR missense variants except p.Y89C, described in an Italian patient [Padoan et al., 2000], have been identified in Spanish individuals with CF or different CFTR-related disorders (see Table 1 and references therein). Login to comment

[hide] Atypical cystic fibrosis and CFTR-related diseases... Clin Rev Allergy Immunol. 2008 Dec;35(3):116-23. Paranjape SM, Zeitlin PL
Atypical cystic fibrosis and CFTR-related diseases.
Clin Rev Allergy Immunol. 2008 Dec;35(3):116-23., [PMID:18493878]

Abstract [show]
Cystic fibrosis (CF), which is among the most common life-shortening recessive illnesses, is caused by mutations of the CF transmembrane conductance regulator (CFTR) and typically involves chronic infection and progressive obstruction of the respiratory tract as well as pancreatic exocrine insufficiency. Disease severity, to some extent, correlates with organ sensitivity to CFTR dysfunction and to the amount of functional protein, which is influenced by the type of mutation. Atypical CF represents approximately 2% of affected individuals, and includes cases presenting in adolescence or adulthood with pancreatic exocrine sufficiency, normal or borderline sweat chloride concentrations, or with a single predominant clinical feature. This review briefly describes diagnostic methods and phenotypic characteristics of classic and atypical CF, as well as CFTR-related diseases, conditions in which mutated CFTR may contribute to the pathogenesis but do not strictly fit established diagnostic criteria.

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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. Login to comment