ABCMdb

ABCC7 p.Arg1070Trp

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II-III (maturation defect, gating defect) <a href="https://www.ncbi.nlm.nih.gov/pubmed/26823392">Veit et al.</a>
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Publications

PMID: 10875853 [PubMed] Casals T et al: "Heterogeneity for mutations in the CFTR gene and clinical correlations in patients with congenital absence of the vas deferens."

No. Sentence Comment

96 L206W/- 5T/9T 1 L206W/3121-1G→A 7T/9T 1 L206W/1949del84 7T/9T 1 transrectal ultrasonography was significantly smaller in∆E115/S50P 7T/7T 1 2869insG/R1070W 7T/7T 1 CBAVD than in CUAVD (F ϭ 8.1, P ϭ 0.005). Login to comment

PMID: 11101688 [PubMed] Jezequel P et al: "Molecular screening of the CFTR gene in men with anomalies of the vas deferens: identification of three novel mutations."

No. Sentence Comment

47 a sweat chloride level of Ͼ70 mmol/l did not show any pulmonary Except for R1070W in exon 17b (four out of 47 ϭ 8.5%)or gastrointestinal symptoms of CF. Login to comment
50 total, ∆F508, T5 allele, R117H, R1070W and L375F representNone had undergone abdominal ultrasonography or excretory 83% of the mutation types in our cohort. Login to comment
54 The three men with CUAVD wereTotal genomic DNA was isolated from the patients` peripheral blood cells and analysed for mutations in the whole CFTR region and splice compound heterozygotes (G542X/R1070W, ∆F508/R117H, junctions. Login to comment
60 Tworegions (1, 2, 5, 6a, 6b, 7, 10, 11, 12, 14a, 14b, 15, 16, 17a, 17b, 18, 20, 22, 23, 24) were amplified with a GC-clamp primer and six exons mutations were found in 31.9% of patients, 31.9% had one Table I. Summary of the clinical and biological findings of a population of men with congenital bilateral absence of the vas deferens (CBAVD, n ϭ 37), congenital unilateral absence of the vas deferens (CUAVD, n ϭ 3) and obstructive azoospermia (Obs A, n ϭ 7) Patient Phenotype Surgical Age Weight Height Sweat test Other clinical CFTR exploration (years) (kg) (m) (Cl- mEq/l) manifestation genotype 1 CBAVD ϩ 40 63 1.72 72 ∆F508/T5 2 CBAVD ϩ 31 66 1.76 40 L1227S/3272-26A→G 3 CBAVD ϩ 29 ∆F508/T5 4 CBAVD 29 sinusitis -/- 5 CBAVD 32 50 1.60 ∆F508/T5 6 CBAVD 35 64 1.66 ∆F508/T5 7 CBAVD ϩ 28 ∆F508/R117H 8 CBAVD ϩ 34 69 1.80 24 ∆F508/R117H 9 CBAVD ϩ 35 65 1.70 R117H/T5 10 CBAVD ϩ 32 50 1.70 31 asthma ∆F508/T5 11 CBAVD ϩ 26 left hydrocele T5/- 12 CBAVD ϩ 23 left varicocele, G551D/T5 asthma, anosmia 13 CBAVD ϩ 29 ∆F508/T5 14 CBAVD ϩ 36 63 1.64 52 ∆F508/R117H 15 CBAVD ϩ 37 60 1.76 ∆F508/T5 16 CBAVD ϩ 34 70 1.65 24 ∆F508/A1067V 17 CBAVD 35 61 1.73 42 ∆F508/R117H 18 CBAVD 25 72 1.82 86 2183AA→G/T5 19 CBAVD 28 88 1.76 7 -/- 20 CBAVD ϩ 29 ∆F508/T5 21 CBAVD 31 48 epididymite -/- 22 CBAVD 28 ∆F508/T5 23 CBAVD ϩ 32 68 1.76 36 flatulence ∆F508/R1070W 24 CBAVD ϩ 31 64 1.76 39 R1162X/T5 25 CBAVD 30 17 asthma R117H/L375F 26 CBAVD ϩ 36 62 1.70 ∆F508/R1070W 27 CBAVD 30 6 -/- 28 CBAVD 35 85 1.70 R1070W/- 29 CBAVD 39 bronchectasis -/- 30 CBAVD ϩ 29 ∆F508/- 31 CBAVD 31 bronchectasis, -/- deafness 32 CBAVD ϩ 26 asthma, otitis -/- 33 CBAVD ϩ 28 allergy -/- 34 CBAVD 37 36 R117H/- 35 CBAVD 33 -/- 36 CBAVD ϩ 30 64 1.68 R117H/T5 37 CBAVD ϩ 37 71 1.78 31 pancreatitis, 621ϩ1G→T/I980K alcoholism 38 CUAVD 43 62 1.68 40 allergy G542X/R1070W 39 CUAVD ϩ 35 allergy ∆F508/R117H 40 CUAVD ϩ 34 hydrocele L375F/G551D 41 Obs A ϩ 32 26 T5/- 42 Obs A 23 60 sinusitis ∆F508/2789ϩ2insA 43 Obs A ϩ 25 80 sinusitis, chronic ∆F508/4428insGA 44 Obs A ϩ 30 bronchitis -/- anosmia 45 Obs A 29 50 -/- 46 Obs A 29 75 1.77 ∆F508/T5 47 Obs A ϩ 30 82 1.66 -/- mutation and the T5 allele, 10.7% had only one mutation and clinical palpation. Login to comment
72 In addition, he had an elevated sweat test (80 mEq/l)39 ∆F508/R117H (TG)10T9/(TG)11T7 23 ∆F508/R1070W (TG)10T9/(TG)10T7 and sinusitis. Login to comment
73 This mutation creates a stop codon 43 nucleotides 26 ∆F508/R1070W (TG)10T9/(TG)10T7 downstream leading to the deletion of 33 C-terminus amino 16 ∆F508/A1067V (TG)10T9/(TG)10T7 acids of the CFTR protein including the TRL-COOH domain.42 ∆F508/2789ϩ2insA (TG)10T9/(TG)10T7 43 ∆F508/4428insGA (TG)10T9/(TG)11T7 This highly conserved proteic site is a perfect match for the 25 R117H/L375F (TG)10T7/(TG)10T7 binding consensus domain of the Naϩ-Hϩ exchanger regulatory 38 G542X/R1070W (TG)10T9/(TG)11T7 factor (NHE-RF), a cytoplasmic phosphoprotein that may play40 L375F/G551D (TG)10T7/(TG)10T7 37 621ϩ1G→T/I980K (TG)10T9/(TG)10T9 an important regulatory role in CFTR function (Wang et al., 2 L1227S/3272-26A→G (TG)10T9/(TG)12T7 1998). Login to comment
79 Under these circumstances, polymorphisms could 28 R1070W/- (TG)11T7/(TG)10T7 be ruled out. Login to comment

PMID: 12167682 [PubMed] Groman JD et al: "Variant cystic fibrosis phenotypes in the absence of CFTR mutations."

No. Sentence Comment

71 MUTATION IDENTIFIED BY SCREENING FOR COMMON MUTATIONS MUTATION IDENTIFIED BY DNA SEQUENCING NO. OF PATIENTS ∆F508 5T* 3 ∆F508 D1152H 2 ∆F508 2789+2insA 2 ∆F508 R117C 2 ∆F508 D110H 1 ∆F508 2789+5G→A 1 ∆F508 P205S 1 ∆F508 L967S 1 ∆F508 I1027T 1 ∆F508 L206W 1 ∆F508 T1053I and 5T 1 ∆F508 V920M and 5T 1 ∆F508 R1070W 1 ∆F508 D579G 1 ∆F508 P67L 1 ∆F508 2811G→T†‡ 1 G85E F191V† 1 R117H G103X and 5T 1 I148T I556V 1 G542X R1162L 1 W1282X D1152H 1 None L138ins and 3272-26 A→G 1 None G463D† and 5T 1 None F693L and 5T 1 ∆F508 None 6 G551D None 1 W1282X None 1 None 5T 4 None 2307insA 1 None L997F 1 None V520I 1 None None 30 in Subject II-2 in Family 1. Login to comment

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. Login to comment

PMID: 12955726 [PubMed] Feldmann D et al: "CFTR genotypes in patients with normal or borderline sweat chloride levels."

No. Sentence Comment

8 R117H, D1152H, L206W, 3272-26A>G, S1235R, G149R, R1070W, S945L, and the poly-T tract variation commonly called IVS8-5T were also observed. Login to comment
44 Table 1 : Genotypes and Phenotypes of Patients with Normal or BordIerline Sweat Tests Patient Age at diagnosis (years) CFTR GENOTYPE* Allele 1 Allele 2 SWEAT CL- MEAN (MMOL/L) PHENOTYPE 1 0.2 F508del G149R 38 P+PI, neonatal hypertrypsinemia, 2 0.3 G551D R117H-7T 31 neonatal hypertrypsinemia 3 0.4 F508del R1070W 30.5 neonatal hypertrypsinemia 4 0.4 F508del R117H-7T 52 P 5 0.6 F508del 3849+10kbC>T 48 P 6 0.11 F508del S945L 58 P+PI 7 1 F508del 5T 40 P+CBAVD 8 2 F508del L206W 53 P 9 2 W1282X 5T 42.5 P 10 5 F508del 3849+10kbC>T 55.5 P 11 5 F508del L206W 55 P 12 5 G91R 5T 47.5 P 13 6 G551D S1235R+5T 49.5 P, neonatal hypertrypsinemia 14 7 F508del 3849+10kb 50 P, nasal popyposis 15 13 F508del R117H-7T 58 P, nasal polyposis 16 18 F508del 5T 60.5 P 17 20 G542X 3849+10kbC>T 52 P+PI 18 21 I507del 3849+10kbC>T 54 P, bronchiectasis 19 30 R347P 3849+10kbC>T 43 P, Pseudomonas colonisation 20 30 I507del L206W 57.5 CBAVD, chronic cough 21 31 F508del R117H-7T 60 CBAVD 22 32 G542X 3849+10kbC>T 30 P, Pseudomonas colonisation 23 34 F508del 3272-26A>G 64 P, CBAVD 24 37 R1070Q D1152H 56 CBAVD, bronchectasis 25 46 F508del D1152H 43 P 26 55 F508del D1152H 48 P, Pseudomonas colonisation 27 56 I507del S1235R 53 P 28 >18 F508del D1152H 60 P+PI 29 >20 F508del 3849+10kbC>T 18 P, bronchiectasis 30 >20 F508del 3272-26A>G 61 P *All mutations are named in accordance with the numbering used in the CFTR Mutation Database: http://www.genet.sickkids.on.ca/cftr/. Login to comment
101 The other mutations observed in trans of severe mutations were G149R, R1070W, S945L and S1235R. Login to comment
102 G149R and R1070W mutations have been previously described in CBAVD patients [Mercier et al., 1995; Jezequel et al., 2000] and S1235R have been described associated with variable pulmonary symptoms and occasionally borderline sweat tests [Monagham et al., 2000]. Login to comment

PMID: 15070876 [PubMed] Dayangac D et al: "Mutations of the CFTR gene in Turkish patients with congenital bilateral absence of the vas deferens."

No. Sentence Comment

44 1 (1.0) DoÈrk et al. 1997b M952I Exon 15 G®C at 2988 Amino acid substitution 1 (1.0) Girodon et al. 1995 3120+1G®A Intron 16 G®A at 3120+1 Aberrant splicing 1 (1.0) Macek et al. 1997 3272-26A®G Intron 17a A®G at 3272±26 Aberrant splicing 1 (1.0) Fanen et al. 1992 R1070W Exon 17b C®T at 3340 Amino acid substitution 1 (1.0) Macek et al. 1993* G1130A Exon 18 G®C at 3521 Amino acid substitution 1 (1.0) This study Mutations were designated following the recommended nomenclature (Beaudet and Tsui, 1993) except for the IVS8-(T)n alleles which are named according to the number of their residual thymidines (Chu et al., 1993). Login to comment
47 *The following mutations were previously reported as personal communications to the CF Genetic Analysis Consortium (http://www.genet.sickkids.on.ca): 359insT by Claustres M, Desgeorges M, Romey M-C; R334Q by FeÂrec C, Quere I, Verlingue C, Raguenes O, AudreÂzet M-P, Mercier B; T388M by Zielenski J, Markiewicz D, Tsui L-C, Rawashdeh M, Khateeb M; E831X by FeÂrec C, Quere I, Audrezet MP, Verlingue C, Guillermit H, Mercier B; M952I by Girodon E, Costes B, Cazeneuve C, Ghanem N, Goossens M; R1070W by Macek M Jr, Sedriks S, Kiesewetter S, Cutting GR; D1152H by Highsmith WE Jr, Burch L, Friedman KJ, Wood BM, Spock A, Silverman LM, Knowles MR. Login to comment
72 CFTR genotypes in 51 patients with congenital bilateral absence of the vas deferens Mutation genotypes IVS8-(TG)mTn M470V n (%) Two mutations detected: D1152H/D1152H (TG)11 7T/ (TG)11 7T V/V 5 (9.8) IVS8-5T/IVS8-5T (TG)13 5T/ (TG)13 5T M/M 2 (3.9) (TG)12 5T/ (TG)13 5T M/V 1 (1.9) (TG)12 5T/ (TG)12 5T V/V 1 (1.9) IVS8-5T/D1152H (TG)12 5T/ (TG)11 7T V/V 2 (3.9) IVS8-5T/DF508 (TG)12 5T/ (TG)10 9T M/V 2 (3.9) IVS8-5T/2789+5G®A (TG)12 5T/ (TG)10 7T M/V 2 (3.9) IVS8-5T/365insT (TG)13 5T/ (TG)11 7T M/V 1 (1.9) IVS8-5T/D110H (TG)12 5T/ (TG)11 7T M/V 1 (1.9) IVS8-5T/E585X (TG)12 5T/ (TG)10 7T M/V 1 (1.9) IVS8-5T/2752-15C®G (TG)12 5T/ (TG)11 7T V/V 1 (1.9) IVS8-5T/M952I (TG)12 5T/ (TG)10 7T M/V 1 (1.9) IVS8-5T/3120+1G®A (TG)12 5T/ (TG)11 7T V/V 1 (1.9) D1152H/A349V (TG)10 7T/ (TG)11 7T M/V 1 (1.9) D1152H/2789+5G®A (TG)10 7T/ (TG)11 7T M/V 1 (1.9) D1152H/G1130A (TG)10 7T/ (TG)11 7T M/V 1 (1.9) CFTRdele2(ins186)/ IVS8-6T (TG)13 6T/ (TG)11 7T M/V 1 (1.9) CFTRdele2(ins186)/D110H (TG)11 7T/ (TG)11 7T V/V 1 (1.9) E831X/D110H (TG)11 7T/ (TG)11 7T V/V 1 (1.9) E831X/1677delTA (TG)11 7T/ (TG)11 7T V/V 1 (1.9) R334Q/R347H (TG)11 7T/ (TG)11 7T V/V 1 (1.9) 1767del6/1767del6 (TG)11 7T/ (TG)11 7T V/V 1 (1.9) 3041-15T®G/3041-15T®G (TG)12 7T/ (TG)12 7T M/M 1 (1.9) 3041-13del7/3041-13del7 (TG)10 7T/ (TG)10 7T M/M 1 (1.9) R1070W/3272-26A®G (TG)10 7T/ (TG)11 7T M/V 1 (1.9) I853F/L997F (TG)11 7T/ (TG)10 9T V/V 1 (1.9) One mutation detected: L997F/? Login to comment

PMID: 15880796 [PubMed] Kerem E et al: "Pharmacological induction of CFTR function in patients with cystic fibrosis: mutation-specific therapy."

No. Sentence Comment

58 C-D565G II DF508 D1507 S549R S549I S549N S549R S945D S945L H1054D G1061R L1065P R1066C R1066M L1077P H1085R N1303K G85E III G551D S492F V520F R553G R560T R560S Y569D IV R117H, R117C, R117P, R117L D1152H, L88S, G91R, E92K, Q98R, P205S, L206W, L227R, F311L, G314E, R334W, R334Q, I336K, T338I, L346P, R347C, R347H, R347L, R347P, L927P, R1070W, R1070Q V 3849 þ 10 kb C ! Login to comment

PMID: 16189704 [PubMed] McGinniss MJ et al: "Extensive sequencing of the CFTR gene: lessons learned from the first 157 patient samples."

No. Sentence Comment

72 The two deletions Table 2 Atypical CF or nonclassic patients in whom extensive sequencing revealed two CFTR mutations Patient Genotype Phenotype Sex Age (years) Sweat chloride concentration (mmol/l) 1 p.S912L/DF508 Chronic lung and sinus disease F 52 Not done 2 p.R1070W/p.N1303K Recurrent respiratory infections F 4.5 2X intermediate 3 p.G551D/c.2789+2 InsA Pancreatic insufficiency, little lung involvement F 50 92, 96 4 c.3849+10kb C>T/p.L732X Failure to thrive, chronic cough, chronic sinusitis M 5.5 70,73 5 p.W1282X/p.R170H Chronic pancreatitis, CBVAD M 44 Borderline (c.1641 AG>T, and c.2949-2953 del TACTC) are expected to be severe disease-associated mutations, since they change the CFTR reading frame; the two patients harboring these novel deletions had a diagnosis of CF with elevated sweat chloride levels and carried a second, previously described, CF mutation. Login to comment

PMID: 18597042 [PubMed] Mornon JP et al: "Atomic model of human cystic fibrosis transmembrane conductance regulator: membrane-spanning domains and coupling interfaces."

No. Sentence Comment

259 Among these, several missense mutations have been observed for the critical R1070 residue (R1070W, R1070Q, R1070P; http:// www.genet.sickkids.on.ca/cftr/), but no functional data are available for them. Login to comment

PMID: 18951463 [PubMed] Krasnov KV et al: "Localization studies of rare missense mutations in cystic fibrosis transmembrane conductance regulator (CFTR) facilitate interpretation of genotype-phenotype relationships."

No. Sentence Comment

1 Mutations of the arginine residue at codon 1070 have been associated with different disease consequences; R1070P and R1070Q with ''severe`` pancreatic insufficient cystic fibrosis (CF) and R1070W with ''mild`` pancreatic sufficient CF or congenital bilateral absence of the vas deferens. Login to comment
4 Confocal microscopy and biotinylation studies revealed that R1070P was not inserted into the apical membrane, R1070W was inserted at levels reduced from wild-type while R1070Q was present in the apical membrane at levels comparable to wild-type. Login to comment
5 The abnormal localization of CFTR bearing R1070P and R1070W was consistent with deleterious consequences in patients; however, the profile of CFTR R1070Q was inconsistent with a ''severe`` phenotype. Login to comment
27 Patients with R1070W (c.3208C4T; p.Arg1070Trp) have pancreatic sufficient CF or congenital bilateral absence of the vas deference (CBAVD) and a normal life span, whereas patients with R1070Q (c.3209G4A; p.Arg1070Gln) and R1070P (c3209G4C; p.Arg1070Pro) have classic CF with significant clinical features of lung disease, pancreatic insufficiency, and elevated sweat chloride levels [Mickle et al., 2000]. Login to comment
28 Previous studies revealed that the biogenesis and function of CFTR bearing R1070W and R1070P is substantially altered, consistent with their pathologic role. Login to comment
29 However, CFTR with R1070Q had a less severe channel gating abnormality than R1070W and either no defect [Seibert et al., 1996; Mickle et al., 2000] or a milder defect than R1070W upon protein processing [Seibert et al., 1996]. Login to comment
30 These functional studies indicate that the R1070Q mutation should cause a milder phenotype than R1070W, rather than the more severe phenotype observed. Login to comment
36 While CFTR bearing R1070W and R1070P displayed localization defects consistent with their associated phenotypes, R1070Q was difficult to distinguish from wild-type CFTR. Login to comment
85 RESULTS Wild-Type CFTR Is Localized toApical Membranes of MDCK Cells To determine the effect of R1070 mutations on CFTR localization, we expressed heterologous CFTR (wild-type or 1 of 3 CFTR mutants-R1070P, R1070Q, or R1070W) from an FRT integration site in MDCK type II cell lines (Fig. 1A). Login to comment
104 CFTR bearing R1070W, a mutation associated with mild disease, localized to the apical surface of MDCK cells (see overlap with apical surface marker in Fig. 2; R1070W, left column) but was also present in other locations in the cell. Login to comment
105 The absence of colocalization with Na1 /K1 ATPase basolateral marker is consistent with presence of CFTR R1070W in the cytoplasm. Login to comment
117 CFTR with R1070W was present in both mature (C band) and immature (B band) forms (Fig. 3A; lane 3), although the ratio of C band to B band and the ratio of C band to actin control (Fig. 3A; lane 3, lower panel) was lower than seen for wild-type CFTR. Login to comment
118 Biotinylation revealed that CFTR R1070W is inserted into the apical membrane, albeit at very low levels when compared to wild-type (Fig. 3B; lane 5). Login to comment
119 These results indicate that R1070W is able to process into mature apically-localized protein but with decreased efficiency compared to wild-type. Login to comment
120 As shown by confocal microscopy, these biochemical assays are consistent with only a fraction of R1070W at the apical membrane. Login to comment
121 There was a small amount of B band in the apical biotinylated fraction of R1070W, implying that some immature CFTR protein bearing this mutation may be inserted into the apical membrane. Login to comment
127 Each panel shows MDCK-FRT cells expressing either wild-type, R1070P, R1070W, or R1070Q CFTR. Login to comment
133 Altered Function of CFTR Bearing R1070W and R1070P Is ConsistentWith Phenotype of Patients CarryingThese Mutations A worldwide survey identified 29 patients who carried the R1070W mutation (24 of whom had detailed clinical information); 26 patients with R1070Q (16 of whom had detailed data), and 2 patients with R1070P (1 of whom had detailed data). Login to comment
136 Out of 24 patients who carry the R1070W mutation, 16 had F508del (c.1521_1523delCTT; p.Phe508del) on the other allele and all were diagnosed with either pancreatic-sufficient CF or CBAVD (Table 1). Login to comment
137 Pancreatic status was unavailable for 6 out of the 16 patients with the R1070W and F508del genotype. Login to comment
140 As for the remaining R1070W patients (8/24 with detailed clinical information), five individuals carried a CFTR mutation associated with pancreatic insufficiency (2869insG (c.2737insG, p.Tyr913fs); G542X (c.1624G4 T, p.Gly542X); R668C-K710X (c. Login to comment
144 The one remaining individual with the R1070W/R668C-K710X genotype had pancreatic-insufficient CF. Login to comment
145 Overall, the R1070W mutation was associated with either pancreatic-sufficient CF or CBAVD when in trans with a CFTR mutation that confers pancreatic insufficiency, indicating that the R1070W mutation generally conferred mild disease. Login to comment
152 A: Western blotting of MDCK cell lysates expressing wild-type, R1070Q, R1070W, or R1070P CFTR. Login to comment
156 CFTR R1070W has a lower amount of stable protein expressed and is readily visible as both C and B bands (lane 3). Login to comment
159 B:Western blotting of the apical biotinylated fraction (odd-numbered lanes) or total lysates (even-numbered lanes) of MDCK cells expressing wild-type, R1070Q, R1070W, or R1070P CFTR.Wild-type CFTR (lanes 1 and 2) and CFTR R1070Q (lanes 3 and 4) have a 205-kDa band in the biotinylated fraction that is consistent with the presence of mature protein in the apical membrane.The fraction of R1070Q inserted into the apical membrane is similar to wild-type.CFTR R1070W (lanes 5 and 6) has a faint 205-kDa band and a very faint 175-kDa band, suggesting that mature and some immature protein is inserted in the apical membrane.CFTR R1070P (lanes 7 and 8) has no bands visible in the biotinylated fraction and only a 175-kDa band is seen in the total lysate, indicating that this mutant protein is not present in the apical membrane.The lower panel is an immunoblot of each apical fraction and each total lysate after incubation with an actin antibody. Login to comment
162 Summarized Clinical Information on R1070 Patients Patient mutations R1070W R1070P R1070Q R1070Q in cis S466X Number of patientsa 24 2 5 11 Second mutaiton dF508 16 1 0 7 other 8 1 5 4 Disease diagnosis CBACD (infertility) 15 0 3 0 Nonclassic CF 9 1 1 0 Classic CF 1 1 1 11 Pancreatic status Su/cient 9 0 1 0 Insu/cient 4a 1 1 10 Not reported 11b 1 3b 1 Sweat chloride levels Normal or low 12 0 1 0 Elevated460 mmol/L 4 1 1 10 Not reported 8b 1 2b 1 a One patient has classic CF; the other three have normal sweat chloride levels and high FVC values. Login to comment
187 Thus, functional studies were informative for all three R1070 mutations, whereby R1070P and R1070W revealed processing defects that are consistent with their role in disease, while the properties of CFTR bearing R1070Q provoked a reevaluation of the established genotype-phenotype relationship that led to a more plausible explanation for the pathology observed in patients with a glutamine substitution at codon 1070. Login to comment
194 Although CFTR R1070W is primarily associated with male infertility (CBAVD), previously published reports and the current study showed that the tryptophan mutation consistently has decreased protein expression levels and incomplete apical localization. Login to comment
195 CFTR R1070W transiently expressed in nonpolarized HEK-293 cells [Mickle et al., 2000], COS-1 cells [Seibert et al., 1996], and the stable expression in isogenic MDCK cell lines shown here all demonstrated lower levels than wild-type CFTR. Login to comment
197 Partial mislocalization of CFTR R1070W may be due to alteration in the rate of membrane recycling, as has previously been observed with N287Y, a CFTR cytoplasmic loop 2 mutant [Silvis et al., 2003]. Login to comment

PMID: 19707853 [PubMed] Mornon JP et al: "Molecular models of the open and closed states of the whole human CFTR protein."

No. Sentence Comment

33 Of note, recent work has also shown that the R1070P and R1070W mutations lead, respectively, to the absence or to reduced levels of insertion of the protein into the apical membrane of polarized epithelial cells [23], suggesting that these mutations may, as DF508, affect the inter-domain interactions of CFTR. Login to comment

PMID: 20100616 [PubMed] Havasi V et al: "Association of cystic fibrosis genetic modifiers with congenital bilateral absence of the vas deferens."

No. Sentence Comment

68 Portuguese CFTR alleles Spanish CFTR alleles Turkish CFTR alleles 5T 22 F508del 11 5T 20 F508del 14 5T 9 D1152H 14 R334W 5 D443Ya 3 D110H 3 R117H 3 G576Aa 3 F508del 2 S1235R 3 R668Ca 3 3041-11del7 2 N1303K 2 G542X 2 1767del6 2 P205S 2 R117H 2 2789þ5G>A 2 D614G 2 V232D 2 CFTRdele2(ins186) 2 G542X 1 L997F 1 3120þ1G>A 1 L206W 1 H609R 1 G1130A 1 V562I 1 N1303H 1 M952I 1 I507del 1 L206W 1 365insT 1 3272-26A>G 1 3272-26A/G 1 E585X 1 2789þ5G>A 1 L15P 1 2752-15C>G 1 G576Aa 1 R347H 1 R334Q 1 R668Ca 1 2689insG 1 R347H 1 CFTRdele2,3 1 R1070W 1 E831X 1 L1227S 1 I 1027T 1 R1070W 1 E831X 1 3272-26A>G 1 L997F 1 I853F 1 A349V 1 6T 1 Note: CFTR ¼ cystic fibrosis transmembrane conductance regulator. Login to comment

PMID: 20233947 [PubMed] He L et al: "Restoration of domain folding and interdomain assembly by second-site suppressors of the DeltaF508 mutation in CFTR."

No. Sentence Comment

95 Indeed, some substitutions on the CL4 side of this interface such as R1070W have been reported to improve ⌬F508-CFTR maturation (23). Login to comment

PMID: 20522854 [PubMed] Sermet-Gaudelus I et al: "Measurement of nasal potential difference in young children with an equivocal sweat test following newborn screening for cystic fibrosis."

No. Sentence Comment

130 Table 3 Genotypes of the children with HIRT according to the diagnostic score cut-off in the 21 patients with reliable NPD tests; results after extensive genetic analysis CFTR genotypes Diagnosis score >0.27 (8 patients) £0.27 (13 patients) A/A 0 F508del/621+3A/G F508del/Q1291R A/AB F508del/R347H F508del/R117H;T7 W846X/R117C n¼2 F508del/R1070W 2183AA/G/L206W F508del/3272-26A/G F508del/R117H;T7; n¼4 A/D 0 F508del/R933G G551D/R352Q B/D G622D/3849+45G/A 0 A/0 F508del/0 n¼2 0 0/0 3 0 0, no identified mutation; A, CF-causing mutation; B, mutation associated with cystic CFTR-related disorders; C, mutation with no clinical consequence ; D, mutation of unknown or uncertain clinical relevance; AB, mutation that is associated with a wide phenotypic spectrum that might belong to either group A or B. CFTR, cystic fibrosis transmembrane conductance regulator; HIRT, hypertrypsinaemia; NPD, nasal potential difference. Login to comment

PMID: 20538955 [PubMed] Sermet-Gaudelus I et al: "Clinical phenotype and genotype of children with borderline sweat test and abnormal nasal epithelial chloride transport."

No. Sentence Comment

162 CLINICAL CHARACTERISTICS OF CHILDREN WITH EQUIVOCAL DIAGNOSES AND NASAL POTENTIAL DIFFERENCE DIAGNOSTIC SCORE <0.27 Pt Mutation Age (yr) NPD Score Sweat Cl2 Chronic CF Pulmonary Disease CF Pathogens Airway Obstruction CF Lung Imaging FEV1 (%) BMI Others 1 F508del/S977F A-D 8 0.181 43 RLRTI, chronic productive cough S. aureus No Bronchiectasis 80 14.5 No Bronchial thickening Atelectasis 2 0/0 4 0.121 43 No S. aureus Yes Air trapping NA 13 Pancreatic extracts 0-0 Bronchial thickening 3 0/0 15 20.032 46 RLRTI S. aureus, P. aeruginosa Yes Air trapping 74 14 Polyposis 0-0 Bronchiectasis 4 F508del/0 2 20.249 57 RLRTI P. aeruginosa Yes Air trapping NA 16 No A-0 5 N1303K/(TG12)T5 11.8 20.263 47 RLRTI S. aureus, P. aeruginosa No Bronchial thickening ND 20 No A-B 6 F508del/L206W 5.9 20.278 40 RLRTI S. aureus No Bronchial thickening 115 22 Chronic pancreatitis A-AB 7 R668C/0 15 20.403 40 RLRTI None Yes Bronchiectasis 112 20 No B-0 Air trapping 8 F508del/L997F A-B 1 20.594 38 RLRTI, chronic productive cough P. aeruginosa No Bronchial thickening NA 16 CF hepatopathy 9 G576A;R668C/S1235R 8 20.659 31 0 None Wheezing Normal 100 20 No B-B 10 G542X/0 5 20.718 49 RLRTI, chronic productive cough S. aureus No Bronchial thickening NA 18 No A-0 11 0/0 7 20.742 37 RLRTI None No Normal 106 18 No 0-0 12 F508del/D110E 16 20.777 50 No S. aureus No No 100 21 No A-AB 13 F508del/R1070W 7 21.006 40 RLRTI S. aureus Wheezing Bronchial thickening 110 14 No A-AB 14 F508del-L467F/0 12 21.897 55 RLRTI, chronic productive cough S. aureus No Bronchiectasis 109 17 Pansinusitis A-0 15 F508del/H1054D 9 22.327 59 RLRTI, chronic productive cough S. aureus No Bronchial thickening 100 20 DIOS A-D Definition of abbreviations: A, B, AB, and D: A 5 CF-causing mutation; B 5 mutation that results in a CFTR-RD (clinical entities associated with CFTR mutations that do not meet the current diagnostic criteria for CF); AB 5 wide-spectrum mutation that may belong to either group A or group B; D 5 mutation of uncertain clinical relevance; BMI 5 body mass index; CF 5 cystic fibrosis; CFTR 5 gene encoding cystic fibrosis transmembrane conductance regulator; DIOS 5 distal intestinal obstructive syndrome; NA 5 not applicable; ND 5 not determined; NPD 5 nasal potential difference; P. aeruginosa 5 Pseudomonas aeruginosa; Pt 5 patient; RLRTI 5 recurrent lower respiratory tract infection; S. aureus 5 Staphylococcus aureus. Login to comment

PMID: 20667826 [PubMed] Thibodeau PH et al: "The cystic fibrosis-causing mutation deltaF508 affects multiple steps in cystic fibrosis transmembrane conductance regulator biogenesis."

No. Sentence Comment

228 The substitution of R1070W had little effect on the maturation of wild type CFTR but measurably promoted trafficking of ⌬F508 CFTR (Fig. 6B). Login to comment
230 To evaluate the potential mechanisms by which the R1070W mutation rescued ⌬F508, this mutation was also introduced into the ⌬F508-3M and F508K backgrounds (Fig. 6, C and D). Login to comment
232 The combination of the -3M mutations with the R1070W mutation increased ⌬F508 Band C production, as compared with ⌬F508-3M and ⌬F508/ R1070W alone. Login to comment
234 In this regard, the R1070W mutation induced the formation of Band C in the F508K mutant predicted to disrupt the interdomain interaction, an effect not seen for low temperature or with the -3M mutations (Fig. 6D). Login to comment
314 C, the R1070W and -3M suppressor mutations were evaluated for their ability to rescue the ⌬F508 mutation either independently or in combination. Login to comment
315 The inclusion of the -3M and R1070W mutations in combination rescued more ⌬F508 CFTRthaneithersuppressorsetalone.Cellswereculturedat37 °CandevaluatedbyWesternblottingusingthe M3A7 antibody. Login to comment
316 D, trafficking of the F508K missense protein was evaluated with the R1070W mutation. Login to comment
317 Trafficking of F508K was partially rescued by the R1070W mutation. Login to comment
325 The R1070W mutant in TMD2 suppresses ⌬F508 by promoting the interactions between NBD1 and ICL4 as required for maturation. Login to comment
326 By relieving the QC-NBD interactions, the -3M and R1070W mutations promote CFTR maturation; upper pathway. Login to comment
339 Although both the Arg-1070 and -3M suppressors rescue ⌬F508, suppression by the R1070W mutation is likely accomplished by independent mechanisms. Login to comment
341 Thus, the R1070W mutation putatively promotes appropriate domain-domain associations by increasing hydrophobic interactions (affinity) at the NBD1-ICL4 domain-domain interaction surface. Login to comment
342 The relatively hydrophobic surface proximal to the Phe508 position could potentially accommodate the hydrophobicity and volume of the R1070W substitution. Login to comment
344 Maturation of the F508K CFTR molecule was potentially facilitated by interactions between the indole side chain from R1070W and the NBD1 surface. Login to comment
346 Intriguingly, the R1070W mutation, which rescues ⌬F508 CFTR, has been identified in patients with mild disease (congenital bilateral absence of the vas deferens, pancreatic sufficient CF). Login to comment
347 Previous studies suggest that in the wild type background the R1070W mutation alters protein expression, localization, and function (44). Login to comment

PMID: 20717170 [PubMed] Rene C et al: "p.Ser1235Arg should no longer be considered as a cystic fibrosis mutation: results from a large collaborative study."

No. Sentence Comment

69 Of these, eight were compound heterozygous for p.Phe508del, and two for mutations associated with CF-related diseases,3,27 (p.Arg117His;T7) and p.Arg1070Trp (http://www.genet.sickkids.on.ca/cftr/). Login to comment
95 of subjects Allele 1 Allele 2 p.Ser1235Arg;p.Arg785X p.Phe508del Severe CF 2 p.Ser1235Arg;p.Arg785X NAa Severe CF 1 p.Ser1235Arg;875+1G4A (c.743+1C4A) 3629delT (c.3497delT) Severe CF 1 p.Ser1235Arg;p.Arg785X p.Gly542X Severe CF 1 p.Ser1235Arg;(TG)13(T)5 p.Gly551Asp Mild CF 1 p.Ser1235Arg;(TG)13(T)5 p.Phe508del CBAVD 6 p.Ser1235Arg;(TG)13(T)5 p.Arg1070Trp CBAVD 1 p.Ser1235Arg;(TG)13(T)5 p.Arg117His; (T)7 CBAVD 1 p.Ser1235Arg p.Phe508del CBAVD 1 p.Ser1235Arg - CBAVD 1 p.Ser1235Arg;(TG)13(T)5 p.Phe508del CUAVD 1 Suspicion CF/mild phenotype: p.Ser1235Arg - Genital symptoms 5 p.Ser1235Arg - Respiratory symptoms 16 p.Ser1235Arg;(TG)13(T)5 p.Phe508del Respiratory symptoms 2 p.Ser1235Arg 406-6T4C (c.274-6T4C) Respiratory symptoms 1 p.Ser1235Arg p.Tyr1092X Respiratory symptoms 1 p.Ser1235Arg p.Glu831X Respiratory symptoms 1 p.Ser1235Arg p.Gln493X Respiratory symptoms 1 p.Ser1235Arg p.Ile507del Respiratory symptoms 1 p.Ser1235Arg - Digestive symptoms 13 p.Ser1235Arg p.Gly542X Digestive symptoms 1 p.Ser1235Arg - Hyperechogenic fetal bowel 5 p.Ser1235Arg p.Arg668Cys; p.Arg576Ala Hyperechogenic fetal bowel 1 p.Ser1235Arg p.Val920Met Hyperechogenic fetal bowel 1 p.Ser1235Arg p.Phe508del Hyperechogenic fetal bowel 1 aNA: not available; we could only test the mother and a healthy sister (the patient was deceased and the father`s DNA was not available). Login to comment
115 Genotype Familial information Allele 1 Allele 2 1 p.Ser1235Arg; (TG)13(T)5 p.Phe508del Brother of CUAVD (no parental project) 2 p.Ser1235Arg; (TG)13(T)5 p.Phe508del Sister of CUAVD 3 p.Ser1235Arg; (TG)13(T)5 p.Arg1070Trp Sister of CBAVD 4 p.Ser1235Arg p.Gly542X Father of CF [p.Phe508del]+ [p.Gly542X] and healthy children [p.Ser1235Arg]+[(TG)11(T)5] 5 p.Ser1235Arg p.Gln493X Father of CF [p.Phe508del]+ [p.Gln493X] 6 p.Ser1235Arg p.Phe508del Uncle of CF (no parental project) 7 p.Ser1235Arg p.Phe508del Mother of CF [p.Phe508del]+ [1717-1G4A (c.1585-1G4A)] 8 p.Ser1235Arg 2347delG (c.2215delG) Mother of CF [p.Phe508del]+ [2347delG (c.2215delG)] 9 p.Ser1235Arg (TG)11(T)5 Mother of non-CF fetus with hyperechogenic fetal bowel [p.Phe508del]+[(TG)11(T)5] 10 p.Ser1235Arg p.Phe508del Sister of CBAVD 11 p.Ser1235Arg p.Phe508del Sister of CBAVD 12 p.Ser1235Arg p.Glu831X Sister of CF-like patient 13 p.Ser1235Arg p.Phe508del First-cousin of CF-like patient 14 p.Ser1235Arg p.Phe508del A 18-month-old child with prenatal diagnostic based on familial history of CF Table 2 Comparison of the allelic frequencies of p.Ser1235Arg and p.Phe508del in the general population, CF and CBAVD patients Populations Significance (P-values) General population, this study CF patients30 (n¼7420) CBAVD patients30 (n¼1626) General population vs CF General population vs CBAVD p.Phe508del 0.82% (n¼1950) 67.2% 21.6% S (Po0.05) S (Po0.05) p.Ser1235Arg 0.76% (n¼4228) 0.11% 0.43% S (Po0.05) NS (P¼0.17) Significance (P-values) p.Phe508del vs p.Ser1235Arg NS (P¼0.91) S (Po0.05) S (Po0.05) Abbreviations: N, number of unrelated tested chromosomes; S, significant difference; NS, no significant difference. Login to comment

PMID: 20880762 [PubMed] de Prada Merino A et al: "[R74W;R1070W;D1270N]: a new complex allele responsible for cystic fibrosis."

No. Sentence Comment

5 Keywords: Complex allele; R74W; D1270N; R1070W; CFTR; Cystic fibrosis 1. Login to comment
59 To our knowledge, R1070W has never been described within a complex allele. Login to comment
60 R1070W is considered a mutation of "mild" pancreatic-sufficient CF or of CFTR-related disease including CBAVD [1,16]; functional studies have revealed abnormal localization of CFTR bearing R1070W [16]. Login to comment

PMID: 20976528 [PubMed] Kalid O et al: "Small molecule correctors of F508del-CFTR discovered by structure-based virtual screening."

No. Sentence Comment

111 In support of this hypothesis, it has been demonstrated that the double mutant F508del/R1070W, predicted by our model to improve hydrophobic packing in this region (Fig. 7), improves the trafficking of F508del-CFTR (Philip J. Thomas, personal communication). Login to comment
134 a MOLCAD surface of the interface site. b Binding site residues (stick representation) and binding site interaction regions generated with SiteMap. Yellow hydrophobic region, blue H-bond donor region, and red H-bond acceptor region Y1073 F1074 R1070W W496 M498 V510 F1068 Fig. 7 In silico generation of the R1070W mutant in the F508del model suggests that a tryptophan in this position may restore interactions lost in the F508del mutant evaluate this large set of compounds, in vitro screening commenced with a high throughput iodide flux corrector assay [22] performed in the laboratory of Professor Luis Galietta (Advanced Biotechnology Center, Genova, Italy). Login to comment

PMID: 21520952 [PubMed] Loo TW et al: "Benzbromarone stabilizes DeltaF508 CFTR at the cell surface."

No. Sentence Comment

14 It was recently reported that the stability of ΔF508 CFTR at the cell surface approached that of wild-type CFTR when NBDÀTMD2 interactions were restored.21 It was shown that introduction of a V510D mutation into NBD1 promoted the maturation and stability of ΔF508 CFTR by forming a a salt bridge with Arg1070 of TMD2.21 Similarly, maturation of ΔF508 CFTR was promoted by a R1070W suppressor mutation in TMD2.5 These suppressor mutation results suggested that direct binding of a compound to the TMDs may promote the maturation and stability of ΔF508 CFTR. Login to comment

PMID: 21182301 [PubMed] Loo TW et al: "The W232R suppressor mutation promotes maturation of a truncation mutant lacking both nucleotide-binding domains and restores interdomain assembly and activity of P-glycoprotein processing mutants."

No. Sentence Comment

331 For example, the R1070W mutation was postulated to promote folding through hydrophobic interactions with NBD1 in ΔF508-CFTR (65). Login to comment

PMID: 17331079 [PubMed] Alonso MJ et al: "Spectrum of mutations in the CFTR gene in cystic fibrosis patients of Spanish ancestry."

No. Sentence Comment

52 Mutation 0.46-0.35 9 c.1078delT #, p.R347P # 8 p.G85V, c.621 + 1G > T #, p.S549R (T > G) #, p.R553X #, c.3849 + 10kbC > T # 7 p.R347H #, c.1812-1G > A, p.R709X 0.30-0.10 6 p.H199Y, p.P205S, 5 p.R117H #, p.G551D #, p.W1089X, p.Y1092X, CFTR50kbdel 4 c.296 + 3insT, c.1717-1G > A #, c.1949del84, c.3849 + 1G > A 3 p.E92K, c.936delTA, c.1717-8G > A, c.1341G > A, p.A561E, c.2603delT, p.G1244E, [p.D1270N; p.R74W] 2 p.Q2X, p.P5L, CFTRdele2,3, p.S50P, p.E60K, c.405 + 1G > A, c.1677delTA, p.L558S, p.G673X, p.R851X, p.Y1014C, p.Q1100P, p.M1101K, p.D1152H, CFTRdele19, p.G1244V, p.Q1281X, p.Y1381X <0,1 1 c.124del23bp, p.Q30X, p.W57X, c.406-1G > A, p.Q98R, p.E115del, c.519delT, p.L159S, c.711 + 3A > T, p.W202X, c.875 + 1G > A, p.E278del, p.W361R, c.1215delG, p.L365P, p.A399D, c.1548delG, p.K536X, p.R560G, c.1782delA, p.L571S, [p.G576A; p.R668C], p.T582R, p.E585X, c.1898 + 1G > A, c.1898 + 3A > G, c.2051delTT, p.E692X, p.R851L, c.2711delT, c.2751 + 3A > G, c.2752-26A > G, p.D924N, p.S945L, c.3121-1G > A, p.V1008D, p.L1065R, [p.R1070W; p.R668C], [p.F1074L; 5T], p.H1085R, p.R1158X, c.3659delC #, c.3667del4, c.3737delA, c.3860ins31, c.3905insT #, c.4005 + 1G > A, p.T1299I, p.E1308X, p.Q1313X, c.4095 + 2T > A, rearrangements study (n = 4) Mutations identified in CF families with mixed European origin: c.182delT, p.L1254X, c.4010del4. Login to comment
67 Seven other complex alleles were observed: [c.296 + 3insT; p.V754M], [p.F508del; p.I1027T], [p.S549R; -102T > A], [p.G576A; p.R668C], [p.R1070W; p.R668C], [p.D1270N; p.R74W] and [p.T1299I; p.I148T]. Login to comment

PMID: 23071149 [PubMed] Okiyoneda T et al: "Fixing cystic fibrosis by correcting CFTR domain assembly."

No. Sentence Comment

75 Stabilization of the NBD1-MSD2 interface by second site suppressor mutations (e.g., R1070W) largely restored the WT-like coupling efficiency between NBD1 stability and F508-CFTR folding (Rabeh et al., 2012). Login to comment
88 Likewise, reversing the NBD1-MSD2 interface instability by second site mutations (e.g., R1070W) only marginally rescued the F508-CFTR phenotype (Fig. 2 B). Login to comment

PMID: 22680785 [PubMed] Liu X et al: "Thermal instability of DeltaF508 cystic fibrosis transmembrane conductance regulator (CFTR) channel function: protection by single suppressor mutations and inhibiting channel activity."

No. Sentence Comment

5 Thermal inactivation of ΔF508 was mitigated by each of five suppressor mutations, I539T, R553M, G550E, R555K, and R1070W, but each exerted unique effects on the severity of, and recovery from, thermal inactivation. Login to comment
18 We identified unique functional signatures for five second-site mutations, four in NBD1 (I539T, G550E, R553M, and R555K) and one in the fourth intracellular loop (ICL4, R1070W), and also investigated the relation of thermal stability to variations in channel gating brought about by intracellular cAMP, CFTR potentiators, and CFTR inhibitors. Login to comment
150 Protection from thermal inactivation by mutations that increased the open probability of ΔF508 CFTR channels led us to examine the thermal behavior of R1070W/ΔF508 CFTR. Login to comment
151 The substitution of this aromatic side chain in ICL4 increased the open probability of the double mutant 7 and was proposed to act, at least in part, by repairing defective coupling between NBD1 and ICL4.6,7 Results depicted in Figure 7A show that pairing ΔF508 with R1070W improved the thermal stability of the double mutant, but did not restore wt-like thermal stability. Login to comment
154 Pairing R1070W and a second NBD1 suppressor, R555K, with ΔF508, however, resulted in thermal stability that was indistinguishable from that of wt CFTR (Figure 7B). Login to comment
155 In contrast, combining ΔF508 with R1070W and I539T resulted in channels that could not sustain the elevated conductance seen immediately after warming to 37 °C but were nevertheless able to sustain a substantial conductance at 37 °C (Figure 7C). Login to comment
168 Protection of ΔF508 CFTR from thermal inactivation by R1070W. Login to comment
169 (A) Following stimulation, an oocyte expressing R1070W/ ΔF508 CFTR (n = 4) was warmed to 37 °C (gray bar and circles) for 10 min twice in succession. After cooling to 22 °C, the oocyte was exposed to 10 μM CF172 (black bar and circles). Login to comment
170 (B) Following stimulation, an oocyte expressing R555K/R1070W/ΔF508 CFTR (n = 4) was warmed to 37 °C for 10 min twice. After cooling to 22 °C, the oocyte was exposed to 10 μM CF172. Login to comment
171 (C) Following stimulation, an oocyte expressing I539T/R1070W/ΔF508 CFTR (n = 4) was warmed to 37 °C for 10 min twice. After cooling to 22 °C, the oocyte was exposed to 10 μM CF172. Login to comment
247 Similarly, G550E, R555K, and R1070W; when combined individually with ΔF508, improved protein maturation at 37 °C to at most 18% of wt,4,6 but nevertheless significantly improved the thermal stability of the double mutant channels. Login to comment
265 The ICL4 mutation, R1070W, increased the thermal stability of ΔF508 CFTR channel function, although it did not fully restore the wt-like behavior at 37 °C. Login to comment
266 Like G550E, R553M, and R555K, this second-site mutation has been associated with increased open probability of the double mutant,7 an effect attributed to a partial improvement in the interaction between NBD1 and ICL4.29,57 Combining the ICL4 mutation with an NBD1 suppressor mutation on the ΔF508 background (R555K/R1070W/ΔF508), however, fully restored wt-like thermal stability at 37 °C, an "additive" effect similar to that reported by Mendoza et al 6 in their study of the effect of these mutations on NBD1 folding and ΔF508 CFTR protein yield. Login to comment
267 The rescue of wt thermal stability of channel function by the R553M mutation, however, indicates that the structural modifications introduced by combining ΔF508 with R1070W are not required for the thermal stabilization of channel gating. Login to comment
268 Mendoza et al.6 also reported that combining R1070W with I539T, which alone increased the cellular yield of NBD1 to about 80% of wt, resulted in a yield of the I539T/R1070W/ ΔF508 protein that was 76% of the wt level. Login to comment

PMID: 22406676 [PubMed] Aleksandrov AA et al: "Allosteric modulation balances thermodynamic stability and restores function of DeltaF508 CFTR."

No. Sentence Comment

151 Thus, both the constant and varying temperature experiments demonstrated a requirement for a balance between thermal stability and channel activity in CFTR, consistent with considerations of such a relationship between stability and catalytic function of proteins in general.28 NBD1 stabilization restores an NBD1-CL4 interface In addition to destabilizing NBD1, deletion of F508 also disrupts interdomain contacts including the interface between the NBD1 surface and the cytoplasmic loop (CL)4 in MSD2 in which the residue normally participates.29 Specific second-site mutations on either side of this interface (e.g., R1070W or V510D) have been shown to promote maturation of ΔF508 CFTR.27,30,31 The current observations that the ΔF508 protein with NBD1 strongly stabilized by the proline and I539T substitutions had channel activity similar to the WT at physiological temperature suggested either that the NBD1-CL4 interface is not important for function or that it is adequately restored by NBD1 stabilization. Login to comment

PMID: 22265408 [PubMed] Rabeh WM et al: "Correction of both NBD1 energetics and domain interface is required to restore DeltaF508 CFTR folding and function."

No. Sentence Comment

132 Remarkably, NBD1, MSD1, and NBD2 were susceptible to comparable misfolding upon disrupting the NBD1-CL4 interface by the R1070W substitution in WT CFTR. Login to comment
136 R1070W or V510D substitutions at the interfaces restored the proximity of the DF508 NBD1 and CL4 as shown by Cys crosslinking. Login to comment
137 The NBD1-CL4 interface stabilization can be accomplished by filling the cavity created by the DF508 with the bulky hydrophobic side chain of R1070W or by salt bridge formation between V510D in NBD1 and R1070 in CL4 (Figure S6B) (He et al., 2010; Loo et al., 2008, 2010; Thibodeau et al., 2010). Login to comment
138 R1070W, similar to V510D, alone modestly increased the DF508 CFTR folding efficiency and cellular and PM expression (Figures 6A-6E), in part confirming previous reports (He et al., 2010; Thibodeau et al., 2010; Loo et al., 2010). Login to comment
140 At the individual domain level, the combination of R1S and R1070W, but not R1S alone, largely restored domain assembly, as indicated by the WT-like trypsin resistance of the MSD1, MSD2, and NBD2 in DF508-CFTR-R1S-R1070W (Figure 5C). Login to comment
141 Similarly, the DF508 CFTR folding and expression were synergistically rescued by combining the DF508-NBD1 stabilizing mutation DRI with R1070W or V510D (Figures 6F-6H and S7B), ruling out nonspecific effects of second-site mutations. Login to comment
142 Direct energetic stabilization of the DF508-NBD1-0S and -3S by the V510D mutation was marginal (Figure S5). Login to comment
144 Accordingly, substantially increased folding efficiency of DF508-CFTR-1218X was only achieved by synergistic stabilization of the NBD1-CL4 interface (R1070W or V510D) and NBD1 (3S, -3R, or -R1S) (Figures 7A- 7C, and S7C). Login to comment
148 Likewise, disrupting the NBD1-CL4 interface by the R1070W mutation compromised both WT CFTR and CFTR-1218X folding (Figures 6B, 6C, 6E, left panel, and 7C), as well as the NBD1, NBD2, and MSD1 conformation, which was partly rescued by stabilizing the NBD1 with 3S, R, or R1S (Figure S6A). Login to comment
153 Red and blue circles depict the combined effect of R1070W or V5010D interface and 3S, R, or R1S stabilizing mutations on the DF508 CFTR expression. Login to comment
158 resistance of NBD1, MSD1, and NBD2 (Figure S6A) as well as restored folding and processing upon NBD1 stabilization by 3S, R, R1S, or DRI in the CFTR-R1070W and CFTR-1218X- R1070W (Figures 6B, 6C, 6E, and 7C). Login to comment
166 (D and E) Correction of the DF508 CFTR gating defect by the combination of the 3S and R1070W mutations. Login to comment
174 WT-like domain assembly and stabilization of DF508 CFTR were achieved by a combination of NBD1 and NBD1-CL4 interface-stabilizing mutations (e.g., 3S and R1070W or V510D). Login to comment
175 M2* and red line depict the R1070W mutation. Login to comment
178 The 3S and R1070W mutations increased the DF508 CFTR Po by 27% and 54%, respectively. Login to comment
197 Although interdomain interactions likely stabilize NBD1 conformationally in WT CFTR, as suggested by the enhanced NBD1 protease susceptibility upon destabilizing the MSD1 or NBD2 by CF-causing point mutation (Cui et al., 2007; Du and Lukacs, 2009; Du et al., 2005; Xiong et al., 1997), or the NBD1-CL4 interface by R1070W (Figure S6A), these stabilizing factors are compromised due to coupled misfolding of MSD1, MSD2, and NBD2 in DF508 CFTR and other mutants (Cui et al., 2007; Du and Lukacs, 2009; Du et al., 2005; Xiong et al., 1997). Login to comment
203 S, R, or DRI mutations increased the WT channel folding efficiency (by 20%-80%) in proportion with NBD1 stabilization, a tendency that is also observed in the background of NBD1-CL4 destabilization in R1070W-CFTR (Figures 4B and 6B). Login to comment
204 Stabilization of the NBD1-CL4 interface by salt bridge formation between V510D and R1070 also improved WT CFTR biogenesis (Figure 6B). Login to comment
133 Remarkably, NBD1, MSD1, and NBD2 were susceptible to comparable misfolding upon disrupting the NBD1-CL4 interface by the R1070W substitution in WT CFTR. Login to comment
139 R1070W, similar to V510D, alone modestly increased the DF508 CFTR folding efficiency and cellular and PM expression (Figures 6A-6E), in part confirming previous reports (He et al., 2010; Thibodeau et al., 2010; Loo et al., 2010). Login to comment
145 Accordingly, substantially increased folding efficiency of DF508-CFTR-1218X was only achieved by synergistic stabilization of the NBD1-CL4 interface (R1070W or V510D) and NBD1 (3S, -3R, or -R1S) (Figures 7A- 7C, and S7C). Login to comment
149 Likewise, disrupting the NBD1-CL4 interface by the R1070W mutation compromised both WT CFTR and CFTR-1218X folding (Figures 6B, 6C, 6E, left panel, and 7C), as well as the NBD1, NBD2, and MSD1 conformation, which was partly rescued by stabilizing the NBD1 with 3S, R, or R1S (Figure S6A). Login to comment
154 Red and blue circles depict the combined effect of R1070W or V5010D interface and 3S, R, or R1S stabilizing mutations on the DF508 CFTR expression. Login to comment
159 resistance of NBD1, MSD1, and NBD2 (Figure S6A) as well as restored folding and processing upon NBD1 stabilization by 3S, R, R1S, or DRI in the CFTR-R1070W and CFTR-1218X- R1070W (Figures 6B, 6C, 6E, and 7C). Login to comment
167 (D and E) Correction of the DF508 CFTR gating defect by the combination of the 3S and R1070W mutations. Login to comment
176 M2* and red line depict the R1070W mutation. Login to comment
179 The 3S and R1070W mutations increased the DF508 CFTR Po by 27% and 54%, respectively. Login to comment
198 Although interdomain interactions likely stabilize NBD1 conformationally in WT CFTR, as suggested by the enhanced NBD1 protease susceptibility upon destabilizing the MSD1 or NBD2 by CF-causing point mutation (Cui et al., 2007; Du and Lukacs, 2009; Du et al., 2005; Xiong et al., 1997), or the NBD1-CL4 interface by R1070W (Figure S6A), these stabilizing factors are compromised due to coupled misfolding of MSD1, MSD2, and NBD2 in DF508 CFTR and other mutants (Cui et al., 2007; Du and Lukacs, 2009; Du et al., 2005; Xiong et al., 1997). Login to comment

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

103 Table 1. Continued Mutations in 257 patients Allele counts of each mutation % of variant alleles (183) % of all alleles tested (514) R1070W 1 0.55 0.19 R1158X 1 0.55 0.19 R1438W 1 0.55 0.19 R334W 2 1.09 0.39 R352W 1 0.55 0.19 R553X 2 1.09 0.39 R668C 2 1.09 0.39 R74W 3 1.64 0.58 R75X 3 1.64 0.58 S1235R 2 1.09 0.39 S492F 2 1.09 0.39 S549N 1 0.55 0.19 S573CS573C 1 0.55 0.19 S945L 1 0.55 0.19 T351S 1 0.55 0.19 T501A 2 1.09 0.39 T604ST604S 1 0.55 0.19 V11I 1 0.55 0.19 V201 mol/L 1 0.55 0.19 V232D 2 1.09 0.39 V754 mol/L 1 0.55 0.19 W1089X 2 1.09 0.39 W1098C 1 0.55 0.19 W1204X 4 2.19 0.78 Y563N 1 0.55 0.19 Y913XY913X 1 0.55 0.19 85 different mutations 183 100.00 35.60 Novel variants are in boldface, mutations on the ACMG/ACOG panel are italicized. Login to comment
186 Table 3. Continued CFTR mutations Alleles Relative mutation frequency (%) (of 317) G567A 1 Ͻ1 S573C 1 Ͻ1 E585X 1 Ͻ1 T604S 1 Ͻ1 F693L 1 Ͻ1 V754 mol/L 1 Ͻ1 2108delA 1 Ͻ1 2184delA 1 Ͻ1 2215insG 1 Ͻ1 2585delT 1 Ͻ1 2752 - 6TϾC 1 Ͻ1 E831X 1 Ͻ1 D836Y 1 Ͻ1 Y913X 1 Ͻ1 S945L 1 Ͻ1 L967S 1 Ͻ1 3171delC 1 Ͻ1 3199del6 1 Ͻ1 3271 ϩ 8AϾG 1 Ͻ1 R1066H 1 Ͻ1 R1070W 1 Ͻ1 Y1092X 1 Ͻ1 W1098C 1 Ͻ1 3500 - 2AϾT 1 Ͻ1 4016insT 1 Ͻ1 4374 ϩ 13AϾG 1 Ͻ1 D1152H 1 Ͻ1 R1158X 1 Ͻ1 R1162X 1 Ͻ1 W1282X 1 Ͻ1 N1303K 1 Ͻ1 Q1313X 1 Ͻ1 P1372L 1 Ͻ1 R1438W 1 Ͻ1 Total 317 100 Table 3. Login to comment

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

108 g D44G, 300delA, W57X, 405+1G>A, D110H, E116K, 541del4, 542del7, L137R, 621+2T>G, I175V, H199R, H199Y, C225X, V232D, Q290X, E292X, G314V, T338I, 1221delCT, W401X, Q452P, I502T, 1716+2T>C, G544S, R560S, A561E, V562I, Y569D, 1898+3A>G, 1898+5G>A, G628R(G>A), 2143delT, G673X, R851X, Q890X, S977F, 3129del4, 3154delG, 3271+1G>A, G1061R, R1066L, R1070W, 3601-17T>C, S1196X, 3732delA, G1249R, 3898insC, 4374+1G>A, del25kb. Login to comment
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. Login to comment

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

31 2 R1066S (C. Fe´rec, I. Quere, C. Verlingue, O. Raguenes, M.-P. Au- drezet, and B. Mercier, personal communication), F1074L (T. Casals, M. D. Ramos, J. Gime´nez, V. Nunes, and X. Estivill, personal communication), K1060T (T. Casals, M. Chillo´n, V. Nunes, J. Gime´nez, M. D. Ramos, and X. Estivill, personal communication), L1065R (T. Casals, M. D. Ramos, J. Gime´nez, V. Nunes, and X. Estivill, personal communication), T1086I (T. Bienvenu, S. Bousquet, C. Herbulot, C. Beldjord, and J. C. Kaplan, personal communication), and R1070W (M. Macek, S. Sedriks, S. Kiesewetter, and G. R. Cutting, personal communication). Login to comment

PMID: 8662892 [PubMed] Seibert FS et al: "Disease-associated mutations in the fourth cytoplasmic loop of cystic fibrosis transmembrane conductance regulator compromise biosynthetic processing and chloride channel activity."

No. Sentence Comment

64 The mature forms of the other six mutants (F1052V, K1060T, A1067T, G1069R, R1070W, R1070Q) were produced in relatively normal amounts (band C), although for A1067T and R1070W CFTR the ratio of the complex-glycosylated to core-glycosylated bands was significantly lower than for wild-type CFTR. Login to comment
82 A1067T, R1070W, and R1070Q CFTR had significantly lower efflux levels and significantly lower protein expression than wild-type CFTR in COS-1 cells. Login to comment
94 As shown in Fig. 5, B and C, in some mutants this reduction in open probability was associated with a significant reduction in mean burst duration (F1052V, G1069R) or an increase in mean interburst duration (R1070W). Login to comment
128 B: छ, WT; E, G1069R; µ, R1070Q; Ç, R1070W; Ⅺ, control. Login to comment
132 Different substitutions at the same residue always produced the same effect, i.e. R1066C, R1066H, and R1066L, as well as M1101K and M1101R all inhibited maturation, whereas R1070W and R1070Q were both normally processed. Login to comment
142 A, examples of wild-type, F1052V, K1060T, A1067T, G1069R, R1070Q, and R1070W CFTR single channel currents recorded from inside-out membrane patches at a membrane potential of -30 mV. Login to comment
71 The mature forms of the other six mutants (F1052V, K1060T, A1067T, G1069R, R1070W, R1070Q) were produced in relatively normal amounts (band C), although for A1067T and R1070W CFTR the ratio of the complex-glycosylated to core-glycosylated bands was significantly lower than for wild-type CFTR. Login to comment
89 A1067T, R1070W, and R1070Q CFTR had significantly lower efflux levels and significantly lower protein expression than wild-type CFTR in COS-1 cells. Login to comment
101 As shown in Fig. 5, B and C, in some mutants this reduction in open probability was associated with a significant reduction in mean burst duration (F1052V, G1069R) or an increase in mean interburst duration (R1070W). Login to comment
134 B: L, WT; E, G1069R; &#b5;, R1070Q; &#c7;, R1070W; M, control. Login to comment
138 Different substitutions at the same residue always produced the same effect, i.e. R1066C, R1066H, and R1066L, as well as M1101K and M1101R all inhibited maturation, whereas R1070W and R1070Q were both normally processed. Login to comment
148 A, examples of wild-type, F1052V, K1060T, A1067T, G1069R, R1070Q, and R1070W CFTR single channel currents recorded from inside-out membrane patches at a membrane potential of 230 mV. Login to comment

PMID: 8530001 [PubMed] Ferec C et al: "Neonatal screening for cystic fibrosis: result of a pilot study using both immunoreactive trypsinogen and cystic fibrosis gene mutation analyses."

No. Sentence Comment

82 {17bi DI507 [ Y569X W846X 2789+5G->A ,' $492F i ] i I G551D 2622+1 G->A Y1092X 1717-1 G->A E827X A1067T G542X 2183 AA->G R1066H R560K 2184 ins A 3320,ins 5 R553G R1070W 1806 del A & 4005+1G->A W1282X ] i "- Exons Fig.2 Distribution of the different mutations (except AF508) of the CFTR gene in Brittany Table 1 Mutations and genotypes in newborns Genotypes of newborns Number Sweat test AF508/AF508 7 + > 90 AF508/1806 del A 1 + > 90 R553G/G551D 1 Borderline (60) AF508/G551D 1 + > 90 AF508/R1070W 1 40 AF508/G542X 1 + > 90 AF508/G149R 1 45 Total 13 Mutations found in heterozygote newborns AF508 31 R560K 1 1078 del T 1 G544S l G542X 1 V317A 1 R347H 1 V322A 1 Total 38 gene. Login to comment

PMID: 7739684 [PubMed] Chillon M et al: "Mutations in the cystic fibrosis gene in patients with congenital absence of the vas deferens."

No. Sentence Comment

74 OF PATIENTS POLYT GENOTYPE† ⌬F508/R668C ⌬F508/D1152H ⌬F508/D1270N ⌬F508/R75L ⌬F508/R117H ⌬F508/L206W ⌬F508/R258G ⌬F508/S1235R ⌬F508/R347H ⌬F508/R347H R117H/G1349D R117H/712-1G→T G149R/R668C R347H/R1066H R553X/R668C R1070W/2869insG ⌬F508/- G542X/- W1282X/- R334W/- K1060T/- R1162X/- N1303K/- A800G/- ⌬F508/- ⌬F508/- ⌬F508/- ⌬E115/- R117H/- R347H/- G542X/- R553X/- 1677delTA/- 2184delA/- 2789ϩ5G→Α/- S1235R/- W1282X/- -/- -/- -/- -/- 2 2 2 1 1 1 1 1 1 1 1 1 1 1 1 1 22 4 3 1 1 1 1 1 7 1 1 1 1 2 1 1 1 1 1 1 1 3 3 1 19 9T/7T 9T/7T 9T/7T 9T/7T 9T/7T 9T/9T 9T/7T 9T/7T 9T/7T 9T/9T 7T/7T 7T/9T 9T/7T 9T/7T 7T/7T 7T/7T 9T/5T 9T/5T 7T/5T 7T/5T 7T/5T 7T/5T 9T/5T 5T/5T 9T/7T 9T/9T 7T/7T 7T/7T 7T/7T 9T/7T 9T/7T 7T/7T 7T/7T 7T/7T 7T/7T 7T/9T 7T/7T 9T/5T 7T/5T 5T/5T 7T/7T -/- 3 7T/9T *Data were obtained from the Spanish population analyzed in this study. Login to comment

PMID: 7532150 [PubMed] Casals T et al: "Extensive analysis of 40 infertile patients with congenital absence of the vas deferens: in 50% of cases only one CFTR allele could be detected."

No. Sentence Comment

88 A second change in exon 17b corresponded to R1070W (M. Macek Jr., personal communication to CFGAC), which has not previously been detected in Spanish CF chromosomes (Chill6n et al. 1994b). Login to comment
103 In 13 cases, the mutations are known to be associated with severe CF (AF508, G542X, Rl162X and 1677delTA), whereas in five cases, the phenotypic effect of the mutation is still unknown (AEll5, K1060T, R334W, R1070W, and 2789 + 5G---)A); in one case (Rll7H), the mutation is known to result in mild CE Of these mutations, R334W seems to cause pancreatic insufficiency with a variable age of onset (X. Estivill, in press), whereas mutation 2789 + 5G--)A (W. E. Jr. High- smith, personal communication to the CFGAC) has frequently been found in adult CF patients and is probably involved in a mild phenotype (T. Casals et al. unpublished). Login to comment

PMID: 7539448 [PubMed] Le Lannou D et al: "Obstructive azoospermia with agenesis of vas deferens or with bronchiectasia (Young's syndrome): a genetic approach."

No. Sentence Comment

43 Six patients were double heterozygotes: AF508/R117H (three cases), AF508/ R1070W (two cases), AF508/A1067V (one case). Login to comment
50 1 z 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 Details of cystic fibrosis (CF) mutations identified congenital agenesis of vas deferens CF mutation AF5O8/ AF508/ AF508/ AF508/ AF508/ AF5O8/A1067V AF5O8/ AF508/R1070W R117H/ AF508/R1070W - AF508/ R117H/ 621 + 1 G->T7 AF508/RU7H R117H/ AF508/R117H AF5O8/ - AF508/R117H G551D/ R117H/ Respiratory disease - - asthma bronchiectasia - - asthma - - - - rhinitis - - - - - asthma - - in all patients with Sweat tests (mmol/ml of chloride) 72a 42 31 64a 24 - 30 39 17 - 6 - 36 11 31 24 - - 7 52 - - Patients with two results >60 mmol/ml were considered positive. Login to comment
71 an incomplete clinical form of CF. However, if AF508 is excluded, the most frequent mutations in the CAVD group (R117H and R1070W) were not observed in a group of 108 CF patients in a separate study, and these mutations seemed to be specific to CAVD. Login to comment

PMID: 7526685 [PubMed] Morral N et al: "Independent origins of cystic fibrosis mutations R334W, R347P, R1162X, and 3849 + 10kbC-->T provide evidence of mutation recurrence in the CFTR gene."

No. Sentence Comment

108 )-.T R347L Audrezet et al. 1993 G--S-C R347P Dean et al. 1990 1789 ......... C--.G R553G C. Ferec, personal communication CI-T R553X Cutting et al. 1990 1790 ......... G---A R553Q Dork et al.1991a 3328 ......... C-OT R1066C Fanen et al. 1992 3329 ......... G-.A R1066H Ferec et al. 1992 GT R1066L Mercier et al. 1993 3340 ......... CT R1070W M. Macek, Jr., unpublished data 3341 ......... G-A R1070Q Mercier et al. 1993 a This change is a polymorphism, not a disease mutation. Login to comment

PMID: 10762539 [PubMed] Mickle JE et al: "Effects of cystic fibrosis and congenital bilateral absence of the vas deferens-associated mutations on cystic fibrosis transmembrane conductance regulator-mediated regulation of separate channels."

No. Sentence Comment

50 Expression Analysis The mutations DF508, R1070W, D1270N, and G1349D were created in the vector pBQ4.7 containing CFTR cDNA (pBQ4.7 is a gift from J. Rommens and L. Login to comment
51 C. Tsui), by single-stranded mutagenesis (Youssoufian et al. 1995), and then were shuttled into pRSV-CFTR, a Rous sarcoma virus (RSV)-driven expression plasmid, by use of Kpn2I and HpaI (for DF508) or NcoI and SalI (for R1070W, D1270N, and G1349D) restriction sites common to both plasmids (Fulmer et al. 1995). Login to comment
68 Mutations at codon 1070 of TMD2 were selected, since two mutations (R1070P and R1070Q) have been associated with CF, whereas a third (R1070W) has been observed in men with CBAVD (table 1). Login to comment
69 The R1070W mutation was first reported by us to the Cystic Fibrosis Genetic Analysis Consortium. Login to comment
74 The latter mutation is predicted to change the amino acid at residue 1070 from arginine to tryptophan and is designated "R1070W." Login to comment
91 Thus, mutations R1070Q and R1070W altered but did not prohibit complex glycosylation. Login to comment
97 OF CASES PHENOTYPE Lung Status Pancreatic Status Sweat Cl2 Fertility Normala Abnormal Not Reported Sufficient Insufficient Not Reported Reported (Mean 5 SEM [mmol/literb ]) Not Reported Subfertilec Not Reported R1070W (7)d 5 0 2 5 0 2e 6 ( ) 50.2 5 13.4 1 6 1 CBAVD R1070P (2)f 0 1 1 0 1 1 1 (Positive) 1 0 2 CF R1070Q (14)g 0 7 7 0 7 7 7 (Positive) 7 2 12 CF D1270N (9)h 4 0 5 4 0 5 3 ( ) 77.5 5 16.7 6 3 6 CBAVD G1349D (3)i 0 0 3 0 0 3 ) 3 1 2 CF a No history of chronic lung disease. Login to comment
172 B, CBAVD(R1070W)- and CF(R1070P and R1070Q)-associated mutants in TMD2 had I-V plots similar to those of wild-type CFTR: outwardly rectified currents (blackened circles) that responded to DIDS (unblackened circles) and glibenclamide (crosses). Login to comment
180 Thus, the combination of I-V relationship and response to inhibitors allowed dissection of whole-cell Cl-currents into two components: outwardly rectified and DIDS sensitive, carried by separate channels such as ORCCs; and linear, DIDS insensitive, and gli- Table 2 Summary of Processing and Whole-Cell Function of CFTR Mutants DOMAIN AND MUTATION PHENOTYPE CFTR STATUS a Processingb Function Band A Band B Band C Cl2 Channel Regulatoryc Not applicable: Wild type Normal 2 2 111 111 1 NBF1:d A455Ee CFe 1 11 2 111 1 DF508 CF 1 11 2 1 2 G551D CF 2 2 111 1 2 TMD2: R1070W CBAVD 2 1 11 111 1 R1070P CF 1 11 2 111 1 R1070Q CF 2 1 11 111 1 NBF2: D1270N CBAVD 2 2 111 111 1 G1349D CF 2 2 111 111 1 a A minus sign (2) denotes absence; a single plus sign (1) denotes "low"; a double plus sign (11) denotes "intermediate"; and a triple plus sign denotes "high." Login to comment
188 .05 mutant (R1070W) and the CF mutants (R1070P and R1070Q) in TMD2 generated outwardly rectified Cl2 currents that were inhibited by DIDS (fig. 3B). Login to comment
230 Electronic-Database Information Accession numbers and URLs for data in this article are as follows: Cystic Fibrosis Genetic Analysis Consortium, http://www .genet.sickkids.on.ca/CFTR (for R1070W) Online Mendelian Inheritance in Man (OMIM), http://www .ncbi.nim.nih.gov/Omim (for CF [MIM 219700], CFTR [MIM 602421], and CBAVD [MIM 277180]) References Anderson MP, Rich DP, Gregory RJ, Smith AE, Welsh MJ (1991) Generation of cAMP-activated chloride currents by expression of CFTR. Login to comment

PMID: 22265409 [PubMed] Mendoza JL et al: "Requirements for efficient correction of DeltaF508 CFTR revealed by analyses of evolved sequences."

No. Sentence Comment

146 The R1070W sterically clashes with the F508 position in the model (Figure 6A, top) a prediction consistent with fact that it is a CF-causing mutation (Krasnov et al., 2008) that inhibits CFTR maturation when F508 is present (open circle, Figure 6A, bottom). Login to comment
148 The ability of the R1070W mutation to counteract the ICL4-NBD1 interface defect caused by the DF508 mutation, allowed assessment of the quantitative effects of suppression of both defects concurrently. Login to comment
149 When NBD1 suppressor mutations were introduced on top of R1070W and DF508, the slope was restored (m = 0.77, R = 0.47) (Figure 6B). Login to comment
152 Similarly, the R1070W interface mutant produces an 7-fold increase in the slope, but only a modest improvement in CFTR maturation. Login to comment
158 Mirroring the maturation results, correction of either the NBD1 defect (I539T, R555K, red bars) or the ICL4-NBD1 defect (R1070W, white bar) alone provided only modest improvements of function. Login to comment
215 0.0 0.5 1.0 1.5 0 25 50 75 100 WT F WT F Relative Yield CFTR (ELISA) Current Density (pA/pF) F I 5 3 9 T R 5 5 5 K R 1 0 7 0 W I 5 3 9 T R 5 5 5 K W T R 1 0 7 0 W 0.0 0.5 0.0 0.5 1.0 1.5 2.0 2.5 Relative Conductance F F-R1070W D C 0.0 0.5 1.0 1.5 2.0 0 1 WT F Relative Yield NBD1 ( -gal.) 0.0 0.5 1.0 1.5 2.0 0 1 WT F Relative Yield NBD1 ( -gal.) Relative Yield CFTR (ELISA) A + Forskolin/IBMX + Inh 172 60 sec 500 pA + Forskolin/IBMX + Inh 172 B Relative Yield CFTR (ELISA) Figure 6. Login to comment
217 The R1070W (open triangle) mutation in ICL4 modestly improves the relative yield of DF508 CFTR maturation and decreases the relative yield for CFTR wild-type (open circle) consistent with the predicted steric clash with the F508 side chain (top) (&#b1;SEM). Login to comment
219 When R1070W is combined with mutations that improve DF508 NBD1 folding yield, I539T, G550E, R553M, R555K, and 3M (open triangles), the correlation between NBD1 folding and CFTR maturation in the wild-type protein is restored (m = 0.77, R = 0.47, black line) (&#b1;SEM). Login to comment
224 A representative trace of the corrected mutant, DF508-I539T-R1070W CFTR (cyan triangles) is more like wild-type (filled circles) than DF508 (filled triangles) (inset). Login to comment
241 On the WT or DF508 CFTR models, F508K, R1070, R1070W residues were modeled using the PyMOL (Schrodinger, 2010) mutagenesis function (Figures 1B, 2B, rightmost panel, 5, 6, and S1). Login to comment

PMID: 22973227 [PubMed] Patrick AE et al: "Development of CFTR Structure."

No. Sentence Comment

85 Notably, the W277 position is equivalent to the R1070 position in ICL4 (Mornon et al., 2008) that when mutated, R1070W, suppresses the ࢞F508 mutation (Thibodeau et al., 2010; Mendoza et al., 2012). Login to comment

PMID: 23055971 [PubMed] Molinski S et al: "Functional Rescue of F508del-CFTR Using Small Molecule Correctors."

No. Sentence Comment

54 Substitution of the arginine at position 1070 with tryptophan (R1070W) in the context of the Wt-CFTR, introduces a bulky group on the face of the coupling helix that interacts with NBD1 and like the substitutions above, this leads to misprocessing. Login to comment
58 Introduction of R1070W or V510D in the F508del-CFTR protein partially corrects folding of the full-length protein, highlighting the idea that even in the absence of F508, assembly of the CFTR can be partially restored through structural changes at key loci in the protein (Thibodeau et al., 2010; Mendoza et al., 2012). Login to comment
64 However, in combination with R1070W, a mutation that reconstitutes a more Wt-like ICL4: NBD1 interface, the NBD1-"stabilizing" mutants mediate full correction and near normal processing. Login to comment

PMID: 23060795 [PubMed] Pedemonte N et al: "Pharmacological Correctors of Mutant CFTR Mistrafficking."

No. Sentence Comment

145 The second type of mutation, namely R1070W, improves the interaction of NBD1 with CL4 by providing an aromatic group that compensates for the lack of F508. Login to comment

PMID: 23104983 [PubMed] He L et al: "Correctors of DeltaF508 CFTR restore global conformational maturation without thermally stabilizing the mutant protein."

No. Sentence Comment

138 The R1070W substitution in CL4 may do so by contributing to interactions among a cluster of aromatic residues at the interface that is weakened by the absence of F508 from the NBD1 surface (11), whereas the V510D mutation was proposed to provide a salt bridge with R1070 (31). Login to comment
139 The V510D mutant on the NBD1 side of the interface is very sensitive to further enhancement of maturation by the compound, whereas that on the CL4 side (R1070W) responds only rather weakly (Fig. 5B). Login to comment
140 This difference may reflect the fact that the V510D substitution stabilizes isolated NBD1 (32) in the absence of the rest of CFTR, as well as influencing the interface, whereas R1070W has only the latter effect. Login to comment
141 One might speculate that the less substantial influence of VX-809 on R1070W/èc;F508 CFTR could possibly reflect similar effects of either the tryptophan residue or the aromatic small molecule to partially fill the void left by the absence of the F508 residue. Login to comment
142 Interestingly, as has already been observed by others (9), the influence of the combined V510D and R1070W substitutions is similar to that of V510D alone, which would not be expected if V510D were forming a salt bridge with R1070 but might be if V510D acted primarily to stabilize the NBD1 domain, as has been observed (32). Login to comment
149 B) èc;F508 with NBD1/CL4 interface substitutions R1070W and/or V510D. Login to comment
157 Interestingly, the patterns of enhancement by the compounds were remarkably similar for each of the three classes of NBD1 stabilizing mutations (èc;F/èc;RI, èc;F4S, and èc;F/4PT) with or without one of the interface substitutions (R1070W or V510D). Login to comment

PMID: 23378596 [PubMed] Hunt JF et al: "Cystic fibrosis transmembrane conductance regulator (ABCC7) structure."

No. Sentence Comment

255 A second-site R1070W mutation in the proximal region of the TMDs produces significant suppression of the defect (Thibodeau et al. 2010), presumably by increasing hydrophobic interactions with the altered conformation of residue V510 in F508del-NBD1. Login to comment

PMID: 23380248 [PubMed] Hanrahan JW et al: "Novel pharmacological strategies to treat cystic fibrosis."

No. Sentence Comment

145 The two-step folding hypothesis also suggests a rational way to test for new pharmacological chaperones that act at each step: pharmacological chaperones that correct NBD1 conformational stability should be additive, with mutations such as R1070W that restore contacts between NBD1 and MSD2 (CL4). Login to comment

PMID: 23457292 [PubMed] Chong PA et al: "Dynamics intrinsic to cystic fibrosis transmembrane conductance regulator function and stability."

No. Sentence Comment

98 Similar results are seen with an R1070W mutation, which might be expected to enhance the hydrophobic contacts at this interface or fill the void introduced by deletion of F508, or with a combined R1070D/V510R mutation, which would reverse the proposed salt bridge (Farinha et al. 2010; Loo et al. 2010). Login to comment

PMID: 23757197 [PubMed] Galietta LJ et al: "Managing the underlying cause of cystic fibrosis: a future role for potentiators and correctors."

No. Sentence Comment

123 Other mutations, such as p.Arg1070Trp, improve the interaction between NBD1 and CL4 [60]. Login to comment

PMID: 23835419 [PubMed] Loo TW et al: "Corrector VX-809 stabilizes the first transmembrane domain of CFTR."

No. Sentence Comment

180 To test if the V232D or H1085R mutants could be rescued by suppressor mutations in other domains, suppressor mutations in NBD1 (I539T), the NBD1-TMD2 interface (V510D), or TMD2 (R1070W) (only V232D) locations were introduced into the mutants. Login to comment
184 The I539T and R1070W mutations only rescued DF508 CFTR. Login to comment
185 We previously reported that the R1070W mutation also reduces the maturation efficiency of wild-type CFTR [32] while V510D does not [33]. Login to comment
237 (C) Extracts of cells expressing processing mutants DF508, V232D, or H1085R with or without the V510D, I539T, or R1070W suppressor mutations were subjected to immunoblot analysis. Login to comment

PMID: 23890012 [PubMed] Farinha CM et al: "Revertants, low temperature, and correctors reveal the mechanism of F508del-CFTR rescue by VX-809 and suggest multiple agents for full correction."

No. Sentence Comment

16 The second F508del-associated defect impairs CFTR interdomain folding, namely, (1) the NBD1-NBD2 dimerization interface (critical for channel activation and accounting for the F508del-CFTR gating defect; Dalemans et al., 1991), which can be rescued by the G550E revertant, and (2) the interaction of NBD1 with the fourth intracellular loop (ICL4) of TMD2 (Serohijos et al., 2008), shown to be reverted by either V510D (Loo et al., 2010) or R1070W, which both fill the pocket left empty by F508del (Thibodeau et al., 2010). Login to comment
23 Herein, we explored the MoA of VX-809 by analyzing its synergistic/additive effect with those of previously characterized genetic revertants, which rescue F508del-CFTR by causing different effects: 4RK affecting traffic (Roxo-Rosa et al., 2006), G550E (Roxo-Rosa et al., 2006) and R555K increasing channel gating by strengthening the NBD1:NBD2 dimer interface, and R1070W (Serohijos et al., 2008) and V510D (Wang et al., 2007a; Loo et al., 2010) by filling the NBD1:ICL4 interface. Login to comment
36 VX-809 Adds to VRT-325 and Corr-4a to Rescue F508del-CFTR but Exhibits Variable Effects on Genetic Revertants In order to characterize the rescue mechanism of VX-809 on F508del-CFTR, we then tested the effect of incubating it together with VRT-325 and Corr-4a on BHK cells stably expressing this mutant alone or in cis with the following genetic revertants: (1) 4RK (where the four AFTs were simultaneously mutated to lysines), (2) G550E, (3) R1070W, (4) V510D, or (5) R555K (Figures 1A-1E). Login to comment
42 Moreover, whereas both VRT-325 and VX-809 add significantly to the effect of R1070W by 17% (p = 0.011) and 28% (p = 1.6 10 5 ), respectively, Corr-4a does not. Login to comment
46 These analyses were performed for three of the revertants with different F508del-CFTR effects (4RK, traffic; G550E, NBD1:NBD2 dimer interface; and R1070W, NBD1:ICL4 interface). Login to comment
57 Effect of Small Molecule Correctors on F508del-CFTR and Genetic Revertants (A-F) BHK cell lines stably expressing CFTR bearing F508del alone (A) or in cis with 4RK (B), G550E (C), R1070W (D), V510D (E), and R555K (F) were incubated for 24 hr with 6.7 mM VRT-325, 10 mM Corr-4a, or 3 mM VX-809 alone or in combination. Login to comment
72 We then assessed the effect of revertants G550E and R1070W by modeling (Figure 3F; Figures S2B and S2C), whereby G550E allows a salt bridge to form across the ATP binding site with Lys1250 and other residues from NBD2 (Figure 3F). Login to comment
73 By contrast, R1070W (Figure S2A) plausibly fills in the space left empty by absence of F508 at the NBD1:ICL4 interface as previously proposed (Thibodeau et al., 2010). Login to comment
87 Rescue of F508del-CFTR by Low Temperature Is Additive to Genetic Revertants To learn more about how low temperature rescues F508del-CFTR, we assessed its combined effect with that of the above genetic revertants: G550E, R1070W, 4RK, V510D, and R555K (Figures 4A and 4B). Login to comment
88 Results show that low temperature further increases processing levels of F508del-CFTR by the five genetic revertants, namely, V510D, G550E, R1070W, 4RK, and R555K, by an additional 35%, 65%, 38%, 27%, and 22%, respectively (compare gray and black bars in Figure 4B). Login to comment
101 Interestingly, these additive effects were observed not only for revertants promoting protein-autonomous folding (G550E, V510D, and R1070W) but also for the 4RK revertant, which bypasses the AFT-mediated ER retention. Login to comment
102 Combination of Different Genetic Revertants Is Also Additive Next, to assess the full potential for F508del-CFTR rescue, we combined the effects of folding and traffic revertants by producing stable BHK cell lines expressing F508del-G550E-CFTR, where 4RK, V510D, or R1070W were also added in cis, and analyzed processing (Figures 4C and 4D). Login to comment
103 Results in Figure 4C show that 4RK, V510D, and R1070W further increased processing of G550E-F508del-CFTR by another 12%, 59%, and 70%, respectively. Login to comment
104 In fact, the combined effects of G550E with either V510D or R1070W bring F508del-CFTR processing to z80%, i.e., close to levels of WT-CFTR, which can be further increased at 26 C reaching 88%. Login to comment
165 In contrast, the additive effect of VX-809 to R1070W and V510D is rather modest, thus suggesting that this corrector acts more similarly to R1070W/V510D than to G550E/R555K. Login to comment
166 VRT-325 in turn (and comparatively to its modest effect alone) significantly increases (by z29% versus VRT-325 alone) the rescue efficiency of R1070W and also V510D, suggesting effects at distinct sites. Login to comment
193 Furthermore, modeling also predicts that revertants G550E and R1070W act at different CFTR interdomain contacts disrupted by F508del: whereas G550E seems to restore the strength of the NBD1:NBD2 interface, R1070W rather promotes the NBD1:ICL4 interaction, as suggested previously (He et al., 2010; Serohijos et al., 2008) and in Figure S2A. Login to comment
195 Indeed, G550E, besides being able to promote rescue of F508del-CFTR (DeCarvalho et al., 2002), shows the largest combined effect with R1070W (or V510D). Login to comment
197 Moreover, the observed synergy of G550E or R555K with VX-809, but not VRT-325, is consistent with this model in which VRT-325 acts on the NBD1:NBD2 dimerization interface (also supported by the synergy between R1070W and VRT-325). Login to comment
198 Further modeling is, however, required to examine the observation that R1070W is still additive with VX-809 (albeit modestly). Login to comment
210 Our data also show that low temperature, similar to chemical correctors, further increases processing levels of F508del-CFTR by the five genetic revertants, although to variable levels: V510D (by an additional 35%), G550E (65%), R1070W (38%), 4RK (27%), and R555K (22%). Login to comment
225 Assessing the synergistic/additive effects of investigational drug VX-809, one of the most promising to rescue the F508del-CFTR-trafficking defect, with those of genetic revertants as well as other correctors (VRT-325 and Corr-4a) or low temperature pointed to major insights into its MoA: (1) VX-809 is additive to both VRT-325 and Corr-4a, suggesting that each compound operates by a different MoA; (2) VX-809 is additive to low temperature rescue of the mutant almost to WT-CFTR levels; (3) VX-809, VRT-325, and Corr-4a show variable additive effects with the genetic revertants tested (4RK, G550E, and R1070W), thus providing clues for their possible action being exerted at specific protein binding pockets: VX-809 at the NBD1:TMD2 interface (and VRT-325 at NBD1:NBD2) or acting unspecifically (Corr-4a); and (4) VX-809 does not rescue the diacidic code traffic mutant in contrast to low temperature, which seems to act at trafficking surveillance checkpoints. Login to comment
231 EXPERIMENTAL PROCEDURES Cells and Culture Conditions BHK cell lines expressing F508del-4RK (R29K/R516K/R555K/R716K)-, F508del-G550E-, F508del-R1070W-, F508del-V510D-, F508del-R555K-, F508del-V510D/G550E-, F508del-G550E/R1070W-, DAA (D567A)-, 4RK- DAA-, DD/AA (D565A, D567A)-, 4RK-DD/AA-, and R560T-CFTR were produced and cultured as previously described (Roxo-Rosa et al., 2006). Login to comment

PMID: 23891399 [PubMed] Van Goor F et al: "Effect of ivacaftor on CFTR forms with missense mutations associated with defects in protein processing or function."

No. Sentence Comment

44 None M1V A46D E56K P67L R74W G85E E92K D110E D110H R117C R117H E193K L206W R334W I336K T338I S341P R347H R347P R352Q A455E L467P S492F F508del V520F A559T R560S R560T A561E Y569D D579G R668C L927P S945L S977F L997F F1052V H1054D K1060T L1065P R1066C R1066H R1066M A1067T R1070Q R1070W F1074L L1077P H1085R M1101K D1152H S1235R D1270N N1303K 0 100 200 300 400 500 600 * * * CFTR Mutation mRNA (% Normal CFTR) Fig. 1. Login to comment
64 Mutant CFTR form CFTR processing Mature/total % Normal CFTR Normal 0.89 &#b1; 0.01 100.0 &#b1; 18.5 G85E -0.05 &#b1; 0.04 -1.0 &#b1; 0.9 R560S 0.00 &#b1; 0.00 0.0 &#b1; 0.0 R1066C 0.02 &#b1; 0.01 0.0 &#b1; 0.0 S492F 0.00 &#b1; 0.00 0.1 &#b1; 0.1 R560T 0.01 &#b1; 0.01 0.2 &#b1; 0.1 V520F 0.05 &#b1; 0.03 0.3 &#b1; 0.2 M1101K 0.05 &#b1; 0.03 0.3 &#b1; 0.1 A561E 0.08 &#b1; 0.04 0.5 &#b1; 0.2 R1066M 0.02 &#b1; 0.02 0.5 &#b1; 0.4 N1303K 0.02 &#b1; 0.02 0.5 &#b1; 0.3 A559T 0.16 &#b1; 0.09 0.6 &#b1; 0.2 M1V 0.06 &#b1; 0.06 0.7 &#b1; 0.6 Y569D 0.11 &#b1; 0.04 0.6 &#b1; 0.2 R1066H 0.08 &#b1; 0.02a 0.7 &#b1; 0.2a L1065P 0.05 &#b1; 0.05 1.0 &#b1; 0.8 L467P 0.10 &#b1; 0.07 1.2 &#b1; 0.8 L1077P 0.08 &#b1; 0.04 1.5 &#b1; 0.6 A46D 0.21 &#b1; 0.08 1.9 &#b1; 0.5a E92K 0.06 &#b1; 0.05 1.9 &#b1; 1.3 H1054D 0.09 &#b1; 0.04 1.9 &#b1; 0.8 F508del 0.09 &#b1; 0.02a 2.3 &#b1; 0.5a H1085R 0.06 &#b1; 0.01a 3.0 &#b1; 0.7a I336K 0.42 &#b1; 0.05a 6.5 &#b1; 0.7a L206W 0.35 &#b1; 0.10a 6.8 &#b1; 1.7a F1074L 0.52 &#b1; 0.03a 10.9 &#b1; 0.6a A455E 0.26 &#b1; 0.10a 11.5 &#b1; 2.5a E56K 0.29 &#b1; 0.04a 12.2 &#b1; 1.5a R347P 0.48 &#b1; 0.04a 14.6 &#b1; 1.8a R1070W 0.61 &#b1; 0.04a 16.3 &#b1; 0.6a P67L 0.36 &#b1; 0.04a 28.4 &#b1; 6.8a R1070Q 0.90 &#b1; 0.01a 29.5 &#b1; 1.4a S977F 0.97 &#b1; 0.01a 37.3 &#b1; 2.4a A1067T 0.78 &#b1; 0.03a 38.6 &#b1; 6.1a D579G 0.72 &#b1; 0.02a 39.3 &#b1; 3.1a D1270N 1.00 &#b1; 0.00a,c 40.7 &#b1; 1.2a S945L 0.65 &#b1; 0.04a 42.4 &#b1; 8.9a L927P 0.89 &#b1; 0.01a,b 43.5 &#b1; 2.5a,b R117C 0.87 &#b1; 0.02a,b 49.1 &#b1; 2.9a,b T338I 0.93 &#b1; 0.03a,b 54.2 &#b1; 3.7a,b L997F 0.90 &#b1; 0.04a,b 59.8 &#b1; 10.4a,b D110H 0.97 &#b1; 0.01a,b 60.6 &#b1; 1.5a,b S341P 0.79 &#b1; 0.02a 65.0 &#b1; 4.9a,b R668C 0.94 &#b1; 0.03a,b 68.5 &#b1; 1.9a,b R74W 0.78 &#b1; 0.01a 69.0 &#b1; 2.7a,b D110E 0.92 &#b1; 0.05a,b 87.5 &#b1; 9.5a,b R334W 0.91 &#b1; 0.05a,b 97.6 &#b1; 10.0a,b K1060T 0.87 &#b1; 0.02a,b 109.9 &#b1; 28.0a,b R347H 0.96 &#b1; 0.02a,c 120.7 &#b1; 2.8a,b S1235R 0.96 &#b1; 0.00a,c 139.0 &#b1; 9.0a,b E193K 0.84 &#b1; 0.02a,b 143.0 &#b1; 17.1a,b R117H 0.86 &#b1; 0.01a,b 164.5 &#b1; 34.2a,b R352Q 0.98 &#b1; 0.01a,b 179.9 &#b1; 8.0a,c F1052V 0.90 &#b1; 0.01a,b 189.9 &#b1; 33.1a,b D1152H 0.96 &#b1; 0.02a,c 312.0 &#b1; 45.5a,b Notes to Table 1: Quantification of steady-state CFTR maturation expressed as the mean (&#b1;SEM; n = 5-9) ratio of mature CFTR to total CFTR (immature plus mature) or level of mature mutant CFTR relative to mature normal-CFTR (% normal CFTR) in FRT cells individually expressing CFTR mutations. Login to comment
74 Because the level of CFTR mRNA was similar across the panel of cell lines tested, the range in baseline activity and ivacaftor response likely reflects the severity of the functional defect and/or the 0 50 100 150 200 S341P R347P L467P S492F A559T A561E Y569D L1065P R1066C R1066M L1077P M1101K N1303K R560S L927P R560T H1085R V520F E92K M1V F508del H1054D I336K A46D G85E R334W T338I R1066H R352Q R117C L206W R347H S977F S945L A455E F1074L E56K P67L R1070W D110H D579G D110E R1070Q L997F A1067T E193K R117H R74W K1060T R668C D1270N D1152H S1235R F1052V Baseline With ivacaftor * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * Chloride transport (% Normal) Mutant CFTR form 0 100 200 300 400 S341P R347P L467P S492F A559T A561E Y569D L1065P R1066C R1066M L1077P M1101K N1303K R560S L927P R560T H1085R V520F E92K M1V F508del H1054D I336K A46D G85E R334W T338I R1066H R352Q R117C L206W R347H S977F S945L A455E F1074L P67L E56K R1070W D110H D579G D110E R1070Q L997F A1067T E193K R117H R74W K1060T R668C D1270N D1152H S1235R F1052V * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * Mature CFTR (% Normal) Mutant CFTR form A B Fig. 2. Login to comment
82 Mutation Patientsa Chloride transport (bc;A/cm2 ) Chloride transport (% normal) EC50 Baseline With ivacaftor Baseline With ivacaftor Fold increase over baselineb Normal 204.5 &#b1; 33.3 301.3 &#b1; 33.8c 100.0 &#b1; 16.3 147.3 &#b1; 16.5c 1.5 266 &#b1; 42 G551D 1282 1.5 &#b1; 0.7 113.2 &#b1; 13.0c 1.0 &#b1; 0.5 55.3 &#b1; 6.3c 55.3 312 &#b1; 73 F1052V 12 177.3 &#b1; 13.7 410.2 &#b1; 11.3c 86.7 &#b1; 6.7 200.7 &#b1; 5.6c 2.3 177 &#b1; 14 S1235R ND 160.6 &#b1; 25.7 352.1 &#b1; 43.4c 78.5 &#b1; 12.6 172.2 &#b1; 21.2c 2.2 282 &#b1; 104 D1152H 185 117.3 &#b1; 23.0 282.7 &#b1; 46.9c 57.4 &#b1; 11.2 138.2 &#b1; 22.9c 2.4 178 &#b1; 67 D1270N 32 109.5 &#b1; 20.5 209.5 &#b1; 27.4c 53.6 &#b1; 10.0 102.4 &#b1; 13.4c 1.9 254 &#b1; 56 R668C 45 99.0 &#b1; 9.4 217.6 &#b1; 11.7c 48.4 &#b1; 4.6 106.4 &#b1; 5.7c 2.2 517 &#b1; 105 K1060T ND 89.0 &#b1; 9.8 236.4 &#b1; 20.3c 43.5 &#b1; 4.8 115.6 &#b1; 9.9c 2.7 131 &#b1; 73 R74W 25 86.8 &#b1; 26.9 199.1 &#b1; 16.8c 42.5 &#b1; 13.2 97.3 &#b1; 8.2c 2.3 162 &#b1; 17 R117H 739 67.2 &#b1; 13.3 274.1 &#b1; 32.2c 32.9 &#b1; 6.5 134.0 &#b1; 15.7c 4.1 151 &#b1; 14 E193K ND 62.2 &#b1; 9.8 379.1 &#b1; 1.1c 30.4 &#b1; 4.8 185.4 &#b1; 1.0c 6.1 240 &#b1; 20 A1067T ND 55.9 &#b1; 3.2 164.0 &#b1; 9.7c 27.3 &#b1; 1.6 80.2 &#b1; 4.7c 2.9 317 &#b1; 214 L997F 27 43.7 &#b1; 3.2 145.5 &#b1; 4.0c 21.4 &#b1; 1.6 71.2 &#b1; 2.0c 3.3 162 &#b1; 12 R1070Q 15 42.0 &#b1; 0.8 67.3 &#b1; 2.9c 20.6 &#b1; 0.4 32.9 &#b1; 1.4c 1.6 164 &#b1; 20 D110E ND 23.3 &#b1; 4.7 96.4 &#b1; 15.6c 11.4 &#b1; 2.3 47.1 &#b1; 7.6c 4.1 213 &#b1; 51 D579G 21 21.5 &#b1; 4.1 192.0 &#b1; 18.5c 10.5 &#b1; 2.0 93.9 &#b1; 9.0c 8.9 239 &#b1; 48 D110H 30 18.5 &#b1; 2.2 116.7 &#b1; 11.3c 9.1 &#b1; 1.1 57.1 &#b1; 5.5c 6.2 249 &#b1; 59 R1070W 13 16.6 &#b1; 2.6 102.1 &#b1; 3.1c 8.1 &#b1; 1.3 49.9 &#b1; 1.5c 6.2 158 &#b1; 48 P67L 53 16.0 &#b1; 6.7 88.7 &#b1; 15.7c 7.8 &#b1; 3.3 43.4 &#b1; 7.7c 5.6 195 &#b1; 40 E56K ND 15.8 &#b1; 3.1 63.6 &#b1; 4.4c 7.7 &#b1; 1.5 31.1 &#b1; 2.2c 4.0 123 &#b1; 33 F1074L ND 14.0 &#b1; 3.4 43.5 &#b1; 5.4c 6.9 &#b1; 1.6 21.3 &#b1; 2.6c 3.1 141 &#b1; 19 A455E 120 12.9 &#b1; 2.6 36.4 &#b1; 2.5c 6.3 &#b1; 1.2 17.8 &#b1; 1.2c 2.8 170 &#b1; 44 S945L 63 12.3 &#b1; 3.9 154.9 &#b1; 47.6c 6.0 &#b1; 1.9 75.8 &#b1; 23.3c 12.6 181 &#b1; 36 S977F 9 11.3 &#b1; 6.2 42.5 &#b1; 19.1c 5.5 &#b1; 3.0 20.8 &#b1; 9.3c 3.8 283 &#b1; 36 R347H 65 10.9 &#b1; 3.3 106.3 &#b1; 7.6c 5.3 &#b1; 1.6 52.0 &#b1; 3.7c 9.8 280 &#b1; 35 L206W 81 10.3 &#b1; 1.7 36.4 &#b1; 2.8c 5.0 &#b1; 0.8 17.8 &#b1; 1.4c 3.6 101 &#b1; 13 R117C 61 5.8 &#b1; 1.5 33.7 &#b1; 7.8c 2.9 &#b1; 0.7 16.5 &#b1; 3.8c 5.7 380 &#b1; 136 R352Q 46 5.5 &#b1; 1.0 84.5 &#b1; 7.8c 2.7 &#b1; 0.5 41.3 &#b1; 3.8c 15.2 287 &#b1; 75 R1066H 29 3.0 &#b1; 0.3 8.0 &#b1; 0.8c 1.5 &#b1; 0.1 3.9 &#b1; 0.4c 2.6 390 &#b1; 179 T338I 54 2.9 &#b1; 0.8 16.1 &#b1; 2.4c 1.4 &#b1; 0.4 7.9 &#b1; 1.2c 5.6 334 &#b1; 38 R334W 150 2.6 &#b1; 0.5 10.0 &#b1; 1.4c 1.3 &#b1; 0.2 4.9 &#b1; 0.7c 3.8 259 &#b1; 103 G85E 262 1.6 &#b1; 1.0 1.5 &#b1; 1.2 0.8 &#b1; 0.5 0.7 &#b1; 0.6 NS NS A46D ND 2.0 &#b1; 0.6 1.1 &#b1; 1.1 1.0 &#b1; 0.3 0.5 &#b1; 0.6 NS NS I336K 29 1.8 &#b1; 0.2 7.4 &#b1; 0.1c 0.9 &#b1; 0.1 3.6 &#b1; 0.1c 4 735 &#b1; 204 H1054D ND 1.7 &#b1; 0.3 8.7 &#b1; 0.3c 0.8 &#b1; 0.1 4.2 &#b1; 0.1c 5.3 187 &#b1; 20 F508del 29,018 0.8 &#b1; 0.6 12.1 &#b1; 1.7c 0.4 &#b1; 0.3 5.9 &#b1; 0.8c 14.8 129 &#b1; 38 M1V 9 0.7 &#b1; 1.4 6.5 &#b1; 1.9c 0.4 &#b1; 0.7 3.2 &#b1; 0.9c 8.0 183 &#b1; 85 E92K 14 0.6 &#b1; 0.2 4.3 &#b1; 0.8c 0.3 &#b1; 0.1 2.1 &#b1; 0.4c 7.0 198 &#b1; 46 V520F 58 0.4 &#b1; 0.2 0.5 &#b1; 0.2 0.2 &#b1; 0.1 0.2 &#b1; 0.1 NS NS H1085R ND 0.3 &#b1; 0.2 2.1 &#b1; 0.4 0.2 &#b1; 0.1 1.0 &#b1; 0.2 NS NS R560T 180 0.3 &#b1; 0.3 0.5 &#b1; 0.5 0.1 &#b1; 0.1 0.2 &#b1; 0.2 NS NS L927P 15 0.2 &#b1; 0.1 10.7 &#b1; 1.7c 0.1 &#b1; 0.1 5.2 &#b1; 0.8c 52.0 313 &#b1; 66 R560S ND 0.0 &#b1; 0.1 -0.2 &#b1; 0.2 0.0 &#b1; 0.0 -0.1 &#b1; 0.1 NS NS N1303K 1161 0.0 &#b1; 0.0 1.7 &#b1; 0.3 0.0 &#b1; 0.0 0.8 &#b1; 0.2 NS NS M1101K 79 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS L1077P 42 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS R1066M ND 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS R1066C 100 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS L1065P 25 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS Y569D 9 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS A561E ND 0.0 &#b1; 0.1 0.0 &#b1; 0.1 0.0 &#b1; 0.0 0.0 &#b1; 0.1 NS NS A559T 43 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS S492F 16 0.0 &#b1; 0.0 1.7 &#b1; 1.2 0.0 &#b1; 0.0 0.8 &#b1; 0.6 NS NS L467P 16 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS R347P 214 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS S341P 9 0.0 &#b1; 0.0 0.2 &#b1; 0.2 0.0 &#b1; 0.0 0.1 &#b1; 0.1 NS NS a Number of individuals with the individual mutation in the CFTR-2 database (www.CFTR2.org). Login to comment
86 For example, the baseline level of chloride transport and ivacaftor response was higher for mutant CFTR forms associated with mild defects in CFTR processing (e.g., E56K, P67L, L206W, A455E, D579G, S945L, S977F, A1067T, R1070Q, R1070W, F1074L, and D1270N) than for those associated with severe defects in CFTR processing (e.g., F508del, H1054D, R1066H). Login to comment
89 For mutant CFTR forms that have multiple defects (e.g., R117H, F508del, S945L, R1070Q, A1067T, R1070W, and R347P), the relative impact of each defect is likely to affect the magnitude of the baseline chloride transport and ivacaftor response in vitro and in a clinical setting. Login to comment
92 Mutant CFTR forms that did not significantly respond to ivacaftor under the experimental conditions used in this study were generally associated with severe defects in CFTR processing A B C D E F 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 S1235R D1152H F1052V D1270N ivacaftor [Log M] 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 R668C K1060T R74W R117H ivacaftor [Log M] 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 E193K A1067T L997F R1070Q ivacaftor [Log M] Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 D110E D579G D110H R1070W ivacaftor [Log M] 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 F1074L E56K P67L A455E ivacaftor [Log M] 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 R347H S945L L206W S977F ivacaftor [Log M] 0 100 200 300 400 -8 -6 -4 0 T338I R1066H R117C R352Q ivacaftor [Log M] 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 F508del R334W H1054D E92K ivacaftor [Log M] 0 5 10 15 20 -9 -8 -7 -6 -5 -4 0 F508del R334W H1054D E92K R1066H T338I ivacaftor [Log M] G H I Fig. 3. Login to comment

PMID: 23974870 [PubMed] Sosnay PR et al: "Defining the disease liability of variants in the cystic fibrosis transmembrane conductance regulator gene."

No. Sentence Comment

137 In addition to these ten variants, c.1210-12(7) (legacy name 7T) had already been reported to be non-penetrant48 and was identified as a second variant in numerous fathers, and a twelfth variant, p.Ile1027Thr, was deemed 159 variants ࣙ0.01% frequency in CFTR2 127 variants meet clinical and functional criteria Clinical and functional analysis 13 variants meet neither criteria 14 variants 5 variants 7 variants 6 variants Evidence of non-penetrance No evidence of non-penetrance 19 variants meet clinical or functional criteria 127 variants are CF causing 12 variants are non CF causing 20 variants are indeterminate p.Arg117HisߤC p.Arg75Gln p.Gly576Alaߤ p.Arg668Cys ߤ p.Met470Val C p.IIe1027Thr ߤC p.Val754Met ߤC p.IIe148Thr ߤC p.Arg31Cys C p.Ser1235Arg ߤ p.Leu997Phe ߤ p.Arg1162Leu p.Leu227Arg F p.Gln525* F p.Leu558SerC p.Asp614Gly C c.2657+2_2657+3insA C c.1418delG F c.1210-12(7) ߤ p.Arg1070Gln C p.Asp1270Asn ߤC p.[Gln359Lys; Thr360Lys] p.Gly1069Argߤ p.Asp1152His p.Phe1052Val c.1210-12(5) p.Arg74Trpߤ p.IIe1234Val ߤC p.Arg1070Trp ߤF p.Ser977Phe F p.Asp579Gly C p.Tyr569Asp F Penetrance analysis Figure 4ߒ Assignment of disease liability to the 159 most frequent CFTR variants using three criteria. Login to comment

PMID: 24412276 [PubMed] Loo TW et al: "The cystic fibrosis V232D mutation inhibits CFTR maturation by disrupting a hydrophobic pocket rather than formation of aberrant interhelical hydrogen bonds."

No. Sentence Comment

169 For example, V510D promotes maturation of mutants with processing mutations in TMD1 (V232D), TMD2 (H1085R) and NBD1 (DF508) whereas other suppressors such as I539T and R1070W promote maturation of DF508 CFTR but not mutants V232D or H1085R [19]. Login to comment

PMID: 24685677 [PubMed] Pranke IM et al: "Biosynthesis of cystic fibrosis transmembrane conductance regulator."

No. Sentence Comment

1442 Mutations located in NBD1, such as I539T, G550E, R553M/Q and R555K, as well as R1070W in CL4 of MSD2 promote Phe508del-CFTR maturation and trafficking to the cell surface and also restore channel activity (DeCarvalho et al., 2002; Teem et al., 1993, 1996; Thibodeau et al., 2010). Login to comment

PMID: 24726831 [PubMed] Eckford PD et al: "VX-809 and related corrector compounds exhibit secondary activity stabilizing active F508del-CFTR after its partial rescue to the cell surface."

No. Sentence Comment

25 To date, however, there have been conflicting reports regarding the consequences of mutations at the NBD1-ICL4 interface (including R1070W) on the efficacy of VX-809 in in-cell maturation studies (He et al., 2013; Okiyoneda et al., 2013), prompting speculation that VX-809 may not bind directly to this site on the mutant protein. Login to comment

PMID: 24737137 [PubMed] Phuan PW et al: "Synergy-based small-molecule screen using a human lung epithelial cell line yields DeltaF508-CFTR correctors that augment VX-809 maximal efficacy."

No. Sentence Comment

2 The interfacial stability defect can be partially corrected by the investigational drug VX-809 (3-[6-[[[1-(2,2- difluoro-1,3-benzodioxol-5-yl)cyclopropyl]carbonyl]amino]-3-methyl-2-pyridinyl]-benzoic acid) or the R1070W mutation. Login to comment
4 We postulated that a second corrector targeting a distinct folding/interfacial defect might act in synergy with VX-809 or the R1070W suppressor mutation. Login to comment
5 A biochemical screen for DF508-CFTR cell surface expression was developed in a human lung epithelium-derived cell line (CFBE41o2 ) by expressing chimeric CFTRs with a horseradish peroxidase (HRP) in the fourth exofacial loop in either the presence or absence of R1070W. Login to comment
35 Therefore, novel corrector molecules targeting either the NBD1 and/or its interface defects should be preferentially identified by screening in the background of VX-809 or the interface-stabilizing mutation R1070W. Login to comment
43 The cloning and characterization of 3HA-tagged variants of ƊF508-CFTR, R1070W- ƊF508-CFTR, and 3S-ƊF508-CFTR (containing the F494N, Q637R, and F429S NBD1 suppressor mutations) were described (Okiyoneda et al., 2013). Login to comment
58 In one set of assays, R1070W-ƊF508-CFTR-HRP (R1070W-HRP)-expressing CFBE41o2 cells were incubated with 100 ml medium containing 25 mM test compounds and 0.5 mg/ml doxycycline for 24 hours at 37&#b0;C. Login to comment
87 A second screen ("R1070W screen") (Fig. 1B) used CFBE41o2 cells transfected with DF508-CFTR-HRP containing a R1070W mutation (R1070W-HRP CFBE41o2 ). Login to comment
90 For the ƊF508 screen, cells were incubated with test compounds (at 25 mM) together with 2 mM VX-809; for the R1070W screen, cells were incubated with test compounds (at 25 mM) alone. Login to comment
96 Similarly, with the R1070W-HRP CFBE41o2 cells, VX-809 increased the signal maximally to approximately 220 a.u. over the DMSO control baseline of approximately 85 a.u., representing an approximately 2.5-fold signal increase (bar graphs in Fig. 1). Login to comment
98 Low-temperature rescue (27&#b0;C) of ƊF508-CFTR increased the HRP luminescence signal by approximately 2-fold (compared with 37&#b0;C) in ƊF508-HRP CFBE41o2 cells and approximately 3-fold in R1070W-HRP CFBE41o2 cells. Login to comment
100 EC50 values were 30 and 78 nM in the low temperature-rescued ƊF508-HRP and R1070W-HRP CFBE41o2 cells, respectively. Login to comment
101 Preferential correction of DF508-CFTR-3HA with the NBD1 stabilizing 3S mutations (F494N, Q637R, and F429S) compared with CFTR carrying the R1070W interface-stabilizing mutation has been taken as evidence that VX-809 preferentially stabilizes the interface between NBD1 and MSDs but not the NBD1 folding defect CFTR (Okiyoneda et al., 2013). Login to comment
103 The relative insensitivity of R1070W-HRP to VX-809 was used to identify correctors that act in synergy with VX-809. Login to comment
105 A total of 110,240 drug-like small synthetic molecules were tested in the ƊF508 and R1070W screens. Login to comment
108 For the R1070W screen, 25 active compounds were identified based on a .50% increase in the luminescence signal over that of DMSO. Login to comment
109 After retesting, nine compounds, grouped into six classes, were confirmed from the R1070W screen. Figure 2D shows structures of the six most active compounds (A-01, B-01, C-01, D-01, E-01, and F-01). Login to comment
110 Because different small-molecule collections were used for the ƊF508 and R1070W screens, we cross-tested all active correctors in both the ƊF508-HRP and R1070W-HRP CFBE41o2 cell lines (Supplemental Fig. 2). Login to comment
112 However, compounds D-01, E-01, and F-01, discovered from the R1070W screen, were not active in ƊF508-HRP CFBE41o2 cells. Login to comment
113 J-01, discovered from the ƊF508 screen, was not active in R1070W-HRP CFBE41o2 cells. Login to comment
117 We also measured the concentration-dependent activities of A-01, B-01, D-01, H-01, and K-01 (Fig. 3B) in R1070W-CFBE41o2 cells and found that D-01 is the most potent corrector, with an EC50 value of approximately 1.2 mM and a maximal signal that is approximately 65% of that produced by 2 mM VX-809. Login to comment
126 Similar compound potency and efficacy were found for class D analogs in R1070W-HRP CFBE41o2 cells (Fig. 4C). Login to comment
141 (B) Screening assay for R1070W-ƊF508-CFTR-HRP CFBE41o cells (top) showing incubation with 25 mM test compounds for 24 hours at 37&#b0;C. Login to comment
163 (C) Summary of primary findings of the R1070W-DF508-CFTR screen. Login to comment
170 To investigate whether any of the correctors identified here may target the NBD2 interface, their synergy with C4 was evaluated in R1070W-HRP CFBE41o2 cells using the HRP luminescence assay. Login to comment
191 (B) Concentration-dependence data of A-01, B-01, D-01, H-01, and K-01 in R1070W-ƊF508-CFTR-HRP CFBE41o2 cells (S.E., n = 3). Login to comment
197 (A) Concentration-dependence data of class D analogs in R1070W-ƊF508-CFTR-HRP CFBE41o2 cells (S.E., n = 3). Login to comment
212 A second screen was done in cells expressing R1070W-ƊF508-CFTR (in the absence of VX-809), since in the background of genetically stabilized ƊF508-NBD1, the R1070W mutation was necessary and sufficient to restore robust CFTR domain assembly and cell surface expression (Thibodeau et al., 2010; Mendoza et al., 2012; Rabeh et al., 2012). Login to comment
213 Screening was performed using a human lung epithelium-derived cell line (CFBE41o2 ) that was stably transfected with HRP-tagged ƊF508-CFTR or R1070W-ƊF508-CFTR. Login to comment
227 (C) Relative effect of corrector 4 (10 mM, 24 hours, 37&#b0;C) on the PM density of R1070W-HRP CFBE41o2 cells treated with A-01, B-01, C-01, D-01, H-01, J-01, K-01, or VX-809 at the indicated concentrations. Login to comment

PMID: 24949105 [PubMed] Cebotaru L et al: "Complement yourself: Transcomplementation rescues partially folded mutant proteins."

No. Sentence Comment

89 Recent experiments have shown that certain amino acid substitutions, namely R1070W in combination with ƊF508 CFTR, can rescue ƊF508 CFTR trafficking, again pointing to the importance of domain-domain interactions in the mis-processing of ƊF508 (62). Login to comment

PMID: 25083918 [PubMed] He L et al: "Restoration of NBD1 thermal stability is necessary and sufficient to correct F508 CFTR folding and assembly."

No. Sentence Comment

30 Furthermore, the combination of maximal NBD1 stabilization and interface modification by the paradoxical R1070W interface mutation disrupts the normal control of channel activity by phosphorylation. Login to comment
35 Rabeh et al. found that, while several of the solubilizing and suppressor mutations caused only a modest promotion of ƊF508 CFTR maturation, their effects were greater when they were combined with the NBD1/CL4 interface substitution R1070W or V510D [33], the latter also having been shown to have a direct effect on NBD1 thermostability [10]. Login to comment
55 In contrast, each of the interface substitutions, V510D (lane 8) and R1070W (lane 9), had much smaller effects. Login to comment
61 The rates of appearance of the mature products and the levels reached were increased further when the R1070W mutation was added to the combined NBD1 changes but not when V510D was added. Login to comment
84 The surface level of ƊF508 CFTR with the combined NBD1 stabilizing mutations (ƊF/combo) reached ~90% that of WT CFTR (Fig. 1c), and the level was somewhat further increased when V510D or R1070W was added to the combination (ƊF/combo/V510D and ƊF/combo/R1070W). Login to comment
87 Functional activity measured using this iodide efflux assay was similar for the ƊF508/combo and the ƊF508/4PT/R1070W, indicating the effectiveness of the combination of multiple NBD1 stabilizing mutations without interface modification in promoting ƊF508 CFTR channel activity. Login to comment
89 It was initially surprising to observe that the addition of R1070W to the ƊF508/combo apparently greatly diminished iodide efflux (Fig. 1d, bottom panel). Login to comment
91 This apparent inability of baby hamster kidney (BHK) cells expressing ƊF508/combo/R1070W CFTR to be effectively stimulated by forskolin cocktail suggested that the ƊF508/combo/R1070W CFTR channel might be constitutively active, a possibility that was subsequently confirmed at the single-channel level (see Fig. 5 below). Login to comment
111 The R1070W mutation in CL4 has different effects on the gating of WT and ƊF508 CFTR channels. Login to comment
112 (a) Western blot indicating enhancement of maturation of mutant R1070W and ƊF508 CFTRs by combined NBD1 stabilizing mutants (combo). Login to comment
113 (b and c) Single-channel recordings of WT (b) and ƊF508 CFTR (c) containing the R1070W mutant at 25 &#b0;C and 35 &#b0;C. All point histograms used to calculate single-channel parameters are shown to the left of each tracing. Login to comment
116 (d) WT and ƊF508/R1070W CFTR single-channel recordings during continuous temperature ramp of 1 &#b0;C/min from 30 to 35 &#b0;C, maintained at 30 &#b0;C for 3 min before initiating the ramp and held at 35 &#b0;C for 2 min. Login to comment
117 The moment of ƊF508/R1070W functional inactivation is shown by an arrow above the tracing and appeared after about 1 min at 35 &#b0;C. X-scale bar below trace is 60 s and Y-scale bar is 1 pA. 1.5 mM, reflecting low-affinity binding. Login to comment
126 The BEIA influence on the other interface substituted form of the protein, R1070W, was less pronounced, particularly at the lower concentrations of the compound. Login to comment
129 The disease-associated CL4 mutation R1070W differentially influences WT and ƊF508 CFTR R1070W is a relatively rare disease-associated mutation in the CL4 region of CFTR. Login to comment
131 Homology models of CFTR three-dimensional structure indicated that the R1070-containing segment of CL4 was proximal to F508 on the NBD1 surface [29-32] and Thomas et al. reasoned that the introduction of the aromatic tryptophan side chain by R1070W substitution might compensate for the loss of the phenylalanine side chain due to the absence of F508 at the interface between domains [8]. Login to comment
132 They showed that the R1070W mutation substantially improved the maturation of ƊF508 CFTR. Login to comment
135 Combined NBD1 stabilizing mutations promote maturation and function of R1070W CFTR in both ƊF508 and WT backgrounds. Login to comment
136 (a) Single-channel current tracings of the WT, ƊF508/Combo and ƊF508/Combo/R1070W CFTRs at 25 &#b0;C. All point histograms, single-channel conductances and open probabilities are shown as in Fig. 4. Login to comment
137 (b) Single-channel currents at 37 &#b0;C showing the influence of combined NBD1 stabilizing changes on ƊF508 and WT CFTR without and with the R1070W mutation. Login to comment
139 Combined NBD1 stabilizing mutations and R1070W result in phosphorylation-independent opening of ƊF508 CFTR channels. Login to comment
140 (a) Single-channel recordings at 37 &#b0;C of alkaline phosphatase treated ƊF508 CFTR containing combined NBD1 stabilizing mutations without (upper tracing) and with R1070W (middle tracing) as indicated. Login to comment
141 Lower tracing: single-channel recording at 37 &#b0;C of ƊF508/combo/R1070W with all 15 PKA phosphorylation sites mutated (15SA). Login to comment
145 Contrasting effects of R1070W on WT and ƊF508 CFTR is very evident at the level of biosynthetic processing as shown in Fig. 4a, where maturation of the WT is reduced while that of ƊF508 is promoted. Login to comment
148 While the opposite effects of R1070W on WT and ƊF508 CFTR maturation have been well documented, the corresponding functional consequences have not. Login to comment
150 The R1070W mutation alone resulted in very low channel open probabilities at both 25 &#b0;C and 35 &#b0;C (Fig. 4b). Login to comment
153 In contrast, robust channel gating was observed with the combined ƊF508/ R1070W variant with the expected elevation of Po as temperature was increased from 25 &#b0;C to 35 &#b0;C (Fig. 4c). Login to comment
156 Therefore, as with the much less active R1070W/WT form, this more active double mutant (ƊF508/R1070W) has a similar short functional lifetime at 35 &#b0;C confirming that it remains thermally unstable [15]. Login to comment
158 This behavior was especially evident when the single-channel activities of ƊF508 CFTR modified by combined NBD1 stabilizing changes with and without the R1070W interface mutation were compared with the unmodified WT at both 25 &#b0;C (Fig. 5a) and 37 &#b0;C (Fig. 5b). Login to comment
160 Strikingly, addition of the R1070W mutation to the NBD1 stabilized combination resulted in a very high open probability (0.91) even at this lower temperature. Login to comment
162 With the addition of R1070W, a nearly, completely locked open state was achieved with a Po of 0.98 (Fig. 5b, third tracing). Login to comment
164 The lower two tracings in Fig. 5b indicate that the combined stabilizing modifications of NBD1 also elevate the open probability of the WT channel with and without the R1070W mutation. Login to comment
165 Combined NBD1 stabilization and NBD1/CL4 interface modification result in constitutive ion channel activity of ƊF508 CFTR In iodide efflux experiments from whole cells expressing ƊF508/combo/R1070W CFTR, we had observed that the cells could not be efficiently loaded with the anion prior to the stimulation of cAMP production to activate protein kinase A (PKA) (Fig. 1d), suggesting the possibility that the apparent locked-open state might exist in the absence of the normally obligatory phosphorylation by PKA. Login to comment
168 In contrast, the NBD1 combo stabilized versions of the ƊF508 CFTR channels (Fig. 6a, middle tracing) and WT (Fig. 6b, middle tracing) responded very differently to addition of the R1070W mutation, the former channel becoming essentially locked open while the open probability of the latter was increased only modestly. Login to comment
169 To further verify that the high open probability state of ƊF508/combo/R1070W CFTR was reached independently of phosphorylation by PKA, we employed a construct in which serines or threonines at all 15 PKA phosphorylation sites were mutated to Ala (15SA) [39]. Login to comment
170 While the activity of the combo/ R1070W/WT CFTR was little changed by the 15SA modifications (Fig. 6b, lower tracing), the 15SA version of the ƊF508/combo/R1070W CFTR channel remained fully active even in the absence of PKA phosphorylation sites (Fig. 6a, lower tracing), confirming that the maximally NBD1 stabilized and NBD1/CL4 interface modified form of ƊF508 CFTR has lost normal regulatory control by phosphorylation. Login to comment
176 To further pursue this hypothesis here, we combined several of the most effective second-site changes in NBD1 with or without the interface stabilizing mutant R1070W and compared their effects on maturation, traffic to the cell surface and channel function. Login to comment
200 Turning to the impact of modifying the NBD1/CL4 interface, which may be a site of action of the VX-809 corrector [23,25], the R1070W mutation, shown by others to improve ƊF508 CFTR maturation [8], was the main tool we employed. Login to comment
202 Not only were their effects small compared to those of NBD1 stabilizing changes but R1070W also caused only a small increment above the correction caused by combined NBD1 stabilization (Fig. 1). Login to comment
203 From a functional perspective, most significant was the finding that, when modified by combined NBD1 mutations and R1070W, the efficiently assembled ƊF508 channel was constitutively active, not requiring phosphorylation by PKA (Fig. 6). Login to comment
204 Although the mechanism underlying this effect remains to be elucidated, it occurs in the stabilized mutant protein only when the NBD1/CL4 interface is altered by both the absence of F508 and the presence of R1070W (Figs. Login to comment
206 The disease-associated R1070W mutation alone is destabilizing of WT CFTR but does not cause it to be constitutively active without phosphorylation even when it contains all the NBD1 stabilizing changes (Fig. 6). Login to comment
208 These findings do not detract from the idea of therapeutically targeting the interface in different ways since it clearly plays a role in the propagation of stabilizing effects between domains as evidenced by the fact that impaired maturation caused by R1070W can be overcome by NBD1 stabilizing changes (Fig. 4a). Login to comment

PMID: 25287046 [PubMed] Mornon JP et al: "Full-open and closed CFTR channels, with lateral tunnels from the cytoplasm and an alternative position of the F508 region, as revealed by molecular dynamics."

No. Sentence Comment

358 Some other CFTR mutations of varying clinical consequences, such as F1052 V, G1069R, and R1070W/R1070Q complete the list in this region. Login to comment

PMID: 25404111 [PubMed] Cutting GR et al: "Cystic fibrosis genetics: from molecular understanding to clinical application."

No. Sentence Comment

55 Intriguingly, introduction of the disease-associated p.Arg1070Trp (legacy R1070W)29 variant in CL4 and correction of NBD1 misfolding using synthetic suppressor mutations could restore processing to F508delߛCFTR28,30,31 . Login to comment

PMID: 25674778 [PubMed] Baker MW et al: "Improving newborn screening for cystic fibrosis using next-generation sequencing technology: a technical feasibility study."

No. Sentence Comment

31 Both methods used 5 &#b5;l of isolated DNA for the NGS assay. NGS assay for detection of CFTR mutations/variants CFTR mutations are described using both the international nomenclature of the Human Genome Variation Society Mutations that have varying consequences c.3454G>C (D1152H) c.3154T>G (F1052V) c.3208C>T (R1070W) c.2930C>T (S977F) - c.3808G>A (D1270N) c.3205G>A (G1069R) c.350G>A (R117H) PolyTG/ polyT - c.1736A>G (D579G) c.3209G>A (R1070Q) c.220C>T (R74W) - - Mutations still under evaluation c.2657ߙ+ߙ2_2657ߙ+ߙ3insA (2789ߙ+ߙ2insA) c.680T>G (L227R) c.1705T>G (Y569D) - - c.1841A>G (D614G) c.1673T>C (L558S) - - - c.3700A>G (I1234V) c. Login to comment

PMID: 26014425 [PubMed] Girardet A et al: "The improvement of the best practice guidelines for preimplantation genetic diagnosis of cystic fibrosis: toward an international consensus."

No. Sentence Comment

87 [Gln359Lys; Thr360Lys] L558S c.1673 T4C p.Leu558Ser Y569D c.1705 T4G p.Tyr569Asp D579G c.1736 A4G p.Asp579Gly D614G c.1841 A4G p.Asp614Gly S977F c.2930C4T p.Ser977Phe F1052V c.3154 T4G p.Phe1052Val G1069R c.3205G4A p.Gly1069Arg R1070Q c.3209G4A p.Arg1070Gln D1152H c.3454G4C p.Asp1152His I1234V c.3700 A4G p.Ile1234Val 5T c.1210 - 12[5] Examples of common not CF-causing variantsc R31C c.91C4T p.Arg31Cys R74W c.220C4T p.Arg74Trp R75Q c.224G4A p.Arg75Gln I148T c.443 T4C p.Ile148Thr M470V c.1408 A4G p.Met470Val G576A c.1727G4C p.Gly576Ala R668C c.2002C4T p.Arg668Cys V754M c.2260G4A p.Val754Met L997F c.2991G4C p.Leu997Phe I1027T c.3080 T4C p.Ile1027Thr R1070W c.3208C4T p.Arg1070Trp R1162L c.3485G4T p.Arg1162Leu Table 1 (Continued) HGVS nomenclature Legacy name cDNA nucleotide name Protein name S1235R c.3705 T4G p.Ser1235Arg D1270N c.3808G4A p.Asp1270Asn 7T c.1210-12[7] Abbreviation: HGVS, Human Genome Variation Society. Login to comment
99 Missense variants R74W, R1070W, D1270N are classified as 'indeterminate` by Sosnay et al.,17 however, as they are frequently found in trans with a severe CF variant in asymptomatic individuals (including fertile fathers), they may not be sufficient to cause disease.19 Moreover, they are often associated within the same allele (eg in cis), forming various combinations ('complex alleles`) depending on individuals, so that their disease liability is questionable. Login to comment