ABCMdb

PMID: 9482579

Malone G, Haworth A, Schwarz MJ, Cuppens H, Super M
Detection of five novel mutations of the cystic fibrosis transmembrane regulator (CFTR) gene in Pakistani patients with cystic fibrosis: Y569D, Q98X, 296+12(T>C), 1161delC and 621+2(T>C).
Hum Mutat. 1998;11(2):152-7., [PubMed]

Sentences

No. Mutations Sentence Comment

0 ABCC7 p.Tyr569Asp ABCC7 p.Gln98* (c) 1998 WILEY-LISS, INC. RESEARCH ARTICLE Detection of Five Novel Mutations of the Cystic Fibrosis Transmembrane Regulator (CFTR) Gene in Pakistani Patients With Cystic Fibrosis: Y569D, Q98X, 296+12(T>C), 1161delC and 621+2(T>C) Geraldine Malone,1* Andrea Haworth,1 Martin J. Schwarz,1 Harry Cuppens,2 and Maurice Super1 1 North West Regional Molecular Genetics Laboratory, Royal Manchester Children`s Hospital, Manchester, U.K. 2 Centre for Human Genetics, University of Leuven, Leuven, Belgium Communicated by Michel Goossens We analysed DNA samples from 26 Pakistani patients with cystic fibrosis (CF) living in the United Kingdom (14 from patients residing in the north west of England, who were referred directly to the North West Regional Molecular Genetics Laboratory, and 12 from other regional molecular genetics laboratories). Login to comment 1 ABCC7 p.Arg709* Of 56 mutations seen in native U.K. CF patients, only DeltaF508, R709X, and 2184insA were detected in the Pakistani patients. Login to comment 2 ABCC7 p.Tyr569Asp ABCC7 p.Gln98* Combined SSCP/Heteroduplex analysis, DGGE, and direct DNA cycle sequencing revealed five novel mutations: Y569D, Q98X, 296+12(T>C), 1161delC, and 621+2(T>C), which appear to be specific to Pakistani CF families. Login to comment 3 ABCC7 p.Arg560Ser In addition, a novel polymorphism, 297-67(A/C), and three previously described rare mutations, 1525-1(G>A), R560S, and 1898+1(G>T), were detected. Login to comment 10 ABCC7 p.Trp1282* Most of these are of low overall frequency, although certain ethnic groups or geographic areas exhibit elevated frequencies of particular mutations, the most notable of these being the W1282X mutation, which is present at a frequency of 60% in Ashkenazi Jewish CF patients (Shoshani et al., 1992). Login to comment 31 ABCC7 p.Tyr569Asp ABCC7 p.Gln98* RESULTS A total of 11 different mutations were detected in the Pakistani CF patients studied, including the five novel mutations: Y569D (Fig. 1), Q98X (Fig. 2), 296+12(T>C), 1161delC (Fig. 3), and 621+2 (T>C). Login to comment 38 ABCC7 p.Gln98* A novel polymorphism, 297-67(A>C) was detected in intron 2, on the same chromosome as the mutation Q98X. Login to comment 40 ABCC7 p.Arg560Ser Of the 14 patients, four were found to be homozygous for DeltaF508 and one was a compound heterozygote of DeltaF508 and the rare mutation R560S. Login to comment 42 ABCC7 p.Tyr569Asp ABCC7 p.Tyr569Asp ABCC7 p.Tyr569Asp Direct DNA cycle sequencing of CFTR exon 12 PCR products, showing the Y569D mutation. Lanes 14 (GATC) homozygote for the normal sequence (T at 1837), lanes 58 (GATC) heterozygote for Y569D (T and G at 1837), lanes 912 (GATC) homozygote for Y569D (G at 1837). Login to comment 46 ABCC7 p.Tyr569Asp DISCUSSION Novel Mutations Four unrelated patients were found to be homozygous for Y569D in exon 12. Login to comment 49 ABCC7 p.Gln98* Direct DNA cycle sequencing of CFTR exon 4 PCR products, showing the Q98X mutation. Login to comment 51 ABCC7 p.Gln98* Lanes 1,2,3,6,7,8,10,11,12 show the normal homozygote sequence (C at 424); lanes 4 and 9 shown the homozygote pattern for Q98X (T at 424) and lane 5 shows the heterozygote pattern (C and T at 424). Login to comment 58 ABCC7 p.Gln98* Three unrelated patients were found to be homozygous for Q98X. Login to comment 68 ABCC7 p.Arg560Ser Previously Described Rare Mutations Patient C453 was found to be heterozygous for the mutation R560S, which was originally detected in homozygous form in a Pakistani CF patient (Dr. Bruno Costes, pers. comm., 1995). Login to comment 77 ABCC7 p.Arg709* Patient CR93 was found to be heterozygous for the mutation R709X, which was originally detected in an Italian patient who is not of Pakistani origin (Elena Nicolis, pers. comm., 1996). Login to comment 82 ABCC7 p.Arg560Ser ABCC7 p.Tyr569Asp ABCC7 p.Tyr569Asp ABCC7 p.Tyr569Asp ABCC7 p.Tyr569Asp ABCC7 p.Tyr569Asp ABCC7 p.Tyr569Asp ABCC7 p.Tyr569Asp ABCC7 p.Tyr569Asp ABCC7 p.Gln98* ABCC7 p.Gln98* ABCC7 p.Gln98* ABCC7 p.Gln98* ABCC7 p.Gln98* ABCC7 p.Gln98* ABCC7 p.Arg709* region DeltaF508/DeltaF508 C61 North West DeltaF508/DeltaF508 C83 North West DeltaF508/DeltaF508 C218 North West DeltaF508/DeltaF508 C850 North West DeltaF508/R560S C453 North West DeltaF508/1898+1(G>T) CR726 Oxford Y569D/Y569D C289 North West Y569D/Y569D C707 North West Y569D/Y569D CR460 West Midlands Y569D/Y569D CR770 Glasgow Q98X/Q98X C813 North West Q98X/Q98X CR152 West Midlands Q98X/Q98X CR366 West Midlands 1525-1(G>A)/1525-1 CR97 West Midlands (G>A) 1525-1(G>A)/1525-1 CR780 Glasgow (G>A) 1525-1(G>A)/R709X CR93 West Midlands 296+12(T>C)/296+12 C11 North West (T>C) 296+12(T>C)/296+12 C886 North West (T>C) 1161delC/1161delC C633 North West 1161delC/1161delC CR382 Yorkshire 621+2(T>C)/621+2 C517 North West (T>C) 2184insA/2184insA CR153 West Midlands Unknown (?/?) Login to comment 87 ABCC7 p.Arg560Ser ABCC7 p.Tyr569Asp ABCC7 p.Tyr569Asp ABCC7 p.Gln98* ABCC7 p.Arg709* Current Methods of Analysis for CF Mutations Detected in Pakistani CF Patientsa Current Mutation Nucleotide change Exon or Intron Reference analysis methods 296+12(T>C) T>C at 296+12 intron 2 This study RE (Rsa I) Q98X C>T at 424 exon 4 This study SSCP/HD, SEQ 621+2(T>C) T>C at 621+2 intron 4 This study SSCP/HD, SEQ 1161delC deletion C at 1161 exon 7 This study PAGE, SEQ 1525-1(G>A) G>A at 1525-1 intron 9 Dörk et al. (1993) SSCP/HD, SEQ DeltaF508 deletion CTT at 1652 exon 10 Kerem et al. (1989) PAGE, CF(4)m R560S A>C at 1812 exon 12 Costes (p.c., 1993) SSCP/HD, DGGE, SEQ Y569D T>G at 1837 exon 12 This study SSCP/HD, DGGE, SEQ 1898+1(G>T) G>T at 1898+1 intron 12 Crawford et al. (1995) SSCP, SEQ 2184insA insertion A at 2184 exon 13 Dörk et al. (1994) SSCP/HD, SEQ R709X C>T at 2257 exon 13 Bonizzato et al. (1995) SSCP/HD, SEQ a Key: CF(4)m - CF(4)m PCR kit (Johnson and Johnson Ortho Clinical Diagnostics, Amersham, UK) SSCP/HD - Combined Single Stranded Conformational Polymorphism/Heteroduplex analysis; SEQ - direct DNA cycle sequencing; p.c. - pers. comm. consanguinityobservedinthispopulation.PatientsC61, C83, C218 (all homozygous for DeltaF508); CR382 (homozygous for 1161delC); C707 (homozygous for Y569D), C11 (homozygous for 296+12(T>C)) and CR694 (unknown genotype) are known to have consanguineous (first cousin) parents. Login to comment 93 ABCC7 p.Arg709* Of the five previously described rare mutations observed in this study, 2184insA and R709X have been detected in Pakistani CF patients as well as those who are native to the United Kingdom (Schwarz et al., 1995). Login to comment