ABCMdb

PMID: 1283148

Lindner M, Wolf A, Moh B, Steinbach P, Kleihauer E, Bartram CR, Kulozik AE
The spectrum of CFTR mutations in south-west German cystic fibrosis patients.
Hum Genet. 1992 Nov;90(3):267-9., [PubMed]

Sentences

No. Mutations Sentence Comment

4 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Ala455Glu ABCC7 p.Arg553* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Arg1162* Another 5 mutations accounted for a further 12.5% (4% G542X; 3% R553X; 3% N1303K; 2% 1717-1 G--~A; 0.5% G551D) whereas 6 mutations (R117H, A455E, AI507, $549I, $549N, and R1162X) were not found. Login to comment 7 ABCC7 p.Asn1303Lys ABCC7 p.Gly542* Two patients, age 6 and 25 years, were compound heterozygotes for G542X and N1303K. Login to comment 8 ABCC7 p.Asn1303Lys The clinical features of the 6 year old were characterised by severe gastrointestinal and as yet only mild pulmonary complications whereas the 25 year old manifested severe pulmonary and gastrointestinal symptoms indicating that the N1303K mutation of the C-terminal CFTR nucleotide binding fold significantlyimpairs protein function in both the pancreas and the lungs. Login to comment 25 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Asn1303Lys ABCC7 p.Gly542* Sequences derived from Dean et al. (1990); Kerem et al. (1990); Riordan et al. (1989); Guillermit et al. (1990); Cutting et al. (1990) Exon PCR primers Mutation Allele specific probes 4 5'-AGTCACCAAAGCAGTACAGC-3' R117H N: 5'-GCTATTCTCATCTGCATTCC-3' M: 9 5'-TAATGGATCATGGGCCATGT-3' A455 E N: 5'-ACAGTG~ITGAATGTGGTGCA-3' M: 5'-GTTTTCCTGGATTATGCCTGGCAC-3' N: 5'-GTTGGCATGCTTTGATGACGCTTC-3' M: t0 11 5'-CAACTGTGGTTAAAGCAATAGTGT-3' 5'-GCACAGATTCTGAGTAACCATAAT-3' 5'-GAGGAACGCTCTATC-3' 5'-GAGGAACACTCTATC-3' 5'-GTTGTTGGCG GTI'GCT-3' 5'-GTTGTTGGAG GTTGCT-3' zXF508 5'-CACCAAAGATGATATTTTC-3' 5'-AACACCAATGATATTTTCTT-3' AI507 1717-1G--+A N: 5'-TTGGTAATAGGACATCTCCA-3' M: 5'-TTGGTAATAAGACATCTCCA-3' G542X N: 5'-CCACCTTCTCCAAGAACTAT-3' M: 5'-CCACCTTCTCAAAGAACTAT-3 G551D N: 5'-CTCGTTGACCTCCACTCAGT-3' M: 5'-CTCGTTGATCTCCACTCAGT-3' 19 5'-GCCCGACAAATAACCAAGTGA-3' R 1162X 5'-GCTAACACATTGCTFCAGGCT-3' 21 5'-AATGTTCACAAGGGACTCCA-3' N1303K 5'-CAAAAGTACCTGTTGCTCCA-3' N: 5'-AGGGATCCAAGTTTTTTCTA-3 ' M: 5'-AGGGATCCAACTTTTTTCTA-3' Materials and methods Determination of the CFTR genotype All mutation analyses were performed on Taq-PCR (polymerase chain reaction) amplified DNA fragments (30 cycles at 92~176 72~ for 30/60/90 s) using primers as shown in Table 1 (Sambrook et al. 1989). Login to comment 26 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Ala455Glu ABCC7 p.Gly542* The R117H, A455E, 1717-1G~A, AF508, G542X, G551D, and NI303K mutations were identified by allele specific oligonucleotide hybridisation (ASO, Sambrook et al. 1989) using 32p-labelled probes specific for the normal or the mutated sequence (Table 1). Login to comment 27 ABCC7 p.Arg553* The Rl162X, $549N, $549I and the R553X mutations were identified by DdeI or HincII restriction analysis (Gasparini et al. 1991: Cutting et al. 1990). Login to comment 29 ABCC7 p.Gly551Asp In addition to ASO, the G551D, AF508 and AI507 mutations were identified by HincII and Mbol restriction analysis (Cutting et al. 1990) or by polyacrylamide gel electrophoresis, respectively (Scheffer et al. 1989). Login to comment 33 ABCC7 p.Arg553* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* The other five mutations were found on a further 12% of the CF chromosomes, the most numerous of these being G542X, R553X, and N1303K (Table 2).This distribution is similar to that found in a group of patients from the north of Germany, although in contrast to our Table 2. Login to comment 34 ABCC7 p.Gly551Asp ABCC7 p.Ala455Glu ABCC7 p.Arg553* ABCC7 p.Arg553* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Gly542* ABCC7 p.Arg1162* Mutations observed in 220 defective cystic fibrosis transmembrane conductance regulator (CFTR) alleles Mutation n (%) Rll7H 0 A455E 0 AI507 0 AF508 147 (67) 1717-1G--*A 4 (2) G542X 9 (4) $549I 0 $549N 0 G551D 1 (<0.5) R553X 7 (3) R1162X 0 N1303K 6 (3) Unknown 46 (21) Total 220 (100) patients in that group R553X was considerably more common than G542X (Plieth et al. 1992). Login to comment 36 ABCC7 p.Asn1303Lys ABCC7 p.Gly542* Fifty-four patients (49%) were homozygous for A508, 18 (16.5%) were compound heterozygotes for AF508 and one of the rarer mutations, and 2 were compound heterozygotes for G542X and N1303K. Login to comment 40 ABCC7 p.Gly551Asp ABCC7 p.Arg553* ABCC7 p.Asn1303Lys ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Gly542* ABCC7 p.Gly542* Genotype of 110 CF patients Genotype n (%) F508/F508a 54 (49) F508/1717-1G---~A 4 (3.5) F508/G542X 3 (3) F508/G551D 1 (1) F508/R553Xb 5 (4.5) F508/N1303K 4 (3.5) F508/unknown c 22 (20) G542X/N1303K 2 (2) G542X/unknown 4 (3.5) R553X/unknown 2 (2) Unknown/unknown d 9 (8) Total 110 (100) a Includes one Italian patient b Includes one patient with a French father and a German mother c Includes two Italian and one French patient Includes one Greek and one Turkish patient sorption is also indicated by repeated low stool chymotrypsin levels (<0.3 U/g) and a reduced excretion (< 10%) of para-amino benzoic acid (PABA) following a standardised oral challenge. Login to comment 45 ABCC7 p.Asn1303Lys ABCC7 p.Asn1303Lys ABCC7 p.Arg334* Although the small number of patients with this genotype does not allow any general statements about the phenotypic expression of this particular genotype or the effect of the N1303K mutation on CFTR function it is interesting to note that another compound heterozygous patient for the N1303K mutation and another nonsense mutation (R334X) has also been reported to be characterised by gastrointestinal and pulmonary involvement (Gasparini et al. 1991). Login to comment 48 ABCC7 p.Arg117His ABCC7 p.Ala455Glu We are grateful to Professor Kubanek for his continuous support, to Professor Kaiser, Pforzheim for sending blood samples and providing clinical information on some of his patients, and to Dr.M. Schwarz, Royal Manchester Children's Hospital, for providing allele specific oligonucleotides for the R117H, A455E and 1717-1G--~A mutations together with the appropriate controls. Login to comment