ABCMdb

PMID: 10050655

Lissens W, Mahmoud KZ, El-Gindi E, Abdel-Sattar A, Seneca S, Van Steirteghem A, Liebaers I
Molecular analysis of the cystic fibrosis gene reveals a high frequency of the intron 8 splice variant 5T in Egyptian males with congenital bilateral absence of the vas deferens.
Mol Hum Reprod. 1999 Jan;5(1):10-3., [PubMed]

Sentences

No. Mutations Sentence Comment

4 ABCC7 p.Leu541Pro ABCC7 p.Leu541Pro CFTR mutations or the IVS-5T variant were found neither in the remaining four patients with associated renal abnormalities nor in the spouses of the 20 CBAVD patients. However, one patient was homozygous for a leucine to proline substitution at amino acid position 541 (L541P) of the CFTR. Login to comment 32 ABCC7 p.Gly551Asp ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* Initially, eight of the males` samples were analysed by a commercial kit allowing the detection of eight mutations in the CFTR gene: ∆F508, ∆I507, G542X, G551D, R553X, 1717-1G→A, W1282X and N1303K (INNO-LiPA CF, Innogenetics, Zwijnaarde, Belgium). Login to comment 33 ABCC7 p.Arg117His ABCC7 p.Arg334Trp ABCC7 p.Arg1162* Later on, all samples, including the previous ones, were studied by using the CF(12)m polymerase chain reaction (PCR) kit (Zeneca Diagnostics, Abingdon, UK) allowing the detection of the mutations present in the INNO-LiPA CF kit (with the exception of the ∆I507 mutation) and the R117H, R1162X, R334W, 621ϩ1G→T and 3849ϩ10 kbC→T mutations. Login to comment 35 ABCC7 p.Leu541Pro Screening for the presence of the L541P change was done by restriction enzyme digestion of the PCR amplified exon 11 (with primers 11i5 and 11i3) with ScrFI. Login to comment 41 ABCC7 p.Leu541Pro ABCC7 p.Leu541Pro Screening for the presence of the 1754T→C substitution (amino acid change leucine to proline at position 541 or L541P) in exon 11 of the CFTR gene by ScrFI restriction analysis. Login to comment 52 ABCC7 p.Gly542* None of the other individuals showed a mutation except the male with the ectopic pelvic kidney (patient ii in the previous paragraph) who had neither a normal nor a mutated band at the G542X site when studied by the INNO-LiPA CF kit. Login to comment 53 ABCC7 p.Gly551Asp ABCC7 p.Arg553* In this patient, normal bands were visible at the sites of the other mutations localized in exon 11 (1717-1G→A, G551D, R553X) thereby excluding a deletion of exon 11. Login to comment 54 ABCC7 p.Leu541Pro ABCC7 p.Leu541Pro PCR amplification and sequencing of the whole coding region of exon 11 showed that he was homozygous for a T to C substitution at cDNA position 1754 of the CFTR (1754T→C), predicting an amino acid change of leucine to proline at amino acid position 541 (L541P). Login to comment 55 ABCC7 p.Gly542* This base substitution probably inhibits the binding of the exon 11 PCR fragment to the normal (and mutant) sequence at the G542X locations in the INNO-LiPA CF kit. Login to comment 56 ABCC7 p.Leu541Pro Since this substitution would probably not be detected in heterozygotes with a normal and an L541P allele, another approach, based on the creation of an additional ScrFI restriction site in the PCR fragment defined by primers 11i5 and 11i3, was used to screen for it in the other individuals (Figure 1). Login to comment 63 ABCC7 p.Leu541Pro bIncluding the L541P homozygote and heterozygote female. Login to comment 65 ABCC7 p.Leu541Pro The L541P change was found on a 7T background. Login to comment 83 ABCC7 p.Leu541Pro These results are in agreement with a proposed different, but as yet unidentified, aetiology of the condition of CBAVD in these patients. However, one patient was homozygous for an L541P substitution in the CFTR gene, a change that so far has not been described either as a polymorphism or as a mutation. Login to comment 84 ABCC7 p.Leu541Pro The L541P substitution is in the first nucleotide binding fold of the CFTR and would be localized, according to the model of Hyde et al. (1990), in the third β-sheet of the ATP-binding cassette (ABC) of the protein. Login to comment