The CFTR frameshift mutation 3905insT and its effect at transcript and protein level.
Cystic fibrosis (CF) is one of the most common genetic diseases in the Caucasian population and is characterized by chronic obstructive pulmonary disease, exocrine pancreatic insufficiency, and elevation of sodium and chloride concentrations in the sweat and infertility in men. The disease is caused by mutations in the CF transmembrane conductance regulator (CFTR) gene, which encodes a protein that functions as chloride channel at the apical membrane of different epithelia. Owing to the high genotypic and phenotypic disease heterogeneity, effects and consequences of the majority of the CFTR mutations have not yet been studied. Recently, the frameshift mutation 3905insT was identified as the second most frequent mutation in the Swiss population and found to be associated with a severe phenotype. The frameshift mutation produces a premature termination codon (PTC) in exon 20, and transcripts bearing this PTC are potential targets for degradation through nonsense-mediated mRNA decay (NMD) and/or for exon skipping through nonsense-associated alternative splicing (NAS). Using RT-PCR analysis in lymphocytes and different tissue types from patients carrying the mutation, we showed that the PTC introduced by the mutation does neither elicit a degradation of the mRNA through NMD nor an alternative splicing through NAS. Moreover, immunocytochemical analysis in nasal epithelial cells revealed a significantly reduced amount of CFTR at the apical membrane providing a possible molecular explanation for the more severe phenotype observed in F508del/3905insT compound heterozygotes compared with F508del homozygotes. However, further experiments are needed to elucidate the fate of the 3905insT CFTR in the cell after its biosynthesis.
Sanz
Javier
J
Department of Paediatrics, Division of Human Genetics, University of Bern, Bern, Switzerland.
von Känel
Thomas
T
Schneider
Mircea
M
Steiner
Bernhard
B
Schaller
André
A
Gallati
Sabina
S
D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000150:complications / Q000453:epidemiology / Q000235:genetics* / Q000473:pathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D016368:Frameshift Mutation; D006801:Humans; D008214:Lymphocytes / Q000473:pathology; D009154:Mutation; D014176:Protein Biosynthesis; D029424:Pulmonary Disease, Chronic Obstructive / Q000209:etiology / Q000235:genetics; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017384:Sequence Deletion; D013557:Switzerland / Q000453:epidemiology; D014158:Transcription, Genetic; D044465:White People / Q000706:statistics & numerical data
C505032:CFTR protein, human; D018076:DNA, Complementary; D012333:RNA, Messenger; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D012313:RNA