title: Detection of two different nonsense mutations in exon 44 of the PKD1 gene in two unrelated Italian families with severe autosomal dominant polycystic kidney disease.
abstract: Sixty-seven Italian patients with autosomal dominant polycystic kidney disease (ADPKD) were screened for mutations in the PKD1 gene. We used PCR, heteroduplex and single-strand conformation polymorphism DNA analysis, and automated DNA sequencing for exons 35, 36, 38, 44 and 45. We detected abnormal heteroduplexes in affected individuals from two unrelated families with clinically severe ADPKD phenotype. These changes were absent in other, unaffected members, as well as in the probands of the other families studied. DNA sequencing revealed in both cases different C to T transitions in exon 44, which created premature stop codons. Both mutations altered restriction sites, and the abnormal patterns were observed in all the affected family members. RT-PCR performed on lymphocyte mRNA showed that both the mutant and the normal transcript are represented. To our knowledge these are the first nonsense mutations described in the PKD1 gene.
authors.affiliation: Institute of Genetics, University of Verona School of Medicine, University Hospital Polyclinic Borgo Roma, Italy.
authors.forename: A E
authors.initials: AE
authors.lastname: Turco
authors.forename: S
authors.initials: S
authors.lastname: Rossetti
authors.forename: E
authors.initials: E
authors.lastname: Bresin
authors.forename: S
authors.initials: S
authors.lastname: Corrà
authors.forename: G
authors.initials: G
authors.lastname: Restagno
authors.forename: A
authors.initials: A
authors.lastname: Carbonara
authors.forename: O
authors.initials: O
authors.lastname: De Prisco
authors.forename: L
authors.initials: L
authors.lastname: Gammaro
authors.forename: G
authors.initials: G
authors.lastname: Maschio
authors.forename: P F
authors.initials: PF
authors.lastname: Pignatti
mesh_terms: D005091:Exons; D006801:Humans; D009154:Mutation; D009692:Nucleic Acid Heteroduplexes; D016891:Polycystic Kidney, Autosomal Dominant / Q000235:genetics*; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D011506:Proteins / Q000235:genetics*; D050396:TRPP Cation Channels; D014158:Transcription, Genetic
chemical_list: D009692:Nucleic Acid Heteroduplexes; D011506:Proteins; D050396:TRPP Cation Channels