title: Aberrant splicing in the PKD2 gene as a cause of polycystic kidney disease.
abstract: It is estimated that approximately 15% of families with autosomal dominant polycystic kidney disease (ADPKD) have mutations in PKD2. Identification of these mutations is central to identifying functionally important regions of gene and to understanding the mechanisms underlying the pathogenesis of the disorder. The current study describes mutations in six type 2 ADPKD families. Two single base substitution mutations discovered in the ORF in exon 14 constitute the most COOH-terminal pathogenic variants described to date. One of these mutations is a nonsense change and the other encodes an apparent missense variant. Reverse transcription-PCR from patient lymphoblast RNA showed that, in addition, both mutations resulted in out-of-frame splice variants by activating cryptic splice sites via different mechanisms. The apparent missense variant produced such a strong splicing signal that the processed transcript from the mutant chromosome did not contain any of the normally spliced, missense product. A third mutation, a nonconservative missense change effecting a negatively charged residue in the third transmembrane span, is likely pathogenic and defines a highly conserved residue consistent with a potential channel subunit function for polycystin-2. The remaining three mutations included two frame shifts resulting from deletion of one or two bases in exons 6 and 10, respectively, and a nonsense mutation due to a single base substitution in exon 4. The study also defined a novel intragenic polymorphism in exon 1 that will be useful in analyzing "second hits" in PKD2. Finally, the study demonstrates that there are reduced levels of normal polycystin-2 protein in lymphoblast lines from PKD2-affected individuals and that truncated mutant polycystin-2 cannot be detected in patient lymphoblasts, suggesting that the latter may be unstable in at least some tissues. The mutations described will serve as critical reagents for future functional studies in PKD2.
authors.affiliation: Department of Medicine, Albert Einstein College of Medicine, Bronx, New York, USA.
authors.forename: D M
authors.initials: DM
authors.lastname: Reynolds
authors.forename: T
authors.initials: T
authors.lastname: Hayashi
authors.forename: Y
authors.initials: Y
authors.lastname: Cai
authors.forename: B
authors.initials: B
authors.lastname: Veldhuisen
authors.forename: T J
authors.initials: TJ
authors.lastname: Watnick
authors.forename: X M
authors.initials: XM
authors.lastname: Lens
authors.forename: T
authors.initials: T
authors.lastname: Mochizuki
authors.forename: F
authors.initials: F
authors.lastname: Qian
authors.forename: Y
authors.initials: Y
authors.lastname: Maeda
authors.forename: L
authors.initials: L
authors.lastname: Li
authors.forename: R
authors.initials: R
authors.lastname: Fossdal
authors.forename: E
authors.initials: E
authors.lastname: Coto
authors.forename: G
authors.initials: G
authors.lastname: Wu
authors.forename: M H
authors.initials: MH
authors.lastname: Breuning
authors.forename: G G
authors.initials: GG
authors.lastname: Germino
authors.forename: D J
authors.initials: DJ
authors.lastname: Peters
authors.forename: S
authors.initials: S
authors.lastname: Somlo
mesh_terms: D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D009154:Mutation; D016891:Polycystic Kidney, Autosomal Dominant / Q000235:genetics*; D011110:Polymorphism, Genetic; D012326:RNA Splicing; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050396:TRPP Cation Channels
chemical_list: D008565:Membrane Proteins; D050396:TRPP Cation Channels