title: Dystrophin nonsense mutations can generate alternative rescue transcripts in lymphocytes.
abstract: Secondary alterations in splicing have been reported to produce semi-functional mRNA from several nonsense mutations in the dystrophin gene. Disruptions of exonic splicing enhancers by single nucleotide changes are thought to underlie such alterations. The precise frequencies of such nonsense mutation-dependent splicing alterations, however, remain unknown. Here we analyzed the splicing patterns of dystrophin mRNA in lymphocytes from 38 patients with dystrophinopathies due to nonsense mutations in the dystrophin gene. In seven of the cases (18%), we observed partial skipping of the nonsense-encoding exon. Two of the seven cases, however, exhibited complex activation of a nonsense mutation-created splice site, which resulted in the generation of novel transcripts. Examination of cis-regulatory splicing elements through calculation of splicing probability scores and identification of potential splicing enhancer or silencer sequences failed to disclose a single cause for exon skipping. Remarkably, individual differences in splicing patterns were observed for cells from patients with identical nonsense mutations (C.5899C>T). Although five cases produced semi-functional dystrophin mRNAs, only one of these exhibited a mild clinical course. These results provide important insights about targets for exon skipping induced by candidate antisense oligonucleotides and for ribosomal read-through of nonsense mutations.
authors.affiliation: Department of Pediatrics, Kobe University Graduate School of Medicine, Kobe, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Nishiyama
authors.forename: Y
authors.initials: Y
authors.lastname: Takeshima
authors.forename: Z
authors.initials: Z
authors.lastname: Zhang
authors.forename: Y
authors.initials: Y
authors.lastname: Habara
authors.forename: T H T
authors.initials: TH
authors.lastname: Tran
authors.forename: M
authors.initials: M
authors.lastname: Yagi
authors.forename: M
authors.initials: M
authors.lastname: Matsuo
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D002675:Child, Preschool; D018389:Codon, Nonsense; D004252:DNA Mutational Analysis; D016189:Dystrophin / Q000235:genetics*; D006801:Humans; D007564:Japan; D008214:Lymphocytes / Q000378:metabolism*; D009136:Muscular Dystrophies / Q000235:genetics*; D012326:RNA Splicing
chemical_list: D018389:Codon, Nonsense; D016189:Dystrophin