title: Premature termination codons in PRPF31 cause retinitis pigmentosa via haploinsufficiency due to nonsense-mediated mRNA decay.
abstract: Dominant mutations in the gene encoding the mRNA splicing factor PRPF31 cause retinitis pigmentosa, a hereditary form of retinal degeneration. Most of these mutations are characterized by DNA changes that lead to premature termination codons. We investigated 6 different PRPF31 mutations, represented by single-base substitutions or microdeletions, in cell lines derived from 9 patients with dominant retinitis pigmentosa. Five of these mutations lead to premature termination codons, and 1 leads to the skipping of exon 2. Allele-specific measurement of PRPF31 transcripts revealed a strong reduction in the expression of mutant alleles. As a consequence, total PRPF31 protein abundance was decreased, and no truncated proteins were detected. Subnuclear localization of the full-length PRPF31 that was present remained unaffected. Blocking nonsense-mediated mRNA decay significantly restored the amount of mutant PRPF31 mRNA but did not restore the synthesis of mutant proteins, even in conjunction with inhibitors of protein degradation pathways. Our results indicate that most PRPF31 mutations ultimately result in null alleles through the activation of surveillance mechanisms that inactivate mutant mRNA and, possibly, proteins. Furthermore, these data provide compelling evidence that the pathogenic effect of PRPF31 mutations is likely due to haploinsufficiency rather than to gain of function.
authors.affiliation: Department of Medical Genetics, University of Lausanne, Lausanne, Switzerland.
authors.forename: Thomas
authors.initials: T
authors.lastname: Rio Frio
authors.forename: Nicholas M
authors.initials: NM
authors.lastname: Wade
authors.forename: Adriana
authors.initials: A
authors.lastname: Ransijn
authors.forename: Eliot L
authors.initials: EL
authors.lastname: Berson
authors.forename: Jacques S
authors.initials: JS
authors.lastname: Beckmann
authors.forename: Carlo
authors.initials: C
authors.lastname: Rivolta
mesh_terms: D000328:Adult; D000483:Alleles; D002460:Cell Line; D018389:Codon, Nonsense / Q000235:genetics*; D005136:Eye Proteins / Q000235:genetics* / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D006239:Haplotypes; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation / Q000235:genetics; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012174:Retinitis Pigmentosa / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D014158:Transcription, Genetic / Q000235:genetics
chemical_list: D018389:Codon, Nonsense; D005136:Eye Proteins; C454222:PRPF31 protein, human; D012333:RNA, Messenger