title: Mutations in PRPF31 inhibit pre-mRNA splicing of rhodopsin gene and cause apoptosis of retinal cells.
abstract: Mutations in human PRPF31 gene have been identified in patients with autosomal dominant retinitis pigmentosa (adRP). To begin to understand mechanisms by which defects in this general splicing factor cause retinal degeneration, we examined the relationship between PRPF31 and pre-mRNA splicing of photoreceptor-specific genes. We used a specific anti-PRPF31 antibody to immunoprecipitate splicing complexes from retinal cells and identified the transcript of rhodopsin gene (RHO) among RNA species associated with PRPF31-containing complexes. Mutant PRPF31 proteins significantly inhibited pre-mRNA splicing of intron 3 in RHO gene. In primary retinal cell cultures, expression of the mutant PRPF31 proteins reduced rhodopsin expression and caused apoptosis of rhodopsin-positive retinal cells. This primary retinal culture assay provides an in vitro model to study photoreceptor cell death caused by PRPF31 mutations. Our results demonstrate that mutations in PRPF31 gene affect RHO pre-mRNA splicing and reveal a link between PRPF31 and RHO, two major adRP genes.
authors.affiliation: Department of Pediatrics, Cell and Developmental Biology, and Pharmacology, John F. Kennedy Center for Research on Human Development, Vanderbilt University Medical Center, Nashville, Tennessee 37232-8548, USA.
authors.forename: Liya
authors.initials: L
authors.lastname: Yuan
authors.forename: Mariko
authors.initials: M
authors.lastname: Kawada
authors.forename: Necat
authors.initials: N
authors.lastname: Havlioglu
authors.forename: Hao
authors.initials: H
authors.lastname: Tang
authors.forename: Jane Y
authors.initials: JY
authors.lastname: Wu
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000235:genetics / Q000502:physiology*; D061945:Caspase Inhibitors; D020169:Caspases / Q000502:physiology; D002478:Cells, Cultured; D005136:Eye Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D005786:Gene Expression Regulation; D006801:Humans; D051379:Mice; D009154:Mutation; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D012326:RNA Splicing / Q000235:genetics / Q000502:physiology*; D012160:Retina / Q000166:cytology* / Q000378:metabolism; D012174:Retinitis Pigmentosa / Q000235:genetics / Q000378:metabolism; D012243:Rhodopsin / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D061945:Caspase Inhibitors; D005136:Eye Proteins; C454222:PRPF31 protein, human; C103192:butyloxycarbonyl-O-methyl-aspartyl-fluoromethyl ketone; D012243:Rhodopsin; D020169:Caspases