Abstract
Several ubiquitously expressed genes encoding pre-mRNA splicing factors have been associated with autosomal dominant retinitis pigmentosa (adRP), including PRPF31, PRPF3 and PRPF8. Molecular mechanisms by which defects in pre-mRNA splicing factors cause photoreceptor degeneration are not clear. To investigate the role of pre-mRNA splicing in photoreceptor gene expression and function, we have begun to search for photoreceptor genes whose pre-mRNA splicing is affected by mutations in PRPF31. Using an immunoprecipitation-coupled-microarray method, we identified a number of transcripts associated with PRPF31-containing complexes, including peripherin/RDS, FSCN2 and other photoreceptor-expressed genes. We constructed minigenes to study the effects of PRPF31 mutations on the pre-mRNA splicing of these photoreceptor specific genes. Our experiments demonstrated that mutant PRPF31 significantly inhibited pre-mRNA splicing of RDS and FSCN2. These observations suggest a functional link between ubiquitously expressed and retina-specifically expressed adRP genes. Our results indicate that PRPF31 mutations lead to defective pre-mRNA splicing of photoreceptor-specific genes and that the ubiquitously expressed adRP gene, PRPF31, is critical for pre-mRNA splicing of a subset of photoreceptor genes. Our results provide an explanation for the photoreceptor-specific phenotype of PRPF31 mutations.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Alternative Splicing / genetics
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Animals
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Animals, Newborn
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Carrier Proteins / biosynthesis
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Carrier Proteins / genetics
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Cell Line
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Cells, Cultured
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Chickens
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Chromosome Disorders / genetics
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Chromosome Disorders / metabolism
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Chromosome Disorders / physiopathology
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Eye Proteins / genetics*
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Gene Expression Regulation, Developmental / genetics
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Genes, Dominant / genetics
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Genetic Predisposition to Disease / genetics
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Humans
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Intermediate Filament Proteins / genetics
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Intermediate Filament Proteins / metabolism
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Membrane Glycoproteins / genetics
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Membrane Glycoproteins / metabolism
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Mice
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Mice, Inbred Strains
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Microfilament Proteins / biosynthesis
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Microfilament Proteins / genetics
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Mutation / genetics*
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Nerve Tissue Proteins / genetics
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Nerve Tissue Proteins / metabolism
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Oligonucleotide Array Sequence Analysis
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Peripherins
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Photoreceptor Cells / metabolism*
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Photoreceptor Cells / physiopathology
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RNA Precursors / genetics
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RNA Precursors / metabolism*
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RNA Splicing / genetics*
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Retinitis Pigmentosa / genetics*
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Retinitis Pigmentosa / metabolism
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Retinitis Pigmentosa / physiopathology
Substances
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Carrier Proteins
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Eye Proteins
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Intermediate Filament Proteins
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Membrane Glycoproteins
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Microfilament Proteins
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Nerve Tissue Proteins
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PRPF31 protein, human
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PRPH protein, human
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PRPH2 protein, human
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Peripherins
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Prph2 protein, mouse
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RNA Precursors
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fascin