PUBLICATION

Identification of a PRPF4 Loss-of-Function Variant That Abrogates U4/U6.U5 Tri-snRNP Integration and Is Associated with Retinitis Pigmentosa

Authors
Linder, B., Hirmer, A., Gal, A., Rüther, K., Bolz, H.J., Winkler, C., Laggerbauer, B., Fischer, U.
ID
ZDB-PUB-141111-2
Date
2014
Source
PLoS One   9: e111754 (Journal)
Registered Authors
Winkler, Christoph
Keywords
none
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Western
  • Embryo, Nonmammalian/metabolism
  • Gangliosides/metabolism
  • Gene Components
  • HEK293 Cells
  • Humans
  • Molecular Sequence Data
  • Mutation, Missense/genetics*
  • Nuclear Proteins/metabolism
  • Pedigree
  • Retinitis Pigmentosa/genetics*
  • Ribonucleoprotein, U4-U6 Small Nuclear/genetics*
  • Ribonucleoprotein, U4-U6 Small Nuclear/metabolism
  • Sequence Analysis, DNA
  • Spliceosomes/genetics*
  • Spliceosomes/metabolism
  • Zebrafish
PubMed
25383878 Full text @ PLoS One
Abstract
Pre-mRNA splicing by the spliceosome is an essential step in the maturation of nearly all human mRNAs. Mutations in six spliceosomal proteins, PRPF3, PRPF4, PRPF6, PRPF8, PRPF31 and SNRNP200, cause retinitis pigmentosa (RP), a disease characterized by progressive photoreceptor degeneration. All splicing factors linked to RP are constituents of the U4/U6.U5 tri-snRNP subunit of the spliceosome, suggesting that the compromised function of this particle may lead to RP. Here, we report the identification of the p.R192H variant of the tri-snRNP factor PRPF4 in a patient with RP. The mutation affects a highly conserved arginine residue that is crucial for PRPF4 function. Introduction of a corresponding mutation into the zebrafish homolog of PRPF4 resulted in a complete loss of function in vivo. A series of biochemical experiments suggested that p.R192H disrupts the binding interface between PRPF4 and its interactor PRPF3. This interferes with the ability of PRPF4 to integrate into the tri-snRNP, as shown in a human cell line and in zebrafish embryos. These data suggest that the p.R192H variant of PRPF4 represents a functional null allele. The resulting haploinsufficiency of PRPF4 compromises the function of the tri-snRNP, reinforcing the notion that this spliceosomal particle is of crucial importance in the physiology of the retina.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping