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Structural and functional analysis of essential pre-mRNA splicing factor Prp19p.

U-box-containing Prp19p is an integral component of the Prp19p-associated complex (the nineteen complex, or NTC) that is essential for activation of the spliceosome. Prp19p makes numerous protein-protein contacts with other NTC components and is required for NTC stability. Here we show that Prp19p forms a tetramer in vitro and in vivo and we map the domain required for its oligomerization to a central tetrameric coiled-coil. Biochemical and in vivo analyses are consistent with Prp19p tetramerization providing an interaction surface for a single copy of its binding partner, Cef1p. Electron microscopy showed that the isolated Prp19p tetramer is an elongated particle consisting of four globular WD40 domains held together by a central stalk consisting of four N-terminal U-boxes and four coiled-coils. These structural and functional data provide a basis for understanding the role of Prp19p as a key architectural component of the NTC.

Pubmed ID: 15601865 RIS Download

Mesh terms: Cell Cycle Proteins | Chromatography, Gel | Circular Dichroism | DNA Mutational Analysis | Dimerization | Genotype | Image Processing, Computer-Assisted | Immunoblotting | Immunoprecipitation | Microscopy, Electron | Models, Biological | Polymerase Chain Reaction | Protein Binding | Protein Conformation | Protein Structure, Tertiary | RNA Splicing | RNA Splicing Factors | RNA, Messenger | RNA-Binding Proteins | Recombinant Fusion Proteins | Saccharomyces cerevisiae | Saccharomyces cerevisiae Proteins | Schizosaccharomyces | Schizosaccharomyces pombe Proteins | Spliceosomes | Structure-Activity Relationship | Temperature | Two-Hybrid System Techniques | Ultracentrifugation

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Associated grants

  • Agency: NIGMS NIH HHS, Id: T32 GM008320
  • Agency: NIGMS NIH HHS, Id: P01 GM062580
  • Agency: NCI NIH HHS, Id: T32 CA009385
  • Agency: NCI NIH HHS, Id: T32 CA 09385
  • Agency: NIGMS NIH HHS, Id: R01 GM062112
  • Agency: NIGMS NIH HHS, Id: T32 GM 08320
  • Agency: NIGMS NIH HHS, Id: R01 GM 62112

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