SIPP1, a novel pre-mRNA splicing factor and interactor of protein phosphatase-1

Biochem J. 2004 Feb 15;378(Pt 1):229-38. doi: 10.1042/BJ20030950.

Abstract

We have identified a polypeptide that was already known to interact with polyglutamine-tract-binding protein (PQBP)-1/Npw38 as a novel splicing factor and interactor of protein phosphatase-1, hence the name SIPP1 for splicing factor that interacts with PQBP-1 and PP1 (protein phosphotase 1). SIPP1 was inhibitory to PP1, and its inhibitory potency was increased by phosphorylation with protein kinase CK1. Two-hybrid and co-sedimentation analysis revealed that SIPP1 has two distinct PP1-binding domains and that the binding of SIPP1 with PP1 involves a RVXF (Arg-Val-Xaa-Phe) motif, which functions as a PP1-binding sequence in most interactors of PP1. Enhanced-green-fluorescent-protein-tagged SIPP1 was targeted exclusively to the nucleus and was enriched in the nuclear speckles, which represent storage/assembly sites of splicing factors. We have mapped a nuclear localization signal in the N-terminus of SIPP1, while the proline-rich C-terminal domain appeared to be required for its subnuclear targeting to the speckles. Finally, we found that SIPP1 is also a component of the spliceosomes and that a SIPP1-fragment inhibits splicing catalysis by nuclear extracts independent of its ability to interact with PP1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • COS Cells
  • Cell Line
  • Chlorocebus aethiops
  • DNA-Binding Proteins
  • HeLa Cells
  • Humans
  • Mice
  • Molecular Sequence Data
  • Phosphoprotein Phosphatases / chemistry
  • Phosphoprotein Phosphatases / metabolism*
  • Protein Phosphatase 1
  • Proteins / chemistry
  • Proteins / metabolism
  • Proteins / physiology*
  • RNA Precursors / metabolism*
  • RNA Splicing Factors
  • RNA Splicing*
  • RNA, Messenger / metabolism*
  • Spliceosomes / chemistry
  • Two-Hybrid System Techniques

Substances

  • DNA-Binding Proteins
  • Proteins
  • RNA Precursors
  • RNA Splicing Factors
  • RNA, Messenger
  • Wbp11 protein, mouse
  • Phosphoprotein Phosphatases
  • Protein Phosphatase 1