Novel SR-rich-related protein clasp specifically interacts with inactivated Clk4 and induces the exon EB inclusion of Clk

Rieko Katsu; Hiroshi Onogi; Kazuhiro Wada; Yasushi Kawaguchi; Masatoshi Hagiwara

doi:10.1074/jbc.M206504200

Novel SR-rich-related protein clasp specifically interacts with inactivated Clk4 and induces the exon EB inclusion of Clk

J Biol Chem. 2002 Nov 15;277(46):44220-8. doi: 10.1074/jbc.M206504200. Epub 2002 Aug 6.

Authors

Rieko Katsu¹, Hiroshi Onogi, Kazuhiro Wada, Yasushi Kawaguchi, Masatoshi Hagiwara

Affiliation

¹ Department of Functional Genomics, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Japan.

PMID: 12169693
DOI: 10.1074/jbc.M206504200

Abstract

We identified a novel serine/arginine (SR)-rich-related protein as a binding partner of Clk4 mutant lacking kinase activity (Clk4 K189R) in the two-hybrid screen and designated it Clasp (Clk4-associating SR-related protein). Northern blot analysis revealed that Clasp mRNA was highly expressed in brain, and in situ hybridization of a mouse brain sagittal section hybridized with antisense probes revealed that both Clasp and Clk4 mRNAs were expressed in the hippocampus, the cerebellum, and the olfactory bulb. Two forms of Clasp were produced by a frameshift following alternative splicing. The staining of an HA-tagged short form of Clasp (ClaspS) showed a nucleoplasmic pattern, while the long form of Clasp (ClaspL) was localized as nuclear dots. In vitro protein interaction assay demonstrated that Clk4 K189R was bound to Clasp while wild Clk4 was not. Overexpression of ClaspL promoted accumulation of Clk4 K189R in the nuclear dots and the exon EB inclusion from CR-1 and CR-2 pre-mRNA of Clk1. These data suggest that Clasp is a binding partner of Clk4 and may be involved in the regulation of the activity of Clk kinase family.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alternative Splicing
Amino Acid Sequence
Animals
Arginine / chemistry
Base Sequence
Blotting, Northern
COS Cells
Cell Nucleus / metabolism
Cerebellum / metabolism
Cloning, Molecular
Exons
Frameshift Mutation
Glutathione Transferase / metabolism
Hippocampus / metabolism
In Situ Hybridization
Mice
Microscopy, Fluorescence
Molecular Sequence Data
Nuclear Proteins / biosynthesis*
Nuclear Proteins / chemistry*
Olfactory Bulb / metabolism
Phosphorylation
Plasmids / metabolism
Protein Binding
Protein Serine-Threonine Kinases / genetics*
Protein Serine-Threonine Kinases / metabolism
Protein-Tyrosine Kinases / genetics*
Protein-Tyrosine Kinases / metabolism
RNA, Messenger / metabolism
Recombinant Proteins / metabolism
Reverse Transcriptase Polymerase Chain Reaction
Serine / chemistry
Tissue Distribution
Two-Hybrid System Techniques

Substances

Clasp protein, mouse
Nuclear Proteins
RNA, Messenger
Recombinant Proteins
Serine
Arginine
Glutathione Transferase
Clk dual-specificity kinases
Protein-Tyrosine Kinases
Protein Serine-Threonine Kinases

Associated data

GENBANK/AB080582
GENBANK/AB080583