The subcellular localization of SF2/ASF is regulated by direct interaction with SR protein kinases (SRPKs)

Koizumi, J., Okamoto, Y., Onogi, H., Mayeda, A., Krainer, A. R., Hagiwara, M. (April 1999) The subcellular localization of SF2/ASF is regulated by direct interaction with SR protein kinases (SRPKs). Journal of Biological Chemistry, 274 (16). pp. 11125-11131. ISSN 0021-9258

URL: http://www.ncbi.nlm.nih.gov/pubmed/10196197
DOI: 10.1074/jbc.274.16.11125

Abstract

Serine/arginine-rich (SR) proteins play an important role in constitutive and alternative pre-mRNA splicing. The C-terminal arginine serine domain of these proteins, such as SF2/ASF, mediates protein-protein interactions and is phosphorylated in vivo. Using glutathione S-transferase (GST)-SF2/ASF-affinity chromatography, the SF2/ASF kinase activity was co-purified from HeLa cells with a 95-kDa protein, which was recognized by an anti SR protein kinase (SRPK) 1 monoclonal antibody. Recombinant SRPK1 and SRPK2 bound to and phosphorylated GST-SF2/ASF in vitro. Phosphopeptide mapping showed that identical sites were phosphorylated in the pull-down kinase reaction with HeLa extracts and by recombinant SRPKs, Epitope-tagged SF2/ASF transiently expressed in COS7 cells co-immunoprecipitated with SRPKs, Deletion analysis mapped the phosphorylation sites to a region containing an (Arg-Ser)(8) repeat beginning at residue 204, and far-Western analysis showed that the region is required for binding of SRPKs to SF2/ASF, Further binding studies showed that SRPKs bound unphosphorylated SF2/ASF but did not bind phosphorylated SF2/ASF, Expression of an SRPK2 kinase-inactive mutant caused accumulation of SF2/ASF in the cytoplasm. These results suggest that the formation of complexes between SF2/ASF and SRPKs, which is influenced by the phosphorylation state of SF2/ASF, may have regulatory roles in the assembly and localization of this splicing factor.

Item Type: Paper
Uncontrolled Keywords: splice-site selection alternative 5' serine-rich cell-cycle rna phosphorylation domains purification factor-sf2 invitro
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > kinase
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein expression > phosphorylation
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > splice site
CSHL Authors:
Communities: CSHL labs > Krainer lab
Depositing User: Matt Covey
Date: April 1999
Date Deposited: 11 Dec 2013 19:22
Last Modified: 11 Dec 2013 19:22
Related URLs:
URI: http://repository.cshl.edu/id/eprint/28926

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