Regulated tissue-specific expression of antagonistic pre-mRNA splicing factors

Hanamura, A., Caceres, J. F., Mayeda, A., Franza, B. R., Krainer, A. R. (April 1998) Regulated tissue-specific expression of antagonistic pre-mRNA splicing factors. RNA, 4 (4). pp. 430-444. ISSN 1355-8382

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URL: http://www.ncbi.nlm.nih.gov/pubmed/9630249

Abstract

The SR proteins are essential metazoan pre-mRNA splicing factors that can also influence the selection of alternative 5' splice sites in a concentration-dependent manner. Their activity in alternative splicing in vitro is antagonized by members of the hnRNP A/B family of proteins. The opposite effects of members of these two families of antagonistic splicing factors in vitro and upon overexpression in vivo suggest that changes in their relative levels may be a natural mechanism for the regulation of alternative splicing in vivo. One prediction of this model is that the ratios of these antagonists should vary in different cell types and in other situations in which cellular or viral transcripts are differentially spliced. We raised monoclonal antibodies specific for SFS/ASF and used them to measure the abundance of SFS/ASF protein and its isoforms, its phosphorylation state in vivo and during splicing in vitro, and its association with the spliceosome. SF2/ASF exists predominantly or exclusively in a highly phosphorylated state in vivo in all cell types examined, and unphosphorylated protein was not detectable. Unphosphorylated recombinant SFS/ASF becomes rapidly phosphorylated under splicing conditions in HeLa cell extracts and associates stably with one or more exons of beta-globin pre-mRNA. This interaction appears to persist through the splicing reaction and SF2/ASF remains bound to spliced mRNA. We compared the distribution of SFS/ASF to that of its antagonist, hnRNP Al, in different rat tissues and in immortal and transformed cell lines. We found that the protein levels of these antagonistic splicing factors vary naturally over a very wide range, supporting the notion that changes in the ratio of these proteins can affect alternative splicing of a variety of pre-mRNAs in vivo.

Item Type: Paper
Uncontrolled Keywords: hnRNP A1 monoclonal antibodies phosphorylation pre-mRNA splicing factors SF2/ASF SR proteins rna-binding proteins transformed human-cells messenger-rna premessenger-rna sr proteins in-vivo site selection hnrnp protein drosophila development monoclonal-antibody
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 > DNA, RNA structure, function, modification > pre-mRNA
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNA splicing
CSHL Authors:
Communities: CSHL labs > Krainer lab
Depositing User: Matt Covey
Date: April 1998
Date Deposited: 11 Dec 2013 15:37
Last Modified: 11 Dec 2013 15:37
PMCID: PMC1369629
Related URLs:
URI: http://repository.cshl.edu/id/eprint/28935

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