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PMID: 10022843 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The splicing factor-associated protein, p32, regulates RNA splicing by inhibiting ASF/SF2 RNA binding and phosphorylation.

The EMBO journal ·Vol. 18 ·No. 4 ·1999-02-15 ·Pages 1014-24

Petersen-Mahrt SK, Estmer C, Ohrmalm C, Matthews DA, Russell WC, Akusjärvi G

Abstract

The cellular protein p32 was isolated originally as a protein tightly associated with the essential splicing factor ASF/SF2 during its purification from HeLa cells. ASF/SF2 is a member of the SR family of splicing factors, which stimulate constitutive splicing and regulate alternative RNA splicing in a positive or negative fashion, depending on where on the pre-mRNA they bind. Here we present evidence that p32 interacts with ASF/SF2 and SRp30c, another member of the SR protein family. We further show that p32 inhibits ASF/SF2 function as both a splicing enhancer and splicing repressor protein by preventing stable ASF/SF2 interaction with RNA, but p32 does not block SRp30c function. ASF/SF2 is highly phosphorylated in vivo, a modification required for stable RNA binding and protein-protein interaction during spliceosome formation, and this phosphorylation, either through HeLa nuclear extracts or through specific SR protein kinases, is inhibited by p32. Our results suggest that p32 functions as an ASF/SF2 inhibitory factor, regulating ASF/SF2 RNA binding and phosphorylation. These findings place p32 into a new group of proteins that control RNA splicing by sequestering an essential RNA splicing factor into an inhibitory complex.

MeSH Terms
Carrier Proteins Cytoplasm/genetics Gene Expression Regulation/genetics Globins/genetics HeLa Cells Humans Hyaluronan Receptors Mitochondrial Proteins Nuclear Proteins/genetics Phosphoproteins/genetics Phosphorylation Proteins/genetics RNA Splicing/genetics RNA-Binding Proteins/genetics Recombinant Proteins/genetics Serine-Arginine Splicing Factors Transfection/genetics
Chemicals
C1QBP protein, human Carrier Proteins Hyaluronan Receptors Mitochondrial Proteins Nuclear Proteins Phosphoproteins Proteins RNA-Binding Proteins Recombinant Proteins Serine-Arginine Splicing Factors Globins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Petersen-Mahrt S K
Department of Medical Biochemistry and Microbiology, BMC, Uppsala University, S-751 23 Uppsala, Sweden.
Estmer C
Ohrmalm C
Matthews D A
Russell W C
Akusjärvi G
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/18.4.1014
Published
1999-02-15
Pages
1014-24
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1171193
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/18.4.1014 PMC Full Text View journal details
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