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PMID: 9671462 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cyclin E associates with components of the pre-mRNA splicing machinery in mammalian cells.

Molecular and cellular biology ·Vol. 18 ·No. 8 ·1998-08-00 ·Pages 4526-36

Seghezzi W, Chua K, Shanahan F, Gozani O, Reed R, Lees E

Abstract

Cyclin E-cdk2 is a critical regulator of cell cycle progression from G1 into S phase in mammalian cells. Despite this important function little is known about the downstream targets of this cyclin-kinase complex. Here we have identified components of the pre-mRNA processing machinery as potential targets of cyclin E-cdk2. Cyclin E-specific antibodies coprecipitated a number of cyclin E-associated proteins from cell lysates, among which are the spliceosome-associated proteins, SAP 114, SAP 145, and SAP 155, as well as the snRNP core proteins B' and B. The three SAPs are all subunits of the essential splicing factor SF3, a component of U2 snRNP. Cyclin E antibodies also specifically immunoprecipitated U2 snRNA and the spliceosome from splicing extracts. We demonstrate that SAP 155 serves as a substrate for cyclin E-cdk2 in vitro and that its phosphorylation in the cyclin E complex can be inhibited by the cdk-specific inhibitor p21. SAP 155 contains numerous cdk consensus phosphorylation sites in its N terminus and is phosphorylated prior to catalytic step II of the splicing pathway, suggesting a potential role for cdk regulation. These findings provide evidence that pre-mRNA splicing may be linked to the cell cycle machinery in mammalian cells.

MeSH Terms
Amino Acid Sequence CDC2-CDC28 Kinases Cyclin E/metabolism Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinases/metabolism Humans Molecular Sequence Data Phosphoproteins/metabolism Precipitin Tests Protein Serine-Threonine Kinases/metabolism RNA Precursors RNA Splicing RNA Splicing Factors Ribonucleoprotein, U2 Small Nuclear/metabolism Spliceosomes/metabolism Substrate Specificity Tumor Cells, Cultured
Chemicals
Cyclin E Phosphoproteins RNA Precursors RNA Splicing Factors Ribonucleoprotein, U2 Small Nuclear SF3B1 protein, human Protein Serine-Threonine Kinases CDC2-CDC28 Kinases CDK2 protein, human Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Seghezzi W
Department of Cell Signaling, DNAX Research Institute, Palo Alto, California 94304-1104, USA. seghezzi@dnax.org
Chua K
Shanahan F
Gozani O
Reed R
Lees E
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.18.8.4526
Email
seghezzi@dnax.org
Published
1998-08-00
Pages
4526-36
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC109038
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.18.8.4526 PMC Full Text View journal details
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