Cyclin A-dependent phosphorylation of the ETS-related protein, MEF, restricts its activity to the G1 phase of the cell cycle Journal Article
| Authors: | Miyazaki, Y.; Boccuni, P.; Mao, S.; Zhang, J.; Erdjument-Bromage, H.; Tempst, P.; Kiyokawa, H.; Nimer, S. D. |
| Article Title: | Cyclin A-dependent phosphorylation of the ETS-related protein, MEF, restricts its activity to the G1 phase of the cell cycle |
| Abstract: | MEF, a recently identified member of the E74 family of ETS-related transcription factors, is a strong transcriptional activator of cytokine gene expression. Using a green fluorescent protein gene reporter plasmid regulated by an MEF-responsive promoter, we determined that the transcriptional activity of MEF is largely restricted to the G1 phase of the cell cycle. MEF-dependent transcription was suppressed by the expression of cyclin A but not by cyclin D or cyclin E. This effect was due to the kinase activity generated by cyclin A expression, as co-expression of the cyclin-dependent kinase inhibitors p21 or p27, or a dominant negative form of CDK2 (DNK2), abrogated the reduction of MEF transcriptional activity by cyclin A. Cyclin A-CDK2 phosphorylated MEF protein in vitro more efficiently than cyclin D-CDK4 or cyclin E-CDK2, and phosphorylation of MEF by cyclin A-CDK2 reduced its ability to bind DNA. We determined one site of phosphorylation by cyclin A-CDK2 at the C terminus of MEF, using mass-spectrometry; mutation of three serine or threonine residues in this region significantly reduced phosphorylation of MEF by cyclin A and reduced cyclin A-mediated suppression of its transactivating activity. These amino acid substitutions also reduced the restriction of MEF activity to G1. Phosphorylation of MEF by the cyclin A-CDK2 complex controls its transcriptional activity during the cell cycle, establishing a novel link between the ETS family of proteins and the cell cycle machinery. |
| Keywords: | controlled study; protein phosphorylation; unclassified drug; dna binding protein; mutation; dna-binding proteins; nonhuman; molecular genetics; mass spectrometry; proteins; animal cell; animal; metabolism; animals; genes; gene expression; amino acid substitution; carboxy terminal sequence; protein protein interaction; transcription initiation; green fluorescent protein; transcription factor; genetic transcription; transcription, genetic; enzyme activity; cos cells; phosphorylation; physiology; animalia; transcription factors; cytokine; chemistry; dna; amino acid sequence; molecular sequence data; nucleotide sequence; protein p27; plasmid; cell strain cos1; reporter gene; base sequence; amino acid; threonine; dna primers; primer dna; amino acids; protein family; dna binding; cyclin a; cyclin dependent kinase inhibitor; protein p21; phosphotransferase; mutagenesis; cell cycle g1 phase; g1 phase; cyclin e; cyclin d; serine derivative; priority journal; article; myocyte enhancer factor |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 276 |
| Issue: | 44 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2001-11-02 |
| Start Page: | 40528 |
| End Page: | 40536 |
| Language: | English |
| DOI: | 10.1074/jbc.M103051200 |
| PUBMED: | 11504716 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 21 May 2015 -- Source: Scopus |
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