Requirements for CDK7 in the assembly of CDK1/cyclin B and activation of CDK2 revealed by chemical genetics in human cells Journal Article

   

Authors:	Larochelle, S.; Merrick, K. A.; Terret, M. E.; Wohlbold, L.; Barboza, N. M.; Zhang, C.; Shokat, K. M.; Jallepalli, P. V.; Fisher, R. P.
Article Title:	Requirements for CDK7 in the assembly of CDK1/cyclin B and activation of CDK2 revealed by chemical genetics in human cells
Abstract:	Cell division is controlled by cyclin-dependent kinases (CDKs). In metazoans, S phase onset coincides with activation of Cdk2, whereas Cdk1 triggers mitosis. Both Cdk1 and -2 require cyclin binding and T loop phosphorylation for full activity. The only known CDK-activating kinase (CAK) in metazoans is Cdk7, which is also part of the transcription machinery. To test the requirements for Cdk7 in vivo, we replaced wild-type Cdk7 with a version sensitive to bulky ATP analogs in human cancer cells. Selective inhibition of Cdk7 in G1 prevents activation (but not formation) of Cdk2/cyclin complexes and delays S phase. Inhibiting Cdk7 in G2 blocks entry to mitosis and disrupts Cdk1/cyclin B complex assembly, indicating that the two steps of Cdk1 activation-cyclin binding and T loop phosphorylation-are mutually dependent. Therefore, by combining chemical genetics and homologous gene replacement in somatic cells, we reveal different modes of CDK activation by Cdk7 at two distinct execution points in the cell cycle. © 2007 Elsevier Inc. All rights reserved.
Keywords:	controlled study; protein phosphorylation; human cell; protein function; mitosis; homologous recombination; cell survival; cell cycle; cell cycle progression; cell cycle s phase; cell division; complex formation; enzyme inhibition; protein assembly; protein protein interaction; enzyme activation; enzyme regulation; transcription regulation; colon cancer; enzyme inactivation; cell cycle arrest; chemical genetics; cellcycle; cell cycle g2 phase; adenosine triphosphate; cyclin-dependent kinases; cyclin b; cell cycle regulation; enzyme binding; cell cycle g1 phase; s phase; enzyme mechanism; cyclin dependent kinase 1; cyclin dependent kinase 7; cyclin dependent kinase activating kinase; metazoa; cyclin dependent kinase 2; cyclin-dependent kinase 2; cdc2 protein kinase
Journal Title:	Molecular Cell
Volume:	25
Issue:	6
ISSN:	1097-2765
Publisher:	Cell Press
Date Published:	2007-03-23
Start Page:	839
End Page:	850
Language:	English
DOI:	10.1016/j.molcel.2007.02.003
PUBMED:	17386261
PROVIDER:	scopus
PMCID:	PMC1858677
DOI/URL:	http://www.scopus.com/inward/record.url?eid=2-s2.0-33947424005&partnerID=40&md5=bf6eb1e93eebb475cab58a6d46e8f46f
Notes:	--- - "Cited By (since 1996): 59" - "Export Date: 17 November 2011" - "CODEN: MOCEF" - "Source: Scopus"

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