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Molecular and Cellular Biology, August 2000, p. 5858-5864, Vol. 20, No. 16
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Cks1 Is Required for G1 Cyclin-Cyclin-Dependent Kinase Activity in Budding Yeast

Gregory J. Reynard, William Reynolds, Rati Verma, and Raymond J. Deshaies*

Division of Biology, California Institute of Technology, Pasadena, California 91125

Received 17 March 2000/Returned for modification 26 April 2000/Accepted 19 May 2000

p13suc1 (Cks) proteins have been implicated in the regulation of cyclin-dependent kinase (CDK) activity. However, the mechanism by which Cks influences the function of cyclin-CDK complexes has remained elusive. We show here that Cks1 is required for the protein kinase activity of budding yeast G1 cyclin-CDK complexes. Cln2 and Cdc28 subunits coexpressed in baculovirus-infected insect cells fail to exhibit protein kinase activity towards multiple substrates in the absence of Cks1. Cks1 can both stabilize Cln2-Cdc28 complexes and activate intact complexes in vitro, suggesting that it plays multiple roles in the biogenesis of active G1 cyclin-CDK complexes. In contrast, Cdc28 forms stable, active complexes with the B-type cyclins Clb4 and Clb5 regardless of whether Cks1 is present. The levels of Cln2-Cdc28 and Cln3-Cdc28 protein kinase activity are severely reduced in cks1-38 cell extracts. Moreover, phosphorylation of G1 cyclins, which depends on Cdc28 activity, is reduced in cks1-38 cells. The role of Cks1 in promoting G1 cyclin-CDK protein kinase activity both in vitro and in vivo provides a simple molecular rationale for the essential role of CKS1 in progression through G1 phase in budding yeast.

* Corresponding author. Mailing address: Division of Biology, 156-29, California Institute of Technology, Pasadena, CA 91125. Phone: (626) 395-3162. Fax: (626) 449-0756. E-mail: deshaies{at}its.caltech.edu.

Molecular and Cellular Biology, August 2000, p. 5858-5864, Vol. 20, No. 16
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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