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Originally published as MBC in Press, 10.1091/mbc.E03-06-0384 on September 5, 2003

Vol. 14, Issue 11, 4592-4604, November 2003

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Genetic and Biochemical Evaluation of the Importance of Cdc6 in Regulating Mitotic Exit

Vincent Archambault, Caihong X. Li, Alan J. Tackett, Ralph Wäsch *, Brian T. Chait, Michael P. Rout, and Frederick R. Cross {dagger}

The Rockefeller University, New York, New York 10021

Submitted June 10, 2003; Revised July 18, 2003; Accepted July 18, 2003
Monitoring Editor: Mark Solomon

We evaluated the hypothesis that the N-terminal region of the replication control protein Cdc6 acts as an inhibitor of cyclin-dependent kinase (Cdk) activity, promoting mitotic exit. Cdc6 accumulation is restricted to the period from mid-cell cycle until the succeeding G1, due to proteolytic control that requires the Cdc6 N-terminal region. During late mitosis, Cdc6 is present at levels comparable with Sic1 and binds specifically to the mitotic cyclin Clb2. Moderate overexpression of Cdc6 promotes viability of CLB2{Delta}db strains, which otherwise arrest at mitotic exit, and rescue is dependent on the N-terminal putative Cdk-inhibitory domain. These observations support the potential for Cdc6 to inhibit Clb2-Cdk, thus promoting mitotic exit. Consistent with this idea, we observed a cytokinesis defect in cdh1{Delta} sic1{Delta} cdc6{Delta}2–49 triple mutants. However, we were able to construct viable strains, in three different backgrounds, containing neither SIC1 nor the Cdc6 Cdk-inhibitory domain, in contradiction to previous work. We conclude, therefore, that although both Cdc6 and Sic1 have the potential to facilitate mitotic exit by inhibiting Clb2-Cdk, mitotic exit nevertheless does not require any identified stoichiometric inhibitor of Cdk activity.

Online version of this article contains supplemental data for some figures. Online version is available at www.molbiolcell.org.

* Present address: Department of Hematology/Oncology, University Medical Center, 79106 Freiburg Germany.

{dagger} Corresponding author. E-mail address: fcross{at}mail.rockefeller.edu.




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