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Molecular and Cellular Biology, July 2000, p. 4483-4493, Vol. 20, No. 13
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Testing Cyclin Specificity in the Exit from Mitosis

Matthew D. Jacobson,1 Samantha Gray,2 Maria Yuste-Rojas,3 and Frederick R. Cross1,*

The Rockefeller University1 and Boston Consulting Group,2 New York, New York, and Pharma-Mar, Madrid, Spain3

Received 18 January 2000/Returned for modification 10 March 2000/Accepted 14 April 2000

Cyclical inactivation of B-type cyclins has been proposed to be required for alternating DNA replication and mitosis. Destruction box-dependent Clb5p degradation is strongly increased in mitotic cells, and constitutive overexpression of Clb5p lacking the destruction box resulted in rapid accumulation of inviable cells, frequently multiply budded, with DNA contents ranging from unreplicated to apparently fully replicated. Loss of viability correlated with retention of nuclear Clb5p at the time of nuclear division. CLB2-Delta db overexpression that was quantitatively comparable to CLB5-Delta db overexpression with respect to Clb protein production and Clb-associated kinase activity resulted in a distinct phenotype: reversible mitotic arrest with uniformly replicated DNA. Simultaneous overexpression of CLB2-Delta db and CLB5-Delta db overexpressers similarly resulted in a uniform arrest with replicated DNA, and this arrest was significantly more reversible than that observed with CLB5-Delta db overexpression alone. These results suggest that Clb2p and not Clb5p can efficiently block mitotic completion. We speculate that CLB5-Delta db overexpression may be lethal, because persistence of high nuclear Clb5p-associated kinase throughout mitosis leads to failure to load origins of replication, thus preventing DNA replication in the succeeding cell cycle.

* Corresponding author. Mailing address: The Rockefeller University, Box 327, 1230 York Ave., New York, NY 10021. Phone: (212) 327-7685. Fax: (212) 327-7923. E-mail: fcross{at}rockvax.rockefeller.edu.

Molecular and Cellular Biology, July 2000, p. 4483-4493, Vol. 20, No. 13
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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