Published 17 March 2003. doi:10.1083/jcb.200209097
© The Rockefeller University Press,
0021-9525/2003/3/857 $5.00
The Journal of Cell Biology, Volume 160, Number 6, 857-874
Budding yeast PAK kinases regulate mitotic exit by two different mechanisms
Elena Chiroli,
Roberta Fraschini,
Alessia Beretta,
Mariagrazia Tonelli,
Giovanna Lucchini and
Simonetta Piatti
Dipartimento di Biotecnologie e Bioscienze, Piazza della Scienza 2, 20126 Milano, Italy
Address correspondence to Simonetta Piatti, Dipartimento di Biotecnologie e Bioscienze, Piazza della Scienza 2, 20126 Milano, Italy. Tel.: 39-02-64483547. Fax: 39-02-64483565. E-mail: simonetta.piatti{at}unimib.it
We report the characterization of the dominant-negative CLA4t allele of the budding yeast CLA4 gene, encoding a member of the p21-activated kinase (PAK) family of protein kinases, which, together with its homologue STE20, plays an essential role in promoting budding and cytokinesis. Overproduction of the Cla4t protein likely inhibits both endogenous Cla4 and Ste20 and causes a delay in the onset of anaphase that correlates with inactivation of Cdc20/anaphase-promoting complex (APC)dependent proteolysis of both the cyclinB Clb2 and securin. Although the precise mechanism of APC inhibition by Cla4t remains to be elucidated, our results suggest that Cla4 and Ste20 may regulate the first wave of cyclinB proteolysis mediated by Cdc20/APC, which has been shown to be crucial for activation of the mitotic exit network (MEN). We show that the Cdk1-inhibitory kinase Swe1 is required for the Cla4t-dependent delay in cell cycle progression, suggesting that it might be required to prevent full Cdc20/APC and MEN activation. In addition, inhibition of PAK kinases by Cla4t prevents mitotic exit also by a Swe1-independent mechanism impinging directly on the MEN activator Tem1.
Key Words: cytokinesis; mitotic exit network; Cla4; spindle checkpoint; Swe1
E. Chiroli and R. Fraschini contributed equally to this paper.
The online version of this article includes supplemental material.
* Abbreviations used in this paper: APC, anaphase-promoting complex; GAP, GTPase-activating protein; MEN, mitotic exit network; PAK, p21-activated kinase.

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