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21 (15): 4012-4025

Copyright © 2002 by the European Molecular Biology Organization.

The Rho-GAP Bem2p plays a GAP-independent role in the morphogenesis checkpoint

Aron R. Marquitz, Jacob C. Harrison, Indrani Bose1, Trevin R. Zyla, John N. McMillan, and Daniel J. Lew2

Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, NC 27710, USA 1 Present address: Department of Molecular Microbiology, Washington University School of Medicine, St Louis, MO 63110, USA 2 Corresponding author e-mail: daniel.lew{at}duke.edu

Abstract: The Saccharomyces cerevisiae morphogenesis checkpoint delays mitosis in response to insults that impair actin organization and/or bud formation. The delay is due to accumulation of the inhibitory kinase Swe1p, which phosphorylates the cyclin-dependent kinase Cdc28p. Having screened through a panel of yeast mutants with defects in cell morphogenesis, we report here that the polarity establishment protein Bem2p is required for the checkpoint response. Bem2p is a Rho-GTPase activating protein (GAP) previously shown to act on Rho1p, and we now show that it also acts on Cdc42p, the GTPase primarily responsible for establishment of cell polarity in yeast. Whereas the morphogenesis role of Bem2p required GAP activity, the checkpoint role of Bem2p did not. Instead, this function required an N-terminal Bem2p domain. Thus, this single protein has a GAP-dependent role in promoting cell polarity and a GAP-independent role in responding to defects in cell polarity by enacting the checkpoint. Surprisingly, Swe1p accumulation occurred normally in bem2 cells, but they were nevertheless unable to promote Cdc28p phosphorylation. Therefore, Bem2p defines a novel pathway in the morphogenesis checkpoint.

Key Words: Keywords: Bem2p/cell cycle/checkpoint/Rho-GAP/Saccharomyces cerevisiae

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