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J. Biol. Chem., Vol. 278, Issue 24, 21798-21804, June 13, 2003

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G Recruits Rho1 to the Site of Polarized Growth during Mating in Budding Yeast^*

Eli E. Bar, Alexis T. Ellicott and David E. Stone 

From the Department of Biological Sciences, Laboratory for Molecular Biology, University of Illinois at Chicago, Chicago, Illinois 60607

In mating mixtures of Saccharomyces cerevisiae, cells polarize their growth toward their conjugation partners along a pheromone gradient. This chemotropic phenomenon is mediated by structural proteins such as Far1 and Bem1 and by signaling proteins such as Cdc24, Cdc42, and G. The G subunit is thought to provide a positional cue that recruits the polarity establishment proteins, and thereby induces polarization of the actin cytoskeleton. We identified RHO1 in a screen for allelespecific high-copy suppressors of G overexpression, suggesting that Rho1 binds G in vivo. Inactivation of Rho1 GTPase activity augmented the rescue phenotype, suggesting that it is the activated form of Rho1 that binds G. We also found, in a pull-down assay, that Rho1 associates with GST-Ste4 and that Rho1 is localized to the neck and tip of mating projections. Moreover, a mutation in STE4 that disrupts G-Rho1 interaction reduces the projection tip localization of Rho1 and compromises the integrity of pheromone-treated cells deficient in Rho1 activity. In addition to its roles as a positive regulator of 1,3--glucan synthase and of the cell integrity MAP kinase cascade, it was recently shown that Rho1 is necessary for the formation of mating projections. Together, these results suggest that G recruits Rho1 to the site of polarized growth during mating.

Received for publication, December 11, 2002 , and in revised form, March 20, 2003.

^* This work was supported by American Cancer Society Research Grant RPG-94-034-06-MBC and by National Science Foundation Grant MCB-0111 397. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed. Tel.: 312-996-5710; Fax: 312-413-2691; E-mail: dstone{at}uic.edu.

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