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J. Biol. Chem., Vol. 277, Issue 30, 26721-26724, July 26, 2002

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ACCELERATED PUBLICATION
Localization of the Rsr1/Bud1 GTPase Involved in Selection of a Proper Growth Site in Yeast^*,

Hay-Oak Park^§¶, Pil Jung Kang, and Amy Wilson Rachfal^§

From the  Department of Molecular Genetics and ^§ Graduate Program in Molecular, Cellular, and Developmental Biology, The Ohio State University, Columbus, Ohio 43210-1292

Yeast cells organize their actin cytoskeleton in a highly polarized manner during vegetative growth. The Ras-like GTPase Rsr1/Bud1 and its regulators are required for selection of a specific site for growth. Here we showed that Rsr1/Bud1 was broadly distributed on the plasma membrane and highly concentrated at the incipient bud site and polarized growth sites. We also showed that localization of Cdc24, a guanine nucleotide exchange factor for the Cdc42 GTPase, to the proper bud site was dependent on Rsr1/Bud1. Surprisingly, Rsr1/Bud1 also localized to intracellular membranes. A mutation in the lysine repeat in the hypervariable region of Rsr1/Bud1 specifically abolished its plasma membrane localization, whereas a mutation at the CAAX motif eliminated both plasma membrane and internal membrane association of Rsr1/Bud1. Thus the lysine repeat and the CAAX motif of Rsr1/Bud1 are important for its localization to the plasma membrane and to the polarized growth sites. This localization of Rsr1/Bud1 is essential for its function in proper bud site selection because both mutations resulted in random bud site selection.

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