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A more recent version of this article appeared on December 1, 2003 Originally published as MBC in Press, 10.1091/mbc.E03-06-0426 on September 5, 2003
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Submitted on June 20, 2003
Revised on July 25, 2003
Accepted on July 28, 2003
1 Departments of Biology and Cell Biology, University Virginia, Charlottesville, VA 22904
2 Department of Molecular Genetics, The Ohio State University, Columbus, OH 43210
3 Banting and Best Department of Medical Research, University of Toronto, Toronto, ON, Canada M5G 1L6,
Department of Medical Genetics and Microbiology,
University of Toronto, Toronto, ON, Canada M5S 1A8
* Corresponding author. E-mail address: park.294{at}osu.edu.
Polarized cell growth requires the coupling of a defined spatial site on the cell cortex to the apparatus that directs the establishment of cell polarity. In the budding yeast S. cerevisiae, the Ras-family GTPase Rsr1p/Bud1p and its regulators select the proper site for bud emergence on the cell cortex. The Rho-family GTPase Cdc42p and its associated proteins then establish an axis of polarized growth by triggering an asymmetric organization of the actin cytoskeleton and secretory apparatus at the selected bud site. We explored whether a direct linkage exists between the Rsr1p/Bud1p and Cdc42p GTPases. Here we show specific genetic interactions between RSR1/BUD1 and particular cdc42 mutants defective in polarity establishment. We also show that Cdc42p coimmunoprecipitated with Rsr1p/Bud1p from yeast extracts. In vitro studies indicated a direct interaction between Rsr1p/Bud1p and Cdc42p, which was enhanced by Cdc24p, a guanine nucleotide exchange factor for Cdc42p. Our findings suggest that Cdc42p interacts directly with Rsr1p/Bud1p in vivo, providing a novel mechanism by which direct contact between a Ras-family GTPase and a Rho-family GTPase links the selection of a growth site to polarity establishment.
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