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© The Rockefeller University Press, 0021-9525/1999//1187 $5.00
The Journal of Cell Biology, Volume 144, Number 6, , 1999 1187-1202

A Cdc24p-Far1p-Gβ Protein Complex Required for Yeast Orientation during Mating

Division of Cell Biology, MRC Laboratory of Molecular Biology, Cambridge, CB2 2QH, United Kingdom

Oriented cell growth requires the specification of a site for polarized growth and subsequent orientation of the cytoskeleton towards this site. During mating, haploid Saccharomyces cerevisiae cells orient their growth in response to a pheromone gradient overriding an internal landmark for polarized growth, the bud site. This response requires Cdc24p, Far1p, and a heterotrimeric G-protein. Here we show that a two- hybrid interaction between Cdc24p and Gβ requires Far1p but not pheromone-dependent MAP-kinase signaling, indicating Far1p has a role in regulating the association of Cdc24p and Gβ. Binding experiments demonstrate that Cdc24p, Far1p, and Gβ form a complex in which pairwise interactions can occur in the absence of the third protein. Cdc24p localizes to sites of polarized growth suggesting that this complex is localized. In the absence of CDC24-FAR1-mediated chemotropism, a bud site selection protein, Bud1p/Rsr1p, is essential for morphological changes in response to pheromone. These results suggest that formation of a Cdc24p-Far1p-Gβ complex functions as a landmark for orientation of the cytoskeleton during growth towards an external signal.

Key Words: chemotropism • landmark • oriented growth • Ste4p Ste18p • yeast mating

Abbreviations used in this paper: GFP, green fluorescent protein; GST, glutathione-S-transferase; HA, hemagglutinin; MAP, mitogen-activated protein; MBP, maltose binding protein; TPI, triose phosphate isomerase; TEV, tobacco etch virus.

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