Genetics, Vol 143, 103-117, Copyright © 1996


INVESTIGATIONS

Genetic Relationships Between the G Protein {beta}{gamma} Complex, Ste5p, Ste20p and Cdc42p: Investigation of Effector Roles in the Yeast Pheromone Response Pathway

R. Akada, L. Kallal, D. I. Johnson and J. Kurjan
Present address: Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University, Ube 755, Japan.

The Saccharomyces cerevisiae G protein {beta}{gamma} dimer, Ste4p/Ste18p, acts downstream of the {alpha} subunit, Gpa1p, to activate the pheromone response pathway and therefore must interact with a downstream effector. Synthetic sterile mutants that exacerbate the phenotype of ste4-ts mutations were isolated to identify proteins that functionally interact with Ste4p. The identification of a ste18 mutant indicated that this screen could identify proteins that interact directly with Ste4p. The other mutations were in STE5 and the STE20 kinase gene, which act near Ste4p in the pathway, and a new gene called STE21. ste20 null mutants showed residual mating, suggesting that another kinase may provide some function. Overexpression of Ste5p under galactose control activated the pheromone response pathway. This activation was dependent on Ste4p and Ste18p and partially dependent on Ste20p. These results cannot be explained by the linear pathway of Ste4p -> Ste20p -> Ste5p. Overexpression of Cdc42p resulted in a slight increase in pheromone induction of a reporter gene, and overexpression of activated forms of Cdc42p resulted in a further twofold increase. Mutations in pheromone response pathway components did not suppress the lethality associated with the activated CDC42 mutations, suggesting that this effect is independent of the pheromone response pathway.


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