Effects of Expression of Mammalian GeL and Hybrid Mammalian-Yeast Ga Proteins on the Yeast Pheromone Response Signal Transduction Pathway

Scgl, the product of the Saccharomyces cerevisiae SCGI (also called GPAI) gene, is homologous to the a subunits of G proteins involved in signal transduction in mammalian cells. Scgl negatively controls the pheromone response pathway in haploid cells. Either pheromonal activation or an scgl null mutation relieves the negative control and leads to an arrest of cell growth in the Gl phase of the cell cycle. Expression of rat Gas was previously shown to complement the growth defect of scgl null mutants while not allowing mating. We have extended this analysis to examine the effects of the short form of Gas (which lacks 15 amino acids present in the long form), Gai2, Gao, and Scgl-mammalian Ga hybrids. In addition, we have found that constructs able to complement scgl are also able to inhibit the response to pheromone and mating when expressed in a wild-type SCGI strain. Overexpression of Scgl has a similar inhibitory effect. These results are consistent with a model proposed for the action of Scgl as the a component of a heterotrimeric G protein in which the fry component (Ste/Stel8) activates the pheromone response after dissociation from Scgl. They suggest that the Ga constructs able to complement scgl can interact with ,y to prevent activation ofthe pathway but are unable to interact with pheromone receptors to activate the pathway. The G protein family transduces hormonal and sensory signals

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