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Switch-domain mutations in the Saccharomycescerevisiae G protein α-subunit Gpa1p identify a receptor subtype-biased mating defect

Abstract

The response to pheromone in Saccharomyces cerevisiae involves a heterotrimeric G protein composed of Gpa1p (α subunit), Ste4p (β) and Ste18p (γ). The switch II region of Gα subunits is involved in several protein-protein interactions and an intrinsic GTPase activity. To investigate the role of this region of Gpa1p, we have analyzed the effect of switch II mutations. The Q323 analog in Gα subunits and Ras is implicated in GTP hydrolysis. Mutation of the Q323 residue of Gpa1p resulted in constitutive activation of the pheromone response pathway and eliminated the ability to interact with Ste4p, consistent with a defect in GTPase activity. Mutation of residue A59 of Ras and the analogous Gαs residue has had quite different effects. The analogous Gpa1p G321T mutation resulted in phenotypes consistent with a less severe GTPase defect, but also led to an unexpected mating phenotype: mating was decreased in both mating types, but the defect was 1000-fold more severe in α cells than in a cells. In addition the G321T mutation resulted in an unusual pheromone response phenotype. We discuss the possibility that these phenotypes may reflect a differential role for the switch II region in activation by the a- and α-factor receptors.

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Received: 5 June 1997 / Accepted: 21 October 1997

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DeSimone, S., Kurjan, J. Switch-domain mutations in the Saccharomycescerevisiae G protein α-subunit Gpa1p identify a receptor subtype-biased mating defect. Mol Gen Genet 257, 662–671 (1998). https://doi.org/10.1007/s004380050695

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  • Key words Pheromone response
  • Mating
  • G proteins