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Eukaryotic Cell, February 2006, p. 330-346, Vol. 5, No. 2
1535-9778/06/$08.00+0     doi:10.1128/EC.5.2.330-346.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Genome-Scale Analysis Reveals Sst2 as the Principal Regulator of Mating Pheromone Signaling in the Yeast Saccharomyces cerevisiae{dagger}

Scott A. Chasse,1 Paul Flanary,1 Stephen C. Parnell,1 Nan Hao,1 Jiyoung Y. Cha,2 David P. Siderovski,2 and Henrik G. Dohlman1,2*

Departments of Biochemistry and Biophysics,1 Pharmacology, University of North Carolina, Chapel Hill, North Carolina 27599-72602

Received 21 November 2005/ Accepted 13 December 2005

A common property of G protein-coupled receptors is that they become less responsive with prolonged stimulation. Regulators of G protein signaling (RGS proteins) are well known to accelerate G protein GTPase activity and do so by stabilizing the transition state conformation of the G protein {alpha} subunit. In the yeast Saccharomyces cerevisiae there are four RGS-homologous proteins (Sst2, Rgs2, Rax1, and Mdm1) and two G{alpha} proteins (Gpa1 and Gpa2). We show that Sst2 is the only RGS protein that binds selectively to the transition state conformation of Gpa1. The other RGS proteins also bind Gpa1 and modulate pheromone signaling, but to a lesser extent and in a manner clearly distinct from Sst2. To identify other candidate pathway regulators, we compared pheromone responses in 4,349 gene deletion mutants representing nearly all nonessential genes in yeast. A number of mutants produced an increase (sst2, bar1, asc1, and ygl024w) or decrease (cla4) in pheromone sensitivity or resulted in pheromone-independent signaling (sst2, pbs2, gas1, and ygl024w). These findings suggest that Sst2 is the principal regulator of Gpa1-mediated signaling in vivo but that other proteins also contribute in distinct ways to pathway regulation.

* Corresponding author. Mailing address: Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599-7260. Phone: (919) 843-6894. Fax: (919) 966-2852. E-mail: hdohlman{at}med.unc.edu.

{dagger} Supplemental material for this article may be found at http://ec.asm.org/.

Eukaryotic Cell, February 2006, p. 330-346, Vol. 5, No. 2
1535-9778/06/$08.00+0     doi:10.1128/EC.5.2.330-346.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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