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Mol. Cell. Biol. 20 (23): 8826-8835
Copyright © 2000 by the American Society for Microbiology. All rights reserved.
Molecular and Cellular Biology, December 2000, p. 8826-8835, Vol. 20, No. 23
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Localization and Signaling of G
Subunit Ste4p Are Controlled by a-Factor Receptor and the
a-Specific Protein Asg7p
Jinah
Kim,1
Eric
Bortz,1
Hualin
Zhong,2,
Thomas
Leeuw,3,
Ekkehard
Leberer,3,
Andrew K.
Vershon,2 and
Jeanne
P.
Hirsch1,*
Department of Cell Biology and Anatomy, Mount
Sinai School of Medicine, New York, New York
100291; Waksman Institute and
Department of Molecular Biology and Biochemistry, Rutgers University,
Piscataway, New Jersey 088542; and
Eukaryotic Genetics Group, Biotechnology Research
Institute, National Research Council of Canada, Montreal, Quebec
H4P 2R2, Canada3
Received 29 June 2000/Returned for modification 22 August
2000/Accepted 15 September 2000
Haploid yeast cells initiate pheromone signaling upon the binding
of pheromone to its receptor and activation of the coupled G protein. A
regulatory process termed receptor inhibition blocks pheromone
signaling when the a-factor receptor is inappropriately expressed in
MATa cells. Receptor inhibition blocks signaling by
inhibiting the activity of the G protein  subunit, Ste4p. To
investigate how Ste4p activity is inhibited, its subcellular location
was examined. In wild-type cells,  -factor treatment resulted in
localization of Ste4p to the plasma membrane of mating projections. In
cells expressing the a-factor receptor, -factor treatment resulted
in localization of Ste4p away from the plasma membrane to an internal
compartment. An altered version of Ste4p that is largely insensitive to
receptor inhibition retained its association with the membrane in cells
expressing the a-factor receptor. The inhibitory function of the
a-factor receptor required ASG7, an a-specific gene of
previously unknown function. ASG7 RNA was induced by
pheromone, consistent with increased inhibition as the pheromone
response progresses. The a-factor receptor inhibited signaling in its
liganded state, demonstrating that the receptor can block the signal
that it initiates. ASG7 was required for the altered
localization of Ste4p that occurs during receptor inhibition, and the
subcellular location of Asg7p was consistent with its having a direct
effect on Ste4p localization. These results demonstrate that Asg7p
mediates a regulatory process that blocks signaling from a G protein
subunit and causes its relocalization within the cell.
*
Corresponding author. Mailing address: Box 1007, Mount
Sinai School of Medicine, 1 Gustave Levy Place, New York, NY 10029. Phone: (212) 241-0224. Fax: (212) 860-1174. E-mail:
Jeanne.Hirsch{at}.mssm.edu.
Present address: Laboratory of Cell Biology, Howard Hughes Medical
Institute, The Rockefeller University, New York, NY 10021.
Present address: Aventis Biotechnologie, D-82152 Martinsried, Germany.
Molecular and Cellular Biology, December 2000, p. 8826-8835, Vol. 20, No. 23
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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