![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
Vol. 11, Issue 3, 957-968, March 2000
and
G
Subunits Are Required for Full Activity of the
Pheromone Response Pathway in Saccharomyces cerevisiae
Department of Cell Biology and Physiology, Washington University
School of Medicine, St. Louis, Missouri 63110
To establish the biological function of thioacylation
(palmitoylation), we have studied the heterotrimeric guanine
nucleotide-binding protein (G protein) subunits of the
pheromone response pathway of Saccharomyces
cerevisiae. The yeast G protein 
subunit (Ste18p) is
unusual among G
subunits because it is farnesylated at cysteine 107 and has the potential to be thioacylated at cysteine 106. Substitution of either cysteine results in a strong signaling defect.
In this study, we found that Ste18p is thioacylated at cysteine 106, which depended on prenylation of cysteine 107. Ste18p was targeted to
the plasma membrane even in the absence of prenylation or
thioacylation. However, G protein activation released prenylation- or
thioacylation-defective Ste18p into the cytoplasm. Hence, lipid modifications of the G subunit are dispensable for G
protein activation by receptor, but they are required to maintain the plasma membrane association of G
after
receptor-stimulated release from G
. The G protein 
subunit (Gpa1p) is tandemly modified at its N terminus with amide- and
thioester-linked fatty acids. Here we show that Gpa1p was thioacylated
in vivo with a mixture of radioactive myristate and palmitate. Mutation
of the thioacylation site in Gpa1p resulted in yeast cells that
displayed partial activation of the pathway in the absence of
pheromone. Thus, dual lipidation motifs on Gpa1p and Ste18p are
required for a fully functional pheromone response pathway.
This article has been cited by other articles:
|
|
 |
|
  M. P. Torres, M. J. Lee, F. Ding, C. Purbeck, B. Kuhlman, N. V. Dokholyan, and H. G. Dohlman G Protein Mono-ubiquitination by the Rsp5 Ubiquitin Ligase J. Biol. Chem., March 27, 2009; 284(13): 8940 - 8950. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Trusov, W. Zhang, S. M. Assmann, and J. R. Botella G{gamma}1 + G{gamma}2 != G{beta}: Heterotrimeric G Protein G{gamma}-Deficient Mutants Do Not Recapitulate All Phenotypes of G{beta}-Deficient Mutants Plant Physiology, June 1, 2008; 147(2): 636 - 649. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. D. Slaughter, J. W. Schwartz, and R. Li Mapping dynamic protein interactions in MAP kinase signaling using live-cell fluorescence fluctuation spectroscopy and imaging PNAS, December 18, 2007; 104(51): 20320 - 20325. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. A. Johnston and D. P. Siderovski Receptor-Mediated Activation of Heterotrimeric G-Proteins: Current Structural Insights Mol. Pharmacol., August 1, 2007; 72(2): 219 - 230. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. Zeng, X. Wang, and M. P. Running Dual Lipid Modification of Arabidopsis G{gamma}-Subunits Is Required for Efficient Plasma Membrane Targeting Plant Physiology, March 1, 2007; 143(3): 1119 - 1131. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. J. W. Adjobo-Hermans, J. Goedhart, and T. W. J. Gadella Jr Plant G protein heterotrimers require dual lipidation motifs of G{alpha} and G{gamma} and do not dissociate upon activation J. Cell Sci., December 15, 2006; 119(24): 5087 - 5097. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Wang and R. J. Deschenes Plasma Membrane Localization of Ras Requires Class C Vps Proteins and Functional Mitochondria in Saccharomyces cerevisiae Mol. Cell. Biol., April 15, 2006; 26(8): 3243 - 3255. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. G J M Kuiper, W. Klootwijk, and T. J Visser Expression of recombinant membrane-bound type I iodothyronine deiodinase in yeast J. Mol. Endocrinol., June 1, 2005; 34(3): 865 - 878. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. A. Gubitosi-Klug, D. J. Mancuso, and R. W. Gross The human Kv1.1 channel is palmitoylated, modulating voltage sensing: Identification of a palmitoylation consensus sequence PNAS, April 26, 2005; 102(17): 5964 - 5968. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Cherukuri, R. H. Carter, S. Brooks, W. Bornmann, R. Finn, C. S. Dowd, and S. K. Pierce B Cell Signaling Is Regulated by Induced Palmitoylation of CD81 J. Biol. Chem., July 23, 2004; 279(30): 31973 - 31982. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. R. Hynes, L. Tang, S. M. Mervine, J. L. Sabo, E. A. Yost, P. N. Devreotes, and C. H. Berlot Visualization of G Protein {beta}{gamma} Dimers Using Bimolecular Fluorescence Complementation Demonstrates Roles for Both {beta} and {gamma} in Subcellular Targeting J. Biol. Chem., July 16, 2004; 279(29): 30279 - 30286. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Gettemans, K. Meerschaert, J. Vandekerckhove, and V. De Corte A Kelch {beta} Propeller Featuring as a G{beta} Structural Mimic: Reinventing the Wheel? Sci. Signal., July 15, 2003; 2003(191): pe27 - pe27. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. L. Chinault and K. J. Blumer The C-terminal Tail Preceding the CAAX Box of a Yeast G Protein {gamma} Subunit Is Dispensable for Receptor-mediated G Protein Activation in Vivo J. Biol. Chem., May 30, 2003; 278(23): 20638 - 20644. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Takida and P. B. Wedegaertner Heterotrimer Formation, Together with Isoprenylation, Is Required for Plasma Membrane Targeting of Gbeta gamma J. Biol. Chem., May 2, 2003; 278(19): 17284 - 17290. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. A. Duncan and A. G. Gilman Characterization of Saccharomyces cerevisiae Acyl-protein Thioesterase 1, the Enzyme Responsible for G Protein alpha Subunit Deacylation in Vivo J. Biol. Chem., August 23, 2002; 277(35): 31740 - 31752. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. J. Duan, I. Xenarios, and D. Eisenberg Describing Biological Protein Interactions in Terms of Protein States and State Transitions : THE LiveDIP DATABASE Mol. Cell. Proteomics, February 1, 2002; 1(2): 104 - 116. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Sette, C. J. Inouye, S. L. Stroschein, P. J. Iaquinta, and J. Thorner Mutational Analysis Suggests That Activation of the Yeast Pheromone Response Mitogen-activated Protein Kinase Pathway Involves Conformational Changes in the Ste5 Scaffold Protein Mol. Biol. Cell, November 1, 2000; 11(11): 4033 - 4049. [Abstract] [Full Text]
|
|
