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Genetic screens in yeast to identify mammalian nonreceptor modulators of G-protein signaling

Nature Biotechnology volume 17, pages 878–883 (1999)Cite this article

Abstract

We describe genetic screens in Saccharomyces cerevisiae designed to identify mammalian nonreceptor modulators of G-protein signaling pathways. Strains lacking a pheromone-responsive G-protein coupled receptor and expressing a mammalian-yeast Gα hybrid protein were made conditional for growth upon either pheromone pathway activation (activator screen) or pheromone pathway inactivation (inhibitor screen). Mammalian cDNAs that conferred plasmid-dependent growth under restrictive conditions were identified. One of the cDNAs identified from the activator screen, a human Ras-related G protein that we term AGS1 (for activator of G-protein signaling), appears to function by facilitating guanosine triphosphate (GTP) exchange on the heterotrimeric Gα. A cDNA product identified from the inhibitor screen encodes a previously identified regulator of G-protein signaling, human RGS5.

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Figure 1: Engineering of the yeast pheromone response pathway.
Figure 2: Characterization of mammalian modulators of the yeast pheromone pathway.
Figure 3: Characterization of clone L1 (AGS1).
Figure 4: Gα selectivity profile of AGS1.

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Acknowledgements

We thank Drs. Elliott Ross, Henry Bourne, Josh Trueheart, and Ben Benton for helpful discussions and comments, Gary Meissner, Ralph Vaccaro, and Jin Xie for their technical assistance, and Drs. George Sprague, Susan Michaelis, Roger Perlmuter, Mel Simon, Janet Kurjan, and David Stone for providing materials used in this study. S.M.L. was supported by the National Institutes of Health grant RO1-NS24821. A.T. was a visiting graduate student from the Department of Pharmaceutical Sciences, University of Tokyo, Japan, and was supported in part by a University of Tokyo scholarship.

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  1. Cadus Pharmaceutical Corporation, 777 Old Saw Mill River Rd., Tarrytown, 10591, NY

    Mary J. Cismowski, Chienling Ma, Jeffrey S. Lizano, Xiaobing Xie, Hans Fuernkranz & Emir Duzic

  2. Department of Pharmacology, Medical University of South Carolina, 171 Ashley Ave., Charleston, 29425, SC

    Aya Takesono & Stephen M. Lanier

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Correspondence to Emir Duzic.

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Cismowski, M., Takesono, A., Ma, C. et al. Genetic screens in yeast to identify mammalian nonreceptor modulators of G-protein signaling. Nat Biotechnol 17, 878–883 (1999). https://doi.org/10.1038/12867

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