Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 5 October 1990:
Vol. 250. no. 4977, pp. 121 - 123
DOI: 10.1126/science.2171146
Prev | Table of Contents | Next

Articles

Science, Vol 250, Issue 4977, 121-123
Copyright © 1990 by American Association for the Advancement of Science

articles

Control of yeast mating signal transduction by a mammalian beta 2-adrenergic receptor and Gs alpha subunit

K King, HG Dohlman, J Thorner, MG Caron, and RJ Lefkowitz

Department of Medicine (Cardiology), Howard Hughes Medical Institute, Duke University Medical Center, Durham, NC 27710.

To facilitate functional and mechanistic studies of receptor-G protein interactions, [corrected] the human beta 2-adrenergic receptor (h beta-AR) has been expressed in Saccharomyces cerevisiae. This was achieved by placing a modified h beta-AR gene under control of the galactose-inducible GAL1 promoter. After induction by galactose, functional h beta-AR was expressed at a concentration several hundred times as great as that found in any human tissue. As determined from competitive ligand binding experiments, h beta-AR expressed in yeast displayed characteristic affinities, specificity, and stereoselectivity. Partial activation of the yeast pheromone response pathway by beta-adrenergic receptor agonists was achieved in cells coexpressing h beta-AR and a mammalian G protein (Gs) alpha subunit-demonstrating that these components can couple to each other and to downstream effectors when expressed in yeast. This in vivo reconstitution system provides a new approach for examining ligand binding and G protein coupling to cell surface receptors.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Designing biological systems.
D. A. Drubin, J. C. Way, and P. A. Silver (2007)
Genes & Dev. 21, 242-254
   Abstract »    Full Text »    PDF »
Functional Characterization of an {alpha}-Factor-Like Sordaria macrospora Peptide Pheromone and Analysis of Its Interaction with Its Cognate Receptor in Saccharomyces cerevisiae.
S. Mayrhofer and S. Poggeler (2005)
Eukaryot. Cell 4, 661-672
   Abstract »    Full Text »    PDF »
Identification of Allosteric Peptide Agonists of CXCR4.
A. Sachpatzidis, B. K. Benton, J. P. Manfredi, H. Wang, A. Hamilton, H. G. Dohlman, and E. Lolis (2003)
J. Biol. Chem. 278, 896-907
   Abstract »    Full Text »    PDF »
Functional Expression of Heteromeric Calcitonin Gene-related Peptide and Adrenomedullin Receptors in Yeast.
J. J. Miret, L. Rakhilina, L. Silverman, and B. Oehlen (2002)
J. Biol. Chem. 277, 6881-6887
   Abstract »    Full Text »    PDF »
The G Protein-Coupled Receptor Gpr1 Is a Nutrient Sensor That Regulates Pseudohyphal Differentiation in Saccharomyces cerevisiae.
M. C. Lorenz, X. Pan, T. Harashima, M. E. Cardenas, Y. Xue, J. P. Hirsch, and J. Heitman (2000)
Genetics 154, 609-622
   Abstract »    Full Text »
Physiological Regulation of G Protein-Linked Signaling.
A. J. Morris and C. C. Malbon (1999)
Physiol Rev 79, 1373-1430
   Abstract »    Full Text »    PDF »
Multiple Sex Pheromones and Receptors of a Mushroom-producing Fungus Elicit Mating in Yeast.
T. J. Fowler, S. M. DeSimone, M. F. Mitton, J. Kurjan, and C. A. Raper (1999)
Mol. Biol. Cell 10, 2559-2572
   Abstract »    Full Text »
Identification of a Polar Region in Transmembrane Domain 6 That Regulates the Function of the G Protein-Coupled alpha -Factor Receptor.
P. Dube and J. B. Konopka (1998)
Mol. Cell. Biol. 18, 7205-7215
   Abstract »    Full Text »    PDF »
Dominant-Negative Mutations in the G-Protein-Coupled alpha -Factor Receptor Map to the Extracellular Ends of the Transmembrane Segments.
M. Dosil, L. Giot, C. Davis, and J. B. Konopka (1998)
Mol. Cell. Biol. 18, 5981-5991
   Abstract »    Full Text »    PDF »
Afr1p Regulates the Saccharomyces cerevisiae {alpha}-Factor Receptor by a Mechanism That Is Distinct From Receptor Phosphorylation and Endocytosis.
C. Davis, P. Dube, and J. B. Konopka (1998)
Genetics 148, 625-636
   Abstract »    Full Text »    PDF »
Edg-2/Vzg-1 Couples to the Yeast Pheromone Response Pathway Selectively in Response to Lysophosphatidic Acid.
J. R. Erickson, J. J. Wu, J. G. Goddard, G. Tigyi, K. Kawanishi, L. D. Tomei, and M. C. Kiefer (1998)
J. Biol. Chem. 273, 1506-1510
   Abstract »    Full Text »    PDF »
Expression and Purification of the Saccharomyces cerevisiae alpha -Factor Receptor (Ste2p), a 7-Transmembrane-segment G Protein-coupled Receptor.
N. E. David, M. Gee, B. Andersen, F. Naider, J. Thorner, and R. C. Stevens (1997)
J. Biol. Chem. 272, 15553-15561
   Abstract »    Full Text »    PDF »
RGS Proteins and Signaling by Heterotrimeric G Proteins.
H. G. Dohlman and J. Thorner (1997)
J. Biol. Chem. 272, 3871-3874
   Full Text »    PDF »
Recombinant Microorganisms as Tools for High Throughput Screening for Nonantibiotic Compounds.
R. D. Klein and T. G. Geary (1997)
J Biomol Screen 2, 41-49
   Abstract »    PDF »
Tethered Ligand Library for Discovery of Peptide Agonists.
J. Chen, H. S. Bernstein, M. Chen, L. Wang, M. Ishii, C. W. Turck, and S. R. Coughlin (1995)
J. Biol. Chem. 270, 23398-23401
   Abstract »    Full Text »    PDF »
Crystal structure of a conserved protease that binds DNA: the bleomycin hydrolase, Gal6.
L Joshua-Tor, H. Xu, S. Johnston, and D. Rees (1995)
Science 269, 945-950
   Abstract »    PDF »
Diversity of G proteins in signal transduction.
M. Simon, M. Strathmann, and N Gautam (1991)
Science 252, 802-808
   Abstract »    PDF »
Activation of Heterotrimeric G-protein Signaling by a Ras-related Protein. IMPLICATIONS FOR SIGNAL INTEGRATION.
M. J. Cismowski, C. Ma, C. Ribas, X. Xie, M. Spruyt, J. S. Lizano, S. M. Lanier, and E. Duzic (2000)
J. Biol. Chem. 275, 23421-23424
   Abstract »    Full Text »    PDF »
Single Amino Acid Substitutions and Deletions That Alter the G Protein Coupling Properties of the V2 Vasopressin Receptor Identified in Yeast by Receptor Random Mutagenesis.
I. Erlenbach, E. Kostenis, C. Schmidt, C. S.-L. Gal, D. Raufaste, M. E. Dumont, M. H. Pausch, and J. Wess (2001)
J. Biol. Chem. 276, 29382-29392
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)