Control of yeast mating signal transduction by a mammalian beta 2-adrenergic receptor and Gs alpha subunit.
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.
King
K
K
Department of Medicine (Cardiology), Howard Hughes Medical Institute, Duke University Medical Center, Durham, NC 27710.
Dohlman
H G
HG
Thorner
J
J
Caron
M G
MG
Lefkowitz
R J
RJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D002462:Cell Membrane / Q000502:physiology; D019204:GTP-Binding Proteins / Q000235:genetics / Q000502:physiology*; D015870:Gene Expression; D006801:Humans; D020120:Iodocyanopindolol; D007700:Kinetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D010869:Pindolol / Q000031:analogs & derivatives / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011943:Receptors, Adrenergic, beta / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology*; D015398:Signal Transduction
D046911:Macromolecular Substances; D011943:Receptors, Adrenergic, beta; D011993:Recombinant Fusion Proteins; D020120:Iodocyanopindolol; D010869:Pindolol; D019204:GTP-Binding Proteins