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PMID: 1464310 Published · ppublish English Journal Article

Dominant-negative mutants of a yeast G-protein beta subunit identify two functional regions involved in pheromone signalling.

The EMBO journal ·Vol. 11 ·No. 13 ·1992-12-00 ·Pages 4805-13

Leberer E, Dignard D, Hougan L, Thomas DY, Whiteway M

Abstract

The STE4 gene, which encodes the beta subunit of the mating response G-protein in the yeast Saccharomyces cerevisiae, was subjected to a saturation mutagenesis using 'doped' oligodeoxynucleotides. We employed a genetic screen to select dominant-negative STE4 mutants, which when overexpressed from the GAL1 promoter, interfered with the signalling function of the wild type protein. The identified inhibitory amino acid alterations define two small regions that are crucially involved in transmitting the mating signal from G beta to downstream components of the signalling pathway. These results underline the positive signalling role of yeast G beta and assign for the first time the positive signalling function of a G-protein beta subunit to specific structural features.

Related Genes
GAL1 STE18 STE4
MeSH Terms
Amino Acid Sequence GTP-Binding Protein beta Subunits GTP-Binding Proteins/genetics,metabolism Genes, Dominant Heterotrimeric GTP-Binding Proteins Molecular Sequence Data Mutation Pheromones/metabolism Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins
Chemicals
GTP-Binding Protein beta Subunits Pheromones Saccharomyces cerevisiae Proteins Ste4 protein, S cerevisiae GTP-Binding Proteins Heterotrimeric GTP-Binding Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Leberer E
Eukaryotic Genetics Group, National Research Council of Canada, Montreal, Quebec.
Dignard D
Hougan L
Thomas D Y
Whiteway M
References (43)
43 references, click to expand
  1. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  2. Common signal transduction system shared by STE2 and STE3 in haploid cells of Saccharomyces cerevisiae: autocrine cell-cycle arrest results from forced expression of STE2.
    EMBO J. 1987 Jan;6(1):249-54 PMID: 15981334
  3. Functional complementation of yeast ste6 by a mammalian multidrug resistance mdr gene.
    Science. 1992 Apr 10;256(5054):232-4 PMID: 1348873
  4. The protein kinase homologue Ste20p is required to link the yeast pheromone response G-protein beta gamma subunits to downstream signalling components.
    EMBO J. 1992 Dec;11(13):4815-24 PMID: 1464311
  5. Signal transduction during pheromone response in yeast.
    Annu Rev Cell Biol. 1991;7:699-728 PMID: 1667085
  6. Activation by G protein beta gamma subunits of agonist- or light-dependent phosphorylation of muscarinic acetylcholine receptors and rhodopsin.
    J Biol Chem. 1992 Feb 5;267(4):2222-7 PMID: 1733928
  7. Sequence of the beta-subunit of the phosphatidylinositol-specific phospholipase C-directed GTP-binding protein from squid (Loligo forbesi) photoreceptors.
    Biochem J. 1991 Jan 1;273(Pt 1):225-8 PMID: 1899186
  8. Diversity of G proteins in signal transduction.
    Science. 1991 May 10;252(5007):802-8 PMID: 1902986
  9. The CDC20 gene product of Saccharomyces cerevisiae, a beta-transducin homolog, is required for a subset of microtubule-dependent cellular processes.
    Mol Cell Biol. 1991 Nov;11(11):5592-602 PMID: 1922065
  10. Overexpression of the STE4 gene leads to mating response in haploid Saccharomyces cerevisiae.
    Mol Cell Biol. 1990 Jan;10(1):217-22 PMID: 2104659
  11. Stoichiometry of G protein subunits affects the Saccharomyces cerevisiae mating pheromone signal transduction pathway.
    Mol Cell Biol. 1990 Feb;10(2):510-7 PMID: 2105453
  12. Regulation of the yeast pheromone response pathway by G protein subunits.
    EMBO J. 1990 Mar;9(3):691-6 PMID: 2107073
  13. Molecular cloning of beta 3 subunit, a third form of the G protein beta-subunit polypeptide.
    Proc Natl Acad Sci U S A. 1990 Mar;87(6):2329-33 PMID: 2107550
  14. Characterization of a G-protein beta-subunit gene from the nematode Caenorhabditis elegans.
    J Mol Biol. 1990 May 5;213(1):17-26 PMID: 2110981
  15. Sites of interaction in the complex between beta- and gamma-subunits of transducin.
    J Biol Chem. 1990 Aug 5;265(22):12995-9 PMID: 2115886
  16. A Chlamydomonas gene encodes a G protein beta subunit-like polypeptide.
    Mol Gen Genet. 1990 May;221(3):443-52 PMID: 2116589
  17. The GTPase superfamily: a conserved switch for diverse cell functions.
    Nature. 1990 Nov 8;348(6297):125-32 PMID: 2122258
  18. Identification of a gene necessary for cell cycle arrest by a negative growth factor of yeast: FAR1 is an inhibitor of a G1 cyclin, CLN2.
    Cell. 1990 Nov 30;63(5):999-1011 PMID: 2147873
  19. Functional inactivation of genes by dominant negative mutations.
    Nature. 1987 Sep 17-23;329(6136):219-22 PMID: 2442619
  20. The product of the PRP4 gene of S. cerevisiae shows homology to beta subunits of G proteins.
    Cell. 1989 Sep 8;58(5):811-2 PMID: 2528410
  21. The STE4 and STE18 genes of yeast encode potential beta and gamma subunits of the mating factor receptor-coupled G protein.
    Cell. 1989 Feb 10;56(3):467-77 PMID: 2536595
  22. Constitutive mutants in the yeast pheromone response: ordered function of the gene products.
    Cell. 1989 Feb 10;56(3):479-86 PMID: 2644047
  23. G-protein subunits. Who carries what message?
    Nature. 1989 Feb 9;337(6207):504 PMID: 2644543
  24. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  25. Pheromonal regulation and sequence of the Saccharomyces cerevisiae SST2 gene: a model for desensitization to pheromone.
    Mol Cell Biol. 1987 Dec;7(12):4169-77 PMID: 2830483
  26. Yeast peptide pheromones, a-factor and alpha-factor, activate a common response mechanism in their target cells.
    Cell. 1986 Dec 26;47(6):929-37 PMID: 3022943
  27. Transcriptional regulation of an hsp70 heat shock gene in the yeast Saccharomyces cerevisiae.
    Mol Cell Biol. 1987 May;7(5):1906-16 PMID: 3037338
  28. Repetitive segmental structure of the transducin beta subunit: homology with the CDC4 gene and identification of related mRNAs.
    Proc Natl Acad Sci U S A. 1986 Apr;83(7):2162-6 PMID: 3083416
  29. Beta-subunits of the human liver Gs/Gi signal-transducing proteins and those of bovine retinal rod cell transducin are identical.
    FEBS Lett. 1986 Oct 27;207(2):187-92 PMID: 3095147
  30. Distinct forms of the beta subunit of GTP-binding regulatory proteins identified by molecular cloning.
    Proc Natl Acad Sci U S A. 1987 Jun;84(11):3792-6 PMID: 3108879
  31. The yeast SCG1 gene: a G alpha-like protein implicated in the a- and alpha-factor response pathway.
    Cell. 1987 Sep 25;50(7):1001-10 PMID: 3113738
  32. GPA1, a haploid-specific essential gene, encodes a yeast homolog of mammalian G protein which may be involved in mating factor signal transduction.
    Cell. 1987 Sep 25;50(7):1011-9 PMID: 3113739
  33. The Saccharomyces cerevisiae BAR1 gene encodes an exported protein with homology to pepsin.
    Proc Natl Acad Sci U S A. 1988 Jan;85(1):55-9 PMID: 3124102
  34. Cloning of a Drosophila melanogaster guanine nucleotide regulatory protein beta-subunit gene and characterization of its expression during development.
    Proc Natl Acad Sci U S A. 1988 Oct;85(19):7134-8 PMID: 3140235
  35. A simple and efficient procedure for generating random point mutations and for codon replacements using mixed oligodeoxynucleotides.
    DNA. 1988 Mar;7(2):127-34 PMID: 3282853
  36. Two genes required for cell fusion during yeast conjugation: evidence for a pheromone-induced surface protein.
    Mol Cell Biol. 1987 Jul;7(7):2316-28 PMID: 3302672
  37. Rapid and efficient site-specific mutagenesis without phenotypic selection.
    Methods Enzymol. 1987;154:367-82 PMID: 3323813
  38. One-step gene disruption in yeast.
    Methods Enzymol. 1983;101:202-11 PMID: 6310324
  39. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  40. Comparison of dose-response curves for alpha factor-induced cell division arrest, agglutination, and projection formation of yeast cells. Implication for the mechanism of alpha factor action.
    J Biol Chem. 1983 Nov 25;258(22):13849-56 PMID: 6358212
  41. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance.
    Mol Gen Genet. 1984;197(2):345-6 PMID: 6394957
  42. Use of unpurified synthetic deoxynucleotide primers for rapid dideoxynucleotide chain termination sequencing.
    DNA. 1984 Aug;3(4):339-43 PMID: 6489095
  43. Hormonal stimulation of adenylyl cyclase through Gi-protein beta gamma subunits.
    Nature. 1992 Mar 12;356(6365):159-61 PMID: 1312225
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1992-12-00
Pages
4805-13
Language
English
Region
England
NLM ID
8208664
PMCID
PMC556956
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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