• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. May 17, 1999; 18(10): 2756–2763.
PMCID: PMC1171357

A constitutively active G-protein-coupled receptor causes mating self-compatibility in the mushroom Coprinus.

Abstract

In the mushroom Coprinus cinereus, the multiallelic B mating type genes are predicted to encode a large family of seven-transmembrane domain receptors and CaaX-modified pheromones. We have shown that a single amino acid change Q229P in transmembrane domain VI of one receptor confers a self-compatible mating phenotype. Using a heterologous yeast assay, we have demonstrated that this C.cinereus pheromone receptor is a G-protein-coupled receptor and that the Q229P mutation is constitutively activating. A C.cinereus pheromone precursor was processed to an active species specifically in yeast MATa cells and activated the co-expressed wild-type receptor. Yeast cells expressing the wild-type receptor were used to test the activity of synthetic peptides, enabling us to predict the structure of the mature C.cinereus pheromone and to show that the Q229P mutation does not compromise normal receptor function.

Full Text

The Full Text of this article is available as a PDF (337K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bölker M, Kahmann R. Sexual pheromones and mating responses in fungi. Plant Cell. 1993 Oct;5(10):1461–1469. [PMC free article] [PubMed]
  • Bölker M, Urban M, Kahmann R. The a mating type locus of U. maydis specifies cell signaling components. Cell. 1992 Feb 7;68(3):441–450. [PubMed]
  • Bourne HR. How receptors talk to trimeric G proteins. Curr Opin Cell Biol. 1997 Apr;9(2):134–142. [PubMed]
  • Caldwell GA, Wang SH, Xue CB, Jiang Y, Lu HF, Naider F, Becker JM. Molecular determinants of bioactivity of the Saccharomyces cerevisiae lipopeptide mating pheromone. J Biol Chem. 1994 Aug 5;269(31):19817–19825. [PubMed]
  • Caldwell GA, Naider F, Becker JM. Fungal lipopeptide mating pheromones: a model system for the study of protein prenylation. Microbiol Rev. 1995 Sep;59(3):406–422. [PMC free article] [PubMed]
  • Casselton LA, Olesnicky NS. Molecular genetics of mating recognition in basidiomycete fungi. Microbiol Mol Biol Rev. 1998 Mar;62(1):55–70. [PMC free article] [PubMed]
  • Clare JJ, Romanos MA, Rayment FB, Rowedder JE, Smith MA, Payne MM, Sreekrishna K, Henwood CA. Production of mouse epidermal growth factor in yeast: high-level secretion using Pichia pastoris strains containing multiple gene copies. Gene. 1991 Sep 15;105(2):205–212. [PubMed]
  • Conklin BR, Bourne HR. Structural elements of G alpha subunits that interact with G beta gamma, receptors, and effectors. Cell. 1993 May 21;73(4):631–641. [PubMed]
  • Dowell SJ, Bishop AL, Dyos SL, Brown AJ, Whiteway MS. Mapping of a yeast G protein betagamma signaling interaction. Genetics. 1998 Dec;150(4):1407–1417. [PMC free article] [PubMed]
  • Dube P, Konopka JB. Identification of a polar region in transmembrane domain 6 that regulates the function of the G protein-coupled alpha-factor receptor. Mol Cell Biol. 1998 Dec;18(12):7205–7215. [PMC free article] [PubMed]
  • Hagen DC, McCaffrey G, Sprague GF., Jr Evidence the yeast STE3 gene encodes a receptor for the peptide pheromone a factor: gene sequence and implications for the structure of the presumed receptor. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1418–1422. [PMC free article] [PubMed]
  • Herskowitz I. MAP kinase pathways in yeast: for mating and more. Cell. 1995 Jan 27;80(2):187–197. [PubMed]
  • Jensen R, Sprague GF, Jr, Herskowitz I. Regulation of yeast mating-type interconversion: feedback control of HO gene expression by the mating-type locus. Proc Natl Acad Sci U S A. 1983 May;80(10):3035–3039. [PMC free article] [PubMed]
  • Kang YS, Kane J, Kurjan J, Stadel JM, Tipper DJ. Effects of expression of mammalian G alpha and hybrid mammalian-yeast G alpha proteins on the yeast pheromone response signal transduction pathway. Mol Cell Biol. 1990 Jun;10(6):2582–2590. [PMC free article] [PubMed]
  • King K, Dohlman HG, Thorner J, Caron MG, Lefkowitz RJ. Control of yeast mating signal transduction by a mammalian beta 2-adrenergic receptor and Gs alpha subunit. Science. 1990 Oct 5;250(4977):121–123. [PubMed]
  • Konopka JB, Margarit SM, Dube P. Mutation of Pro-258 in transmembrane domain 6 constitutively activates the G protein-coupled alpha-factor receptor. Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6764–6769. [PMC free article] [PubMed]
  • Krüger J, Loubradou G, Regenfelder E, Hartmann A, Kahmann R. Crosstalk between cAMP and pheromone signalling pathways in Ustilago maydis. Mol Gen Genet. 1998 Nov;260(2-3):193–198. [PubMed]
  • Kurjan J. The pheromone response pathway in Saccharomyces cerevisiae. Annu Rev Genet. 1993;27:147–179. [PubMed]
  • Leberer E, Thomas DY, Whiteway M. Pheromone signalling and polarized morphogenesis in yeast. Curr Opin Genet Dev. 1997 Feb;7(1):59–66. [PubMed]
  • Michaelis S, Herskowitz I. The a-factor pheromone of Saccharomyces cerevisiae is essential for mating. Mol Cell Biol. 1988 Mar;8(3):1309–1318. [PMC free article] [PubMed]
  • Milano CA, Allen LF, Rockman HA, Dolber PC, McMinn TR, Chien KR, Johnson TD, Bond RA, Lefkowitz RJ. Enhanced myocardial function in transgenic mice overexpressing the beta 2-adrenergic receptor. Science. 1994 Apr 22;264(5158):582–586. [PubMed]
  • Milligan G, Marshall F, Rees S. G16 as a universal G protein adapter: implications for agonist screening strategies. Trends Pharmacol Sci. 1996 Jul;17(7):235–237. [PubMed]
  • Moore TD, Edman JC. The alpha-mating type locus of Cryptococcus neoformans contains a peptide pheromone gene. Mol Cell Biol. 1993 Mar;13(3):1962–1970. [PMC free article] [PubMed]
  • Mumberg D, Müller R, Funk M. Yeast vectors for the controlled expression of heterologous proteins in different genetic backgrounds. Gene. 1995 Apr 14;156(1):119–122. [PubMed]
  • Nomoto S, Nakayama N, Arai K, Matsumoto K. Regulation of the yeast pheromone response pathway by G protein subunits. EMBO J. 1990 Mar;9(3):691–696. [PMC free article] [PubMed]
  • Offermanns S, Simon MI. G alpha 15 and G alpha 16 couple a wide variety of receptors to phospholipase C. J Biol Chem. 1995 Jun 23;270(25):15175–15180. [PubMed]
  • O'Shea SF, Chaure PT, Halsall JR, Olesnicky NS, Leibbrandt A, Connerton IF, Casselton LA. A large pheromone and receptor gene complex determines multiple B mating type specificities in Coprinus cinereus. Genetics. 1998 Mar;148(3):1081–1090. [PMC free article] [PubMed]
  • Price LA, Kajkowski EM, Hadcock JR, Ozenberger BA, Pausch MH. Functional coupling of a mammalian somatostatin receptor to the yeast pheromone response pathway. Mol Cell Biol. 1995 Nov;15(11):6188–6195. [PMC free article] [PubMed]
  • Regenfelder E, Spellig T, Hartmann A, Lauenstein S, Bölker M, Kahmann R. G proteins in Ustilago maydis: transmission of multiple signals? EMBO J. 1997 Apr 15;16(8):1934–1942. [PMC free article] [PubMed]
  • Rothstein R. Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast. Methods Enzymol. 1991;194:281–301. [PubMed]
  • Scheer A, Cotecchia S. Constitutively active G protein-coupled receptors: potential mechanisms of receptor activation. J Recept Signal Transduct Res. 1997 Jan-May;17(1-3):57–73. [PubMed]
  • Sikorski RS, Hieter P. 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. [PMC free article] [PubMed]
  • Spalding TA, Burstein ES, Brauner-Osborne H, Hill-Eubanks D, Brann MR. Pharmacology of a constitutively active muscarinic receptor generated by random mutagenesis. J Pharmacol Exp Ther. 1995 Dec;275(3):1274–1279. [PubMed]
  • Spalding TA, Burstein ES, Wells JW, Brann MR. Constitutive activation of the m5 muscarinic receptor by a series of mutations at the extracellular end of transmembrane 6. Biochemistry. 1997 Aug 19;36(33):10109–10116. [PubMed]
  • Spellig T, Bölker M, Lottspeich F, Frank RW, Kahmann R. Pheromones trigger filamentous growth in Ustilago maydis. EMBO J. 1994 Apr 1;13(7):1620–1627. [PMC free article] [PubMed]
  • Tam A, Nouvet FJ, Fujimura-Kamada K, Slunt H, Sisodia SS, Michaelis S. Dual roles for Ste24p in yeast a-factor maturation: NH2-terminal proteolysis and COOH-terminal CAAX processing. J Cell Biol. 1998 Aug 10;142(3):635–649. [PMC free article] [PubMed]
  • Tseng CC, Lin L. A point mutation in the glucose-dependent insulinotropic peptide receptor confers constitutive activity. Biochem Biophys Res Commun. 1997 Mar 6;232(1):96–100. [PubMed]
  • Vaillancourt LJ, Raper CA. Pheromones and pheromone receptors as mating-type determinants in basidiomycetes. Genet Eng (N Y) 1996;18:219–247. [PubMed]
  • Vaillancourt LJ, Raudaskoski M, Specht CA, Raper CA. Multiple genes encoding pheromones and a pheromone receptor define the B beta 1 mating-type specificity in Schizophyllum commune. Genetics. 1997 Jun;146(2):541–551. [PMC free article] [PubMed]
  • Wach A. PCR-synthesis of marker cassettes with long flanking homology regions for gene disruptions in S. cerevisiae. Yeast. 1996 Mar 15;12(3):259–265. [PubMed]
  • Wendland J, Vaillancourt LJ, Hegner J, Lengeler KB, Laddison KJ, Specht CA, Raper CA, Kothe E. The mating-type locus B alpha 1 of Schizophyllum commune contains a pheromone receptor gene and putative pheromone genes. EMBO J. 1995 Nov 1;14(21):5271–5278. [PMC free article] [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...