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STE50, a novel gene required for activation of conjugation at an early step in mating in Saccharomyces cerevisiae

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Summary

A new gene, STE50, which plays an essential role in cell differentiation in Saccharomyces cerevisiae was detected and analysed. STE50 expression is not cell type-specific and its expression in MAT a and MATα cells is unaffected by pheromones. When present on a high copy number plasmid, STE50 causes supersensitivity to α-pheromone, and increases the level of α-pheromone-induced transcription of FUS1 in haploid a cells. Mutants bearing either of the two gene disruptions, ste50-1 or ste50-2, are sterile and have a modulated sensitivity to α-pheromone. The overexpression of STE4 (Gβ) in wild-type cells elicits a constitutive growth arrest signal, however this phenotype is suppressed by a C-terminal truncation mutation in STE50 (ste50-2). In contrast, the constitutive activation of the pheromone response pathway caused by disruption of GPA1 (Gα) is not suppressed in ste50-2 mutants. The ste50-2 mutation partially suppresses the desensitisation defect of the sst2-1 mutation, and the resulting ste50-2 sst2-1 mutants restore fertility. Our result sindicate that the ste50-2 mutant may have a defect in adaptation (hyperadaptation), and suggest a possible interaction of STE50-2 with the Gα subunit of the G protein.

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References

  • Blinder D, Bouvier S, Jenness DD (1989) Constitutive mutants in the yeast pheromone response: ordered function of the gene products. Cell 56:479–486

    Google Scholar 

  • Blumer KJ, Thorner J (1990) β and γ subunits of a yeast guanine nucleotide-binding protein are not essential for membrane association of the α subunit but are required for receptor coupling. Proc Natl Acad Sci USA 87:4363–4367

    Google Scholar 

  • Burkholder AC, Hartwell LH (1985) The yeast α-factor receptor: Structural properties deduced from the sequence of the STE2 gene. Nucleic Acids Res 13:8463–8475

    Google Scholar 

  • Chan RK, Otte CA (1982) Isolation and genetic analysis of Saccharomyces cerevisiae mutants supersensitive to G1 arrest by a-factor and α-factor pheromones. Mol Cell Biol 2:11–20

    Google Scholar 

  • Cole GM, Stone DE, Reed SI (1990) Stoichiommetry of G protein subunit affects the Saccharomyces cerevisiae mating pheromone signal transduction pathway. Mol Cell Biol 10:510–5417

    Google Scholar 

  • Cross F, Hartwell LH, Jackson C, Konopka JB (1988) Conjugation in Saccharomyces cerevisiae. Annu Rev Cell Biol 4:429–457

    Google Scholar 

  • Dietzel C, Kurjan J (1987) The yeast SCG1 gene: a Gα-like protein implicated in the a- and α-factor response pathway. Cell 50:1001–1010

    Google Scholar 

  • Dohmen RJ, Strasser AWM, Honer CB, Hollenberg CP (1991) An efficient transformation procedure enabling long term storage of competent cells of various yeast genera. Yeast 7:691–692

    Google Scholar 

  • Elion EA, Grisafi PL, Fink GR (1990) FUS3 encodes a cdc2/CDC28-related kinase required for the transition from mitosis into conjugation. Cell 60:649–664

    Google Scholar 

  • Hagen DC, McCaffrey G, Sprague GFJ (1986) Evidence that 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 USA 83:1418–1422

    Google Scholar 

  • Hartwell LH, Culotti J, Pringle JR, Reid BJ (1974) Genetic control of the cell division cycle in yeast. Science 183:46–51

    Google Scholar 

  • Hartwell LH, Weinert TA (1989) Checkpoints: Controls that ensure the order of cell cycle events. Science 246:629–634

    Google Scholar 

  • Herskowitz I (1990) A regulatory hierarchy for cell specialisation in yeast. Nature 342:749–757

    Google Scholar 

  • Hill JE, Myers AM, Koerner TJ, Tzagoloff A (1986) Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast 2:163–167

    Google Scholar 

  • Irie K, Nomoto S, Miyajima I, Matsumoto K (1991) SGV1 encodes a CDC28/cdc2-related kinase required for a Gα subunitmediated adaptive response to pheromone in S. cerevisiae. Cell 65:785–795

    Google Scholar 

  • Jackson CL, Konopka JB, Hartwell LH (1991) S. cerevisiae α-pheromone receptors activate a novel signal transduction pathway for mating partner discrimination. Cell 67:389–402

    Google Scholar 

  • Jahng K-Y, Ferguson J, Reed SI (1988) Mutations in a gene encoding the a subunit of Saccharomyces cerevisiae G protein indicate a role in mating pheromone signalling. Mol Cell Biol 8:2484–2493

    Google Scholar 

  • Klebe RJ, Harriss JV, Sharp ZD, Douglas MG (1983) A general method for polyethylene-glycol-induced genetic transformation of bacteria and yeast. Gene 25:333–341

    Google Scholar 

  • Kurjan J, Dietzel C (1987) Pheromone regulation and sequence of the Saccharomyces cerevisiae SST2 gene: a model for desensitisation to pheromone. Mol Cell Biol 7:4169–4177

    Google Scholar 

  • Kurjan J, Hirsch JP, Dietzel C (1991) Mutations in the guanine nucleotide-binding domains of a yeast Gα protein confer a constitutive or uninducible state to the pheromone response pathway. Genes Dev 5:475–483

    Google Scholar 

  • Michaelis S, Herskowitz I (1988) The α-factor pheromone of Saccharomyces cerevisiae is essential for mating. Mol Cell Biol 8:1309–1318

    Google Scholar 

  • Miyajima I, Arai K-I, Matsumoto K (1989) GPA1 Val-50 mutation in the mating-factor signalling pathway in Saccharomyces cerevisiae. Mol Cell Biol 9:2289–2297

    Google Scholar 

  • Miyajima I, Nakafuku M, Nakayama N, Brenner C, Miyajima A, Kaibuchi K, Arai K, Kaziro Y, Matsumoto K (1987) GPA1, a haploidspecific essential gene, encodes a yeast homolog of mammalian G protein which may be involved in mating factor signal transduction. Cell 50:1011–1019

    Google Scholar 

  • Myers AM, Tzagoloff A, Kinney DM, Lusty CJ (1986) Yeast shuttle and integrative vectors with multiple cloning sites suitable for construction of lacZ fusions. Gene 45:299–310

    Google Scholar 

  • Nakafuku M, Itoh H, Nakamura S, Kaziro Y (1987) Occurrence in Saccharomyces cerevisiae of a gene homologous to the cDNA coding the α subunit of mammalian G protein. Proc Natl Acad Sci USA 84:2140–2144

    Google Scholar 

  • Nakayama N, Kaziro Y, Arai K, Matsumoto K (1988) Role of STE genes in the mating factor signalling pathway mediated by GPA1 in Saccharomyces cerevisiae. Mol Cell Biol 8:3777–3783

    Google Scholar 

  • Nakayama N, Miyajima A, Arai K (1985) Nucleotide sequence of STE2 and STE3, cell type-specific sterile genes from Saccharomyces cerevisiae. EMBO J 4:2643–2648

    Google Scholar 

  • Nomoto S, Nakayama N, Arai K, Matsumoto K (1990) Regulation of the yeast pheromone response pathway by G protein subunits. EMBO J 9:691–696

    Google Scholar 

  • Otte CA, Chan RK (1982) Physiological characterisation of Saccharomyces cerevisiae mutants supersensitive to G1 arrest by a- and α-factor pheromone. Mol Cell Biol 2:21–29

    Google Scholar 

  • Ramezani Rad M, Lutzenkirchen K, Xu G, Kleinhans U, Hollenberg CP (1991) The complete sequence of a 11953 by fragment from CIG on chromosome 111 encompasses four new open reading frames. Yeast 7:533–538

    Google Scholar 

  • Ramezani Rad M, Xu G, Hollenberg CP (1990) Yeast STE50, a gene implicated in the a- and α-factor response pathway. Yeast 6 (special issue):213

    Google Scholar 

  • Reneke JE, Blumer K, Courchesne WE, Thorner J (1988) The carboxy-terminal segment of the yeast α-factor receptor is a regulatory domain. Cell 55:221–234

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Sena EP, Radin DN, Fogel S (1973) Synchronous mating in yeast. Proc Natl Acad Sci USA 70:1373–1377

    Google Scholar 

  • Sherman F, Fink GR, Hicks JB (1986) Laboratory course manual for methods in yeast genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Struhl K (1985) Nucleotide sequence and transcriptional mapping of the yeast pet56-his3-dedl gene region. Nucleic Acids Res 13:8587–8601

    Google Scholar 

  • Tanaka K, Nakafuku M, Satoh T, Marshall MS, Gibbs JB (1991) S. cerevisiae genes IRA1 and IRA2 encode proteins that may be functionally equivalent to mammalian ras GTPase activating protein. Cell 60:803–807

    Google Scholar 

  • Trueheart J, Boeke JD, Fink GR (1987) Two genes required for cell fusion during yeast conjugation: evidence for a pheromoneinduced surface protein. Mol Cell Biol 7:2316–2328

    Google Scholar 

  • Trueheart J, Fink GR (1989) The yeast cell fusion protein FUS1 is O-glycosylated and spans the plasma membrane. Proc Natl Acad Sci USA 86:9916–9920

    Google Scholar 

  • Whiteway M, Hougan L, Dignard D, Thomas DY, Bell L, Saari GC, Grant FJ, O'Hara P, MacKay VL (1989) The STE4 and STE18 genes of yeast encode potential β and γ subunits of the mating factor receptor-coupled G protein. Cell 56:467–477

    Google Scholar 

  • Whiteway M, Hougan L, Thomas DY (1990) Overexpression of the STE4 gene leads to mating response in haploid Saccharomyces cerevisiae. Mol Cell Biol 10:217–222

    Google Scholar 

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Authors and Affiliations

  1. Institut für Mikrobiologie, Heinrich- Heine-Universität, Universitätsstraße 1, W-4000, Düsseldorf 1, Germany

    Massoud Ramezani Rad, Gang Xu & Cornelis P. Hollenberg

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  1. Massoud Ramezani Rad
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  2. Gang Xu
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  3. Cornelis P. Hollenberg
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Communicated by D.Y. Thomas

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Ramezani Rad, M., Xu, G. & Hollenberg, C.P. STE50, a novel gene required for activation of conjugation at an early step in mating in Saccharomyces cerevisiae . Molec. Gen. Genet. 236, 145–154 (1992). https://doi.org/10.1007/BF00279653

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  • DOI: https://doi.org/10.1007/BF00279653

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