Skip to main content
SpringerLink
Log in

A ste12 allele having a differential effect on a versus α cells

  • Original Paper
  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Abstract

The transcriptional activator Ste12p is a key component of the yeast pheromone response pathway: phosphorylated as a consequence of signal transduction, it activates transcription of genes that promote mating and the subsequent fusion of the two cell types a and α. Activation by Ste12p requires three types of protein-protein interaction between DNA-binding activator proteins: (1) Ste12p by itself can induce non-cell-type-specific genes involved in mating; (2) cooperation of the transactivator Mcm1p with Ste12p induces a-specific genes; and (3) formation of a complex of the activator proteins Mcm1p and α1 (a transcriptional activator of α-specific genes) with Ste12p is believed to induce α-specific genes. We isolated and characterized a partially functional ste12 allele (ste12-T50), that is defective only in the activation of α-specific genes. ste12-T50 was isolated as a second-site mutation conferring the a mating phenotype on matα2 mutant cells. In matα2 cells, where due to the lack of repressor, α2, both sets of cell-type-specific genes are expressed, ste12-T50 apparently tips the balance in favor of a-specific gene expression. Thus, matα2 ste12-T50 cells mate like a cells. Additional ste12 mutants that confer the a mating phenotype on matα2 cells have also been isolated.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bender A, Sprague GF Jr (1986) Yeast peptide pheromones, a-factor and α-factor, activate a common response mechanism in their target cells. Cell 47:929–937

    Google Scholar 

  • Bender A, Sprague GF Jr (1987) MATα1 protein, a yeast transcription activator, binds synergistically with a second protein to a set of cell-type-specific genes. Cell 50:681–691

    Google Scholar 

  • Bender A, Sprague GF Jr (1989) Pheromones and pheromone receptors are the primary determinants of mating specificity in the yeast Saccharomyces cerevisiae. Genetics 121:463–476

    Google Scholar 

  • Boeke JD, LaCroute F, Fink GR (1984) A positive selection for mutants lacking orotidine-5′-phosphate decarboxylase activity in yeast: 5-fluoroorotic acid resistance. Mol Gen Genet 197: 345–346

    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 14:8463–8475

    Google Scholar 

  • Cairns BR, Ramer SW, Kornberg RD (1992) Order of action of components in the yeast pheromone response pathway revealed with a dominant allele of the STE11 kinase and the multiple phosphorylation of the STE7 kinase. Genes Dev 6:1305–1318

    Google Scholar 

  • Cross FR (1990) The DAF2–2 mutation, a dominant inhibitor of the STE4 step in the α-factor signaling pathway of Saccharomyces cerevisiae MATα cells. Genetics 126: 301–308

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Dolan JW, Kirkman C, Fields S (1989) The yeast STE12 protein binds to the DNA sequence mediating pheromone induction. Proc Natl Acad Sci USA 86:5703–5707

    Google Scholar 

  • Errede B, Ammerer G (1989) STE12, a protein involved in cell-type-specific transcription and signal transduction in yeast, is part of protein-DNA complexes. Genes Dev 3:1349–1361

    Google Scholar 

  • Fields S, Herskowitz I (1987) Regulation by the yeast mating-type locus of STE12, a gene required for cell-type-specific expression. Mol Cell Biol 7:3818–3821

    Google Scholar 

  • Hagen DC, McCaffrey G, Sprague GF Jr (1991) Pheromone response elements are necessary and sufficient for basal and pheromone-induced transcription of the FUS1. Mol Cell Biol 11:2952–2961

    Google Scholar 

  • Hartwell LH (1980) Mutants of S. cerevisiae unresponsibe to cell division control by polypeptide mating pheromone. J Cell Biol 85:811–823

    Google Scholar 

  • Hwang-Shum J-J, Hagen DC, Jarvis EE, Westby CA, Sprague GF Jr (1991) Relative contributions of MCM1 and STE12 to transcriptional activation of a- and α-specific genes from Saccharomyces cerevisiae. Mol Gen Genet 227:197–204

    Google Scholar 

  • Jarvis EE, Hagen DC, Sprague GF Jr (1988) Identification of a DNA segment that is necessary and sufficient for α-specific gene control in Saccharomyces cerevisiae: implications for regulation of α-specific and a-specific genes. Mol Cell Biol 8:309–320

    Google Scholar 

  • Jenness DD, Goldman BS, Hartwell LH (1987) Saccharomyces cerevisiae mutants unresponsive to α-factor pheromone: α-factor binding and extragenic suppression. Mol Cell Biol 7:1311–1319

    Google Scholar 

  • Kronstad JW, Holly JA, MacKay VL (1987) A yeast operator overlaps an upstream activation site. Cell 50: 369–377

    Google Scholar 

  • Lawrence CW (1991) Classical mutagenesis techniques. Methods Enzymol 194:273–281

    Google Scholar 

  • Leberer E, Dignard D, Harcus D, Thomas DY, Whiteway M (1992) The protein kinase homologue Ste20p is required to link the yeast pheromone response G-protein βγ subunits to downstream signalling components. EMBO J 11: 4815–4824

    Google Scholar 

  • MacKay V, Manney T (1974a) Mutations affecting sexual conjugation and related processes in Saccharomyces cerevisiae. I. Isolation and phenotypic characterization of nonmating mutants. Genetics 76:255–271

    Google Scholar 

  • MacKay V, Manney TR (1974b) Mutations affecting sexual conjugation and related processes in Saccharomyces cerevisiae. II. Genetic analysis of nonmating mutants. Genetics 76:273–288

    Google Scholar 

  • Marsh L, Neiman AM, Herskowitz I (1991) Signal transduction during pheromone response in yeast. Annu Rev Cell Biol 7: 699–728

    Google Scholar 

  • Miyajima I, Nakafuku M, Nakayama N, Brenner C, Miyajima A, Kaibuchi K, Arai K-i, Kaziro Y (1987) GPA1, a haploid-specific 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 

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

    Google Scholar 

  • Nakayama N, Miyajima A, Arai K (1987) 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 6:249–254

    Google Scholar 

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

    Google Scholar 

  • Oliver SG (1992) The complete DNA sequence of yeast chromosome III. Nature 357:38–46

    Google Scholar 

  • Passmore S, Elble R, Tye B-K (1989) A protein involved in minichromosome maintenance in yeast binds a transcriptional enhancer conserved in eukaryotes. Genes Dev 3:921–935

    Google Scholar 

  • Rhodes N, Connell L, Errede B (1990) STE11 is a protein kinase required for cell-type-specific transcription and signal transduction in yeast. Genes Dev 4:1862–1874

    Google Scholar 

  • Rose M, Novick P, Thomas JH, Botstein D, Fink G (1987) A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector. Gene 60:237–243

    Google Scholar 

  • Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101: 202–211

    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 

  • Schmitt ME, Brown TA, Trumpower BL (1990) A rapid and simple method for preparation of RNA from Saccharomyces cerevisiae. Nucleic Acids Res 18:3091

    Google Scholar 

  • Sengupta P, Cochran BH (1990) The PRE and PQ Box are functionally distinct yeast pheromone response elements. Mol Cell Biol 10:6809–6812

    Google Scholar 

  • Sengupta P, Cochran BH (1991) MATα1 can mediate gene activation by a-mating factor. Genes Dev 5:1924–1934

    Google Scholar 

  • Sikorski RS, Boeko JD (1991) In vitro mutagenesis and plasmid shuffling: from cloned gene to mutant yeast. Methods Enzymol 194:302–318

    Google Scholar 

  • Sikorski RS, Hieter P (1989) A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122:19–27

    Google Scholar 

  • Song O-k, Dolan JW, Yuan Y-l, Fields S (1991) Pheromone-dependent phosphorylation of the yeast STE12 protein correlates with transcriptional activation. Genes Dev 5: 741–750

    Google Scholar 

  • Sprague GF (1990) Combinatorial associations of regulatory proteins and the control of cell type in yeast. Adv Genet 27: 33–62

    Google Scholar 

  • Sprague GF Jr, Herskowitz I (1981) Control of yeast cell type by the mating type focus. I. Identification and control of expression of the a-specific gene BAR1. J Mol Biol 153:305–321

    Google Scholar 

  • Strathern JN, Hicks J, Herskowitz I (1981) Control of cell-type in yeast by the mating type locus. The α1–α2 hypothesis. J Mol Biol 147:357–372

    Google Scholar 

  • Tan S, Richmond TJ (1990) DNA binding-induced conformational change of the yeast transcriptional activator PRTF. Cell 62: 367–377

    Google Scholar 

  • Tan S, Ammerer G, Richmond TJ (1988) Interactions of purified transcription factors: binding of yeast MATα1 and PRTF to cell type-specific, upstream activating sequences. EMBO J 7: 4255–4264

    Google Scholar 

  • Teague MA, Chaleff DT, Errede B (1986) Nucleotide sequence of the yeast regulatory gene STE7 predicts a protein homologous to protein kinases. Proc Natl Acad Sci USA 83:7371–7375

    Google Scholar 

  • Van Arsdell SW, Thorner J (1987) Hormonal regulation of gene expression in yeast. In: Granner D, Rosenfeld MG, Chang S (eds) Transcriptional control mechanisms. Liss, New York, pp 325–332

    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 

  • Yuan Y-L, Fields S (1991) Properties of the DNA-binding domain of the Saccharomyces cerevisiae STE12 protein. Mol Cell Biol 11:5910–5918

    Google Scholar 

  • Yuan Y-L, Stoke IL, Fields S (1983) Coupling of cell-identity to signal response in yeast: interaction between the α1 and STE12 proteins. Genes Dev 7:1584–1597

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratory of Eukaryotic Gene Expression, NCI-Frederick Cancer Research and Development Center, ABL-Basic Research Program, P.O. Box B, 21702-1201, Frederick, Maryland, USA

    Salome D. La Roche, Brenda K. Shafer & Jeffrey N. Strathern

Authors
  1. Salome D. La Roche
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Brenda K. Shafer
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jeffrey N. Strathern
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by C. P. Hollenberg

Rights and permissions

Reprints and permissions

About this article

Cite this article

La Roche, S.D., Shafer, B.K. & Strathern, J.N. A ste12 allele having a differential effect on a versus α cells. Molec. Gen. Genet. 246, 80–90 (1995). https://doi.org/10.1007/BF00290136

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00290136

Key words

Search

Navigation