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Constitutive activation of the Saccharomyces cerevislae mating response pathway by a MAP kinase kinase from Candida albicans

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Abstract

The HST7 gene of Candida albicans encodes a protein with structural similarity to MAP kinase kinases. Expression of this gene in Saccharomyces cerevisiae complements disruption of the Ste7 MAP kinase kinase required for both mating in haploid cells and pseudohyphal growth in diploids. However, Hst7 expression does not complement loss of either the Pbs2 (Hog4) MAP kinase kinase required for response to high osmolarity, or loss of the Mkk1 and Mkk2 MAP kinase kinases required for proper cell wall biosynthesis. Intriguingly, HST7 acts as a hyperactive allele of STE7; expression of Hst7 activates the mating pathway even in the absence of upstream signaling components including the Ste7 regulator Ste11, elevates the basal level of the pheromone-inducible FUS1 gene, and amplifies the pseudohyphal growth response in diploid cells. Thus Hst7 appears to be at least partially independent of upstream activators or regulators, but selective in its activity on downstream target MAP kinases. Creation of Hst7/Ste7 hybrid proteins revealed that the C-terminal two-thirds of Hst7, which contains the protein kinase domain, is sufficient to confer this partial independence of upstream activators.

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References

  • Banuett F, Herskowitz I (1994) Identification of Fuz7, a Utsilago maydis MEK/MAPKK homolog required for a-locus-dependent and -independent steps in the fungal life cycle. Genes Dev 8:1367–1378

    Google Scholar 

  • Blenis J (1993) Signal transduction via the MAP kinases: proceed at your own RSK. Proc Natl Acad Sci USA 90:5889–5892

    Google Scholar 

  • Boguslawski G, Polazzi JO (1987) Complete nucleotide sequence of a gene conferring polymyxin B resistance on yeast: similarity of the predicted polypeptide to protein kinases. Proc Natl Acad Sci USA 84:5848–5852

    Google Scholar 

  • Boulton TG, Nye SH, Robbins DJ, Ip NY, Radziejewska E, Morganbesser SD, DePinho RA, Panayotatos N, Cobb, MH Yancopoulos GD (1991) ERKs: a family of protein-serine/threonine kinases that are activated and tyrosine phosphorylated in response to insulin and NGF. Cell 65:677–690

    Google Scholar 

  • Brewster JL, Valoir TD, Dwyer ND, Winter E, Gustin MC (1993) An osmosensing signal transduction pathway in yeast. Science 259:1760–1763

    Google Scholar 

  • Buffo J, Herman MA, Sell DR (1984) A characterization of pHregulated dimorphism in Candida albicans. Mycopathologica 85:21–30

    Google Scholar 

  • Chang F, Herskowitz I (1990) Identification of a gene necessary for cell cycle arrest by a negative growth factor of yeast: FAR1 is an inhibitor of a Gl cyclin, CLN2. Cell 63:999–1011

    Google Scholar 

  • Clark KL, Sprague GF (1989) Yeast pheromone response pathway: characterization of a suppressor that restores mating to receptorless mutants. Mol Cell Biol 9:2682–2694

    Google Scholar 

  • Clark KL, Dignard D, Thomas DY, Whiteway M (1993) Interactions among the subunits of the G protein involved in Saccharomyces cerevisiae mating. Mol Cell Biol 13:1–8

    Google Scholar 

  • Crews CM, Alessandrini A, Erickson RL (1992) The primary structure of MEK, a protein kinase that phosphorylates the ERK1 gene product. Science 258:478–480

    Google Scholar 

  • Dent P, Haser W, Haystead TAJ, Vincent LA, Roberts TM, Sturgill TW (1992) Activation of mitogen-activated protein kinase kinase by v-Raf in NIH 3T3 cells and in vitro. Science 257:1404–1407

    Google Scholar 

  • Elion EA, Satterberg B, Kranz JE (1993) FUS3 phosphorylates multiple components of the mating signal transduction cascade: evidence for STE12 and FAR1. Mol Biol Cell 4:495–510

    Google Scholar 

  • Errede B, Levin DE (1993) A conserved kinase cascade for MAP kinase activation in yeast. Curr Opin Cell Biol 5:254–260

    Google Scholar 

  • Errede B, Gartner A, Zhou Z, Nasmyth K, Ammerer G (1993) MAP kinase-related FUS3 from S. cerevisiae is activated by STE7 in vitro. Nature 362:261–264

    Google Scholar 

  • Gartner A, Nasmyth K, Ammerer G (1992) Signal transduction in Saccharomyces cerevisiae requires tyrosine and threonine phosphorylation of FUS3 and KSS1. Genes Dev 6:1280–1292

    Google Scholar 

  • Gimeno CJ, Ljungdahl PO, Styles CA, Fink GR (1992) Unipolar cell divisions in the yeast S. cerevisiae lead to filamentous growth: regulation by starvation and Ras. Cell 68:1077–1090

    Google Scholar 

  • Hanks SK, Quinn AM (1991) Protein kinase catalytic domain sequence database: identification of conserved features of primary structure and classification of family members. Meth Enzymol 200:38–62

    Google Scholar 

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

    Google Scholar 

  • Irie K, Takase M, Lee KS, Levin DE, Araki H, Matsumoto K, Oshima Y (1993) MKK1 and MKK2, which encode Saccharomyces cerevisiae mitogen-activated protein kinase-kinase homologs, function in the pathway mediated by protein kinase C. Mol Cell Biol 13:3076–3083

    Google Scholar 

  • Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations, J Bacteriol 153:163–168

    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 a Saccharomyces cerevisiae G protein indicate a role in mating pheromone signaling. Mol Cell Biol 8:2484–2493

    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 aspecific and a-specific genes. Mol Cell Biol 8:309–320

    Google Scholar 

  • Kosako H, Nishida E, Gotoh Y (1993) cDNA cloning of MAP kinase kinase reveals cascade pathways in yeast to vertebrates. EMBO J 12:787–794

    Google Scholar 

  • Kyriakis JM, App H, Zhang X-F, Banerjie P, Brautigan DL, Rapp UR, Avruch J (1992) Raf-1 activates MAP kinase-kinase. Nature 358:417–421

    Google Scholar 

  • Lange-Carter CA, Pleiman CM, Gardner AM, Blumer KJ, Johnson GL (1993) A divergence in the MAP kinase regulatory network defined by MEK kinase and RAF. Science 260:315–318

    Google Scholar 

  • Leberer E, Dignard D, Harcus D, Thomas DY, Whiteway M (1992a) 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 

  • Leberer E, Dignard D, Hougan L, Thomas DY, Whiteway M (1992b) Dominant-negative mutants of a yeast G-protein β subunit identify two functional regions involved in pheromone signalling. EMBO J 11:4805–4813

    Google Scholar 

  • Lee KL, Buckler HR, Campbell CC (1975) An amino acid liquid synthetic medium for the development of mycelial and yeast forms of Candida albicans. Sabouraudia 13:148–153

    Google Scholar 

  • Lee KS, Levin DE (1992) Dominant mutations in a gene encoding a putative protein kinase (BCK1) bypass the requirement for a Saccharomyces cerevisiae protein kinase C homolog. Mol Cell Biol 12:172–182

    Google Scholar 

  • Lee KS, Irie K, Gotoh Y, Watanabe Y, Arake H, Nishada E, MatsumotoA K, Levin DE (1993) A yeast mitogen-activated protein kinase homolog (Mpklp) mediates signalling by protein kinase C. Mol Cell Biol 13:3067–3075

    Google Scholar 

  • Lindquist S (1981) Regulation of protein synthesis during heat shock. Nature 293:311–314

    Google Scholar 

  • Liu H, Tyles C, Fink GR (1993) Elements of the yeast pheromone response pathway required for filamentous growth of diploids. Science 262:1714–1744

    Google Scholar 

  • Malathi K, Ganesan K, Datta A (1994) Identification of a putative transcription factor in Candida albicans that can complement the mating defect of Saccharomyces cerevisiae ste12 mutants. J Biol Chem 269:22945–22951

    Google Scholar 

  • Marshall CJ (1994) MAP kinase kinase kinase, MAP kinase kinase and MAP kinase. Curr Op Gen Dev 4:82–89

    Google Scholar 

  • Matsuda S, Gotoh Y, Nishida E (1993) Phosphorylation of Xenopus mitogen-activated protein (MAP) kinase kinase by MAP kinase kinase kinase and MAP kinase. J Biol Chem 268:3277–3281

    Google Scholar 

  • Melton DA, Krieg PA, Rebagliati MR, Mariatis T, Zein K, Green MR (1984) Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res 12:7035–7056

    Google Scholar 

  • Nadin-Davis S, Nasim A (1988) A gene which encodes a predicted protein kinase can restore some functions of the ras gene in fission yeast. EMBO J 7:985–993

    Google Scholar 

  • Neiman AM (1993) Conservation and reiteraion of a kinase cascade. Trends Genet 9:390–394

    Google Scholar 

  • Neiman AM, Herskowitz I (1994) Reconstitution of a yeast protein kinase cascade in vitro: Activation of the yeast MEK homologue STE7 by STE11. Proc Natl Acad Sci USA 91:3398–3402

    Google Scholar 

  • Pelech SL, Sanghera JS (1992) Mitogen-activated protein kinases: versatile transducers for cell signaling. Trends Biochem Sci 17:233–238

    Google Scholar 

  • Peter M, Gartner A, Horecka J, Ammerer G, Herskowitz I (1993) FAR1 links the signal transduction pathway to the cell cycle machinery in yeast. Cell 73:747–760

    Google Scholar 

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

    Google Scholar 

  • Sadhu C, Hoekstra D, McEachern MJ, Reed SI, Hicks JB (1992) A G-protein a subunit from asexual Candida albicans functions in the mating signal transduction pathway of Saccharomyces cerevisiae and is regulated by the a1-α2 repressor. Mol Cell Biol 12:1977–1985

    Google Scholar 

  • Saiki RJ, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491

    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 

  • Seger R, Seger D, Lozeman FJ, Ahn NG, Graves LM, Campbell JS, Ericsson L, Harrylock M, Jensen AM, Krebs EG (1992) Human T-cell mitogen-activated protein kinase kinases are related to yeast signal transduction kinases. J Biol Chem 267:25628–25631

    Google Scholar 

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

    Google Scholar 

  • Sikorski RS, Heiter 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 

  • Slutsky B, Buffo J, Soll DR (1985) High frequency switching of colony morphology in Candida albicans. Science 230:666–669

    Google Scholar 

  • Smith DJ, Cooper M, DeTiani M, Losberger C, Payton MA (1992) The Candida albicans PMMI gene encoding phosphomannomutase complements a Saccharomyces cerevisiae sec53-6 mutation. Curr Genet 22:501–503

    Google Scholar 

  • Soll DR (1986) The regulation of cellular differentiation in the dimorphic yeast Candida albicans. BioEssays 5:5–11

    Google Scholar 

  • Stevenson BJ, Rhodes N, Errede B, Sprague GF Jr (1992) Constitutive mutants of the protein kinase STE11 activate the yeast pheromone response pathway in the absence of the G protein. Genes Dev 6:1293–1304

    Google Scholar 

  • Swoboda RK, Bertram G, Hollander D, Greenspan D, Greenspan JS, Gow NAR, Gooday GW, Brown AJP (1993) Glycolytic enzymes of Candida albicans are nonubiquitous immunogens during candiiasis. Infect Immun 61:4263–4271

    Google Scholar 

  • Swoboda RK, Bertram G, Delbruck S, Ernst JF, Gow NAR, Gooday GW, Brown AJP (1994a) Fluctuations in glycolytic mRNA levels during morphogenesis in Candida albicans reflect underlying changes in growth and not a response to cellular dimorphism. Molec Microbiol 13:663–672

    Google Scholar 

  • Swoboda RK, Bertram G, Colthurst DR, Tuite MF, Gow NAR, Gooday GW, Brown AJP (1994b) Regulation of the gene encoding translation elongation factor 3 during growth and morphogenesis in Candida albicans. Microbiology 140:2611–2616

    Google Scholar 

  • Swoboda RK, Broadbent ID, Bertram G, Gooday GW, Gow NAR, Brown AJP (1995) Structure and regulation of the Candida albicans RP10 gene which encodes an immunogenic protein homologous to Saccharomyces cerevisiae ribosomal protein 10. J Bacteriol 177:1239–1246

    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 

  • Tsuda L, Inoue Y, You M-A, Mizuno M, Hata M, Lim YM, Adachi-Yamada T, Ryo H, Matsamune Y, Nishida Y (1993) A protein kinase similar to MAP kinase activator acts downstream of the Raf kinase in Drosophila. Cell 72:407–414

    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 

  • Whiteway M, Dignard D, Thomas DY (1992) Dominant negative selection of heterologous genes: Isolation of Candida albicans genes that interfere with Saccharomyces cerevisiae mating factorinduced cell cycle arrest. Proc Natl Acad Sci USA 89:9410–9414

    Google Scholar 

  • Yashar BM, Kelley C, Lee K, Errede B, Zon LI (1993) Novel members of the mitogen-activated protein kinase activator family in Xenopus laevis. Mol Cell Biol 13:5738–5748

    Google Scholar 

  • Zhang F, Strand A, Robbins D, Cobb MH, Goldsmith EJ (1994) Atomic structure of the MAP kinase ERK2 at 2.3 Å resolution. Nature 367:704–711

    Google Scholar 

  • Zheng C-F, Guan K-L (1993) Cloning and characterization of two distinct human extracellular signal-regulated kinase activator kinases, MEK1 and MEK2. J Biol Chem 268:11435–11439

    Google Scholar 

  • Zheng C-F, Guan K-L (1994) Activation of MEK family kinases requires phosphorylation of two conserved Ser/Thr residues. EMBO J 13:1123–1131

    Google Scholar 

  • Zhou Z, Gartner A, Cade R, Ammerer G, Errede B (1993) Pheromone-induced signal transduction in Saccharomyces cerevisiae requires the sequential function of three protein kinases. Mol Cell Biol 13:2069–2080

    Google Scholar 

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Authors and Affiliations

  1. Eukaryotic Genetics Group, Biotechnology Research Institute, National Research Council Canada, 6100 Royalmount Ave., H4P 2R2, Montreal, Quebec, Canada

    Karen L. Clark, Daniel Dignard, Robert Larocque, Melanie G. Lee & David Y. Thomas

  2. Dept. of Molecular and Cell Biology, University of Aberdeen, Marischal College, AB9 1AS, Aberdeen, UK

    Pascale J. F. Feldmann & Alistair J. P. Brown

  3. Molecular Sciences, Glaxo Group Research Limited, Greenford Rd., UB6 OHE, Greenford, Middlesex, UK

    Malcolm Whiteway

  4. Dept. of Plant Biology, University of Maryland, 20742, College Park, MD, USA

    Karen L. Clark

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  1. Karen L. Clark
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  2. Pascale J. F. Feldmann
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Communicated by C. P. Hollenberg

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Clark, K.L., Feldmann, P.J.F., Dignard, D. et al. Constitutive activation of the Saccharomyces cerevislae mating response pathway by a MAP kinase kinase from Candida albicans . Molec. Gen. Genet. 249, 609–621 (1995). https://doi.org/10.1007/BF00418030

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