Differential Stabilities of Phosphorylated Response Regulator Domains Reflect Functional Roles of the Yeast Osmoregulatory SLN1 and SSK1 Proteins

ABSTRACT Osmoregulation in Saccharomyces cerevisiae involves a multistep phosphorelay system requiring three proteins, SLN1, YPD1, and SSK1, that are related to bacterial two-component signaling proteins, in particular, those involved in regulating sporulation inBacillus subtilis and anaerobic respiration inEscherichia coli. The SLN1-YPD1-SSK1 phosphorelay regulates a downstream mitogen-activated protein kinase cascade which ultimately controls the concentration of glycerol within the cell under hyperosmotic stress conditions. The C-terminal response regulator domains of SLN1 and SSK1 and full-length YPD1 have been overexpressed and purified from E. coli. A heterologous system consisting of acetyl phosphate, the bacterial chemotaxis response regulator CheY, and YPD1 has been developed as an efficient means of phosphorylating SLN1 and SSK1 in vitro. The homologous regulatory domains of SLN1 and SSK1 exhibit remarkably different phosphorylated half-lives, a finding that provides insight into the distinct roles that these phosphorylation-dependent regulatory domains play in the yeast osmosensory signal transduction pathway.

[1]  T. Mizuno,et al.  Evidence for the physiological importance of the phosphotransfer between the two regulatory components, EnvZ and OmpR, in osmoregulation in Escherichia coli. , 1989, The Journal of biological chemistry.

[2]  B. Morgan,et al.  The Mcs4 response regulator coordinately controls the stress-activated Wak1-Wis1-Sty1 MAP kinase pathway and fission yeast cell cycle. , 1997, Genes & development.

[3]  E. Meyerowitz,et al.  ETR2 is an ETR1-like gene involved in ethylene signaling in Arabidopsis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[4]  D. Koshland.  Effect of Catalysts on the Hydrolysis of Acetyl Phosphate. Nucleophilic Displacement Mechanisms in Enzymatic Reactions1 , 1952 .

[5]  J. Hoch,et al.  A phosphotransferase activity of the Bacillus subtilis sporulation protein Spo0F that employs phosphoramidate substrates. , 1996, Biochemistry.

[6]  T. Mizuno,et al.  Response regulators implicated in His-to-Asp phosphotransfer signaling in Arabidopsis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[7]  J. Stock,et al.  Acetyl phosphate and the activation of two-component response regulators. , 1994, The Journal of biological chemistry.

[8]  H. Ruis,et al.  The HOG pathway controls osmotic regulation of transcription via the stress response element (STRE) of the Saccharomyces cerevisiae CTT1 gene. , 1994, The EMBO journal.

[9]  T. Mizuno,et al.  A novel device of bacterial signal transducers. , 1994, The EMBO journal.

[10]  T. Maeda,et al.  Activation of yeast PBS2 MAPKK by MAPKKKs or by binding of an SH3-containing osmosensor. , 1995, Science.

[11]  S. Iuchi,et al.  Phosphorylation/dephosphorylation of the receiver module at the conserved aspartate residue controls transphosphorylation activity of histidine kinase in sensor protein ArcB of Escherichia coli. , 1993, The Journal of biological chemistry.

[12]  R C Stewart,et al.  Yeast Skn7p functions in a eukaryotic two‐component regulatory pathway. , 1994, The EMBO journal.

[13]  M. Zinda,et al.  The hybrid histidine kinase dhkB regulates spore germination in Dictyostelium discoideum. , 1998, Developmental biology.

[14]  A. Clarke,et al.  A method for the separation of GST fusion proteins from co-purifying GroEL. , 1996, Trends in genetics : TIG.

[15]  E. Stadtman.  [39] Preparation and assay of acetyl phosphate , 1957 .

[16]  Tatsuya Maeda,et al.  A two-component system that regulates an osmosensing MAP kinase cascade in yeast , 1994, Nature.

[17]  I. Ota,et al.  A yeast protein similar to bacterial two-component regulators. , 1993, Science.

[18]  T. Kawamoto,et al.  Signal transduction in the phosphate regulon of Escherichia coli involves phosphotransfer between PhoR and PhoB proteins. , 1989, Journal of molecular biology.

[19]  L. Johnston,et al.  Two-component signal-transduction systems in budding yeast MAP a different pathway? , 1995, Trends in cell biology.

[20]  E. Meyerowitz,et al.  Ethylene insensitivity conferred by Arabidopsis ERS gene. , 1995, Science.

[21]  J. Stock,et al.  Phosphorylation of bacterial response regulator proteins by low molecular weight phospho-donors. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[22]  P. Newell,et al.  Evidence that the RdeA protein is a component of a multistep phosphorelay modulating rate of development in Dictyostelium , 1998, The EMBO journal.

[23]  Kenji Oosawa,et al.  Phosphorylation of three proteins in the signaling pathway of bacterial chemotaxis , 1988, Cell.

[24]  Francesc Posas,et al.  Yeast HOG1 MAP Kinase Cascade Is Regulated by a Multistep Phosphorelay Mechanism in the SLN1–YPD1–SSK1 “Two-Component” Osmosensor , 1996, Cell.

[25]  A. Ninfa,et al.  Role of phosphorylated metabolic intermediates in the regulation of glutamine synthetase synthesis in Escherichia coli , 1992, Journal of bacteriology.

[26]  Barry L. Wanner,et al.  Kinetic Comparison of the Specificity of the Vancomycin Resistance Kinase VanS for Two Response Regulators, VanR and PhoB† , 1996 .

[27]  Hsiao-Ching Yen,et al.  An Ethylene-Inducible Component of Signal Transduction Encoded by Never-ripe , 1995, Science.

[28]  J. S. Parkinson,et al.  Signal Transduction via the Multi-Step Phosphorelay: Not Necessarily a Road Less Traveled , 1996, Cell.

[29]  A. Ninfa,et al.  Phosphorylation and dephosphorylation of a bacterial transcriptional activator by a transmembrane receptor. , 1989, Genes & development.

[30]  K. Shinozaki,et al.  Stress‐responsive expression of genes for two‐component response regulator‐like proteins in Arabidopsis thaliana , 1998, FEBS letters.

[31]  W. H. Mager,et al.  Osmostress response of the yeast Saccharomyces. , 1993, Molecular microbiology.

[32]  H. Saito,et al.  Two-component signal transducers and MAPK cascades. , 1997, Trends in biochemical sciences.

[33]  A. Blomberg,et al.  Roles of glycerol and glycerol-3-phosphate dehydrogenase (NAD+) in acquired osmotolerance of Saccharomyces cerevisiae , 1989, Journal of bacteriology.

[34]  C. Schutt,et al.  Three-dimensional structure of CheY, the response regulator of bacterial chemotaxis , 1989, Nature.

[35]  J. Stock,et al.  Sensory transduction in bacterial chemotaxis involves phosphotransfer between Che proteins. , 1988, Biochemical and biophysical research communications.

[36]  C. Malone,et al.  Activated Alleles of Yeast SLN1 Increase Mcm1-dependent Reporter Gene Expression and Diminish Signaling through the Hog1 Osmosensing Pathway* , 1997, The Journal of Biological Chemistry.

[37]  G. Shaulsky,et al.  A two‐component histidine kinase gene that functions in Dictyostelium development. , 1996, The EMBO journal.

[38]  Takeshi Mizuno,et al.  Insights into Multistep Phosphorelay from the Crystal Structure of the C-Terminal HPt Domain of ArcB , 1997, Cell.

[39]  M. Simon,et al.  Hyphal development in Neurospora crassa: involvement of a two-component histidine kinase. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[40]  Francesc Posas,et al.  Activation of the yeast SSK2 MAP kinase kinase kinase by the SSK1 two‐component response regulator , 1998, The EMBO journal.

[41]  Stefan Dipl.-Ing. Schuster,et al.  The hybrid histidine kinase DokA is part of the osmotic response system of Dictyostelium. , 1996, The EMBO journal.

[42]  J. Hoch,et al.  Initiation of sporulation in B. subtilis is controlled by a multicomponent phosphorelay , 1991, Cell.

[43]  J M Thevelein,et al.  GPD1, which encodes glycerol-3-phosphate dehydrogenase, is essential for growth under osmotic stress in Saccharomyces cerevisiae, and its expression is regulated by the high-osmolarity glycerol response pathway , 1994, Molecular and cellular biology.

[44]  T. Mizuno,et al.  Phosphotransfer circuitry of the putative multi‐signal transducer, ArcB, of Escherichia coli: in vitro studies with mutants , 1995, Molecular microbiology.

[45]  Stefan Dipl.-Ing. Schuster,et al.  Phosphorylation in halobacterial signal transduction. , 1995, The EMBO journal.

[46]  A. West,et al.  Purification, crystallization and preliminary X-ray diffraction analysis of the yeast phosphorelay protein YPD1. , 1999, Acta crystallographica. Section D, Biological crystallography.

[47]  R. Gunsalus,et al.  Phosphorylation and dephosphorylation of the NarQ, NarX, and NarL proteins of the nitrate-dependent two-component regulatory system of Escherichia coli , 1994, Journal of bacteriology.

[48]  T. Kakimoto.  CKI1, a Histidine Kinase Homolog Implicated in Cytokinin Signal Transduction , 1996, Science.

[49]  J. L. Brown,et al.  Yeast Skn7p activity is modulated by the Sln1p-Ypd1p osmosensor and contributes to regulation of the HOG pathway , 1998, Molecular and General Genetics MGG.

[50]  Jeff F. Miller,et al.  Integration of multiple domains in a two‐component sensor protein: the Bordetella pertussis BvgAS phosphorelay. , 1996, The EMBO journal.

[51]  R. Smith,et al.  Techniques in the detection and characterization of phosphoramidate-containing proteins. , 1984, Methods in enzymology.

[52]  C. Walsh,et al.  Purification and characterization of VanR and the cytosolic domain of VanS: a two-component regulatory system required for vancomycin resistance in Enterococcus faecium BM4147. , 1993, Biochemistry.

[53]  R. Fleischmann,et al.  The complete genome sequence of the hyperthermophilic, sulphate-reducing archaeon Archaeoglobus fulgidus , 1997, Nature.

[54]  J. Hoch,et al.  A source of response regulator autophosphatase activity: the critical role of a residue adjacent to the Spo0F autophosphorylation active site. , 1998, Biochemistry.

[55]  G. Church,et al.  Complete genome sequence of Methanobacterium thermoautotrophicum deltaH: functional analysis and comparative genomics , 1997, Journal of bacteriology.

[56]  E. Meyerowitz,et al.  Arabidopsis ethylene-response gene ETR1: similarity of product to two-component regulators. , 1993, Science.