PDK1 Homologs Activate the Pkc1–Mitogen-Activated Protein Kinase Pathway in Yeast

ABSTRACT PDK1 (phosphoinositide-dependent kinase 1) is a mammalian growth factor-regulated serine/threonine kinase. Using a genetic selection based on a mutant form of the yeast MAP kinase kinase Ste7, we isolated a gene, PKH2, encoding a structurally and functionally conserved yeast homolog of PDK1. Yeast cells lacking bothPKH2 and PKH1, encoding another PDK1 homolog, were nonviable, indicating that Pkh1 and Pkh2 share an essential function. A temperature-sensitive mutant, pkh1D398Gpkh2, was phenotypically similar to mutants defective in the Pkc1–mitogen-activated protein kinase (MAPK) pathway. Genetic epistasis analyses, the phosphorylation of Pkc1 by Pkh2 in vitro, and reduced Pkc1 activity in the pkh1D398G pkh2mutant indicate that Pkh functions upstream of Pkc1. The Pkh2 phosphorylation site in Pkc1 (Thr-983) is part of a conserved PDK1 target motif and essential for Pkc1 function. Thus, the yeast PDK1 homologs activate Pkc1 and the Pkc1-effector MAPK pathway.

[1]  J. Thorner,et al.  Functional counterparts of mammalian protein kinases PDK1 and SGK in budding yeast , 1999, Current Biology.

[2]  M. Gustin,et al.  MAP Kinase Pathways in the YeastSaccharomyces cerevisiae , 1998, Microbiology and Molecular Biology Reviews.

[3]  Y. Ohya,et al.  The Rho1 effector Pkc1, but not Bni1, mediates signalling from Tor2 to the actin cytoskeleton , 1998, Current Biology.

[4]  Philip R. Cohen,et al.  Protein kinase C isotypes controlled by phosphoinositide 3-kinase through the protein kinase PDK1. , 1998, Science.

[5]  F. Banuett.  Signalling in the Yeasts: An Informational Cascade with Links to the Filamentous Fungi , 1998, Microbiology and Molecular Biology Reviews.

[6]  Y. Ip,et al.  Signal transduction by the c-Jun N-terminal kinase (JNK)--from inflammation to development. , 1998, Current opinion in cell biology.

[7]  F. Posas,et al.  Signal transduction by MAP kinase cascades in budding yeast. , 1998, Current opinion in microbiology.

[8]  F. McCormick,et al.  Protein kinase B kinases that mediate phosphatidylinositol 3,4,5-trisphosphate-dependent activation of protein kinase B. , 1998, Science.

[9]  M. Andjelkovic,et al.  Phosphorylation and activation of p70s6k by PDK1. , 1998, Science.

[10]  Dario R. Alessi,et al.  3-Phosphoinositide-dependent protein kinase 1 (PDK1) phosphorylates and activates the p70 S6 kinase in vivo and in vitro , 1998, Current Biology.

[11]  Y. Watanabe,et al.  Characterization of a serum response factor-like protein in Saccharomyces cerevisiae, Rlm1, which has transcriptional activity regulated by the Mpk1 (Slt2) mitogen-activated protein kinase pathway , 1997, Molecular and cellular biology.

[12]  P. Cohen,et al.  Characterization of a 3-phosphoinositide-dependent protein kinase which phosphorylates and activates protein kinase Bα , 1997, Current Biology.

[13]  M. Cobb,et al.  Mitogen-activated protein kinase pathways. , 1997, Current opinion in cell biology.

[14]  N. Hay,et al.  The PI 3-kinase/Akt signaling pathway delivers an anti-apoptotic signal. , 1997, Genes & development.

[15]  G. Evan,et al.  Suppression of c-Myc-induced apoptosis by Ras signalling through PI(3)K and PKB , 1997, Nature.

[16]  M. Whiteway,et al.  Pheromone signalling and polarized morphogenesis in yeast. , 1997, Current opinion in genetics & development.

[17]  David R. Kaplan,et al.  Regulation of Neuronal Survival by the Serine-Threonine Protein Kinase Akt , 1997, Science.

[18]  P. Cohen,et al.  Mechanism of activation of protein kinase B by insulin and IGF‐1. , 1996, The EMBO journal.

[19]  M. Hall,et al.  TOR2 is required for organization of the actin cytoskeleton in yeast. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[20]  J. Avruch,et al.  Protein kinase cascades activated by stress and inflammatory cytokines , 1996, BioEssays : news and reviews in molecular, cellular and developmental biology.

[21]  Y. Anraku,et al.  Activation of Yeast Protein Kinase C by Rho1 GTPase (*) , 1996, The Journal of Biological Chemistry.

[22]  K. Irie,et al.  Identification of a Member of the MAPKKK Family as a Potential Mediator of TGF-β Signal Transduction , 1995, Science.

[23]  K. Tanaka,et al.  A downstream target of RHO1 small GTP‐binding protein is PKC1, a homolog of protein kinase C, which leads to activation of the MAP kinase cascade in Saccharomyces cerevisiae. , 1995, The EMBO journal.

[24]  Brian J. Stevenson,et al.  Yeast MEK-dependent signal transduction: response thresholds and parameters affecting fidelity , 1995, Molecular and cellular biology.

[25]  Y. Watanabe,et al.  Yeast RLM1 encodes a serum response factor-like protein that may function downstream of the Mpk1 (Slt2) mitogen-activated protein kinase pathway , 1995, Molecular and cellular biology.

[26]  I. Herskowitz.  MAP kinase pathways in yeast: For mating and more , 1995, Cell.

[27]  B. Errede,et al.  The proliferation of MAP kinase signaling pathways in yeast. , 1995, Current opinion in cell biology.

[28]  A. Newton,et al.  Requirement for negative charge on "activation loop" of protein kinase C. , 1994, The Journal of biological chemistry.

[29]  K. Irie,et al.  Stimulatory effects of yeast and mammalian 14-3-3 proteins on the Raf protein kinase. , 1994, Science.

[30]  H. Riezman,et al.  The END3 gene encodes a protein that is required for the internalization step of endocytosis and for actin cytoskeleton organization in yeast , 1994, Molecular biology of the cell.

[31]  P. Parker,et al.  Threonine-497 is a critical site for permissive activation of protein kinase C alpha. , 1994, The Biochemical journal.

[32]  D. E. Levin,et al.  Saccharomyces cerevisiae PKC1 encodes a protein kinase C (PKC) homolog with a substrate specificity similar to that of mammalian PKC. , 1994, The Journal of biological chemistry.

[33]  C. Mazzoni,et al.  The SLT2 (MPK1) MAP kinase homolog is involved in polarized cell growth in Saccharomyces cerevisiae , 1993, The Journal of cell biology.

[34]  E. Elion,et al.  FUS3 phosphorylates multiple components of the mating signal transduction cascade: evidence for STE12 and FAR1. , 1993, Molecular biology of the cell.

[35]  B. Errede,et al.  Constitutive mutants of the protein kinase STE11 activate the yeast pheromone response pathway in the absence of the G protein. , 1992, Genes & development.

[36]  M. Snyder,et al.  A synthetic lethal screen identifies SLK1, a novel protein kinase homolog implicated in yeast cell morphogenesis and cell growth , 1992, Molecular and cellular biology.

[37]  D. E. Levin,et al.  Mutants in the S. cerevisiae PKC1 gene display a cell cycle-specific osmotic stability defect , 1992, The Journal of cell biology.

[38]  R. Parker,et al.  A rapid method for localized mutagenesis of yeast genes , 1992, Yeast.

[39]  D. E. Levin,et al.  Dominant mutations in a gene encoding a putative protein kinase (BCK1) bypass the requirement for a Saccharomyces cerevisiae protein kinase C homolog , 1992, Molecular and cellular biology.

[40]  J. Small.  The actin cytoskeleton. , 1988, Electron Microscopy Reviews.

[41]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[42]  R. Rothstein.  One-step gene disruption in yeast. , 1983, Methods in enzymology.

[43]  E. Cabib,et al.  Simple and sensitive procedure for screening yeast mutants that lyse at nonpermissive temperatures , 1975, Journal of bacteriology.