Home LiteratureArticle Details
PMID: 9003780 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Functional analysis of the interaction between the small GTP binding protein Cdc42 and the Ste20 protein kinase in yeast.

The EMBO journal ·Vol. 15 ·No. 24 ·1996-12-16 ·Pages 7046-59

Peter M, Neiman AM, Park HO, van Lohuizen M, Herskowitz I

Abstract

STE20 encodes a protein kinase related to mammalian p65Pak which functions in several signal transduction pathways in yeast, including those involved in pseudohyphal and invasive growth, as well as mating. In addition, Ste20 plays an essential role in cells lacking Cla4, a kinase with significant homology to Ste20. It is not clear how the activity of Ste20 is regulated in response to these different signals in vivo, but it has been demonstrated recently that binding of the small GTP binding protein Cdc42 is able to activate Ste20 in vitro. Here we show that Ste20 functionally interacts with Cdc42 in a GTP-dependent manner in vivo: Ste20 mutants that can no longer bind Cdc42 were unable to restore growth of ste20 cla4 mutant cells. They were also defective for pseudohyphal growth and agar invasion, and displayed reduced mating efficiency when mated with themselves. Surprisingly, however, the kinase activity of such Ste20 mutants was normal when assayed in vitro. Furthermore, these alleles were able to fully activate the MAP kinase pathway triggered by mating pheromones in vivo, suggesting that binding of Cdc42 and Ste20 was not required to activate Ste20. Wild-type Ste20 protein was visualized as a crescent at emerging buds during vegetative growth and at shmoo tips in cells arrested with alpha-factor. In contrast, a Ste20 mutant protein unable to bind Cdc42 was found diffusely throughout the cytoplasm, suggesting that Cdc42 is required to localize Ste20 properly in vivo.

MeSH Terms
Amino Acid Sequence Cell Cycle Proteins/metabolism GTP-Binding Proteins/metabolism Guanosine Triphosphate/metabolism Intracellular Signaling Peptides and Proteins MAP Kinase Kinase Kinases Mating Factor Molecular Sequence Data Peptides/metabolism Pheromones/metabolism Protein Binding Protein Serine-Threonine Kinases/isolation & purification,metabolism Saccharomyces cerevisiae/enzymology,growth & development,metabolism Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid Signal Transduction cdc42 GTP-Binding Protein, Saccharomyces cerevisiae
Chemicals
Cell Cycle Proteins Intracellular Signaling Peptides and Proteins Peptides Pheromones Saccharomyces cerevisiae Proteins Mating Factor Guanosine Triphosphate Protein Serine-Threonine Kinases MAP Kinase Kinase Kinases STE20 protein, S cerevisiae GTP-Binding Proteins cdc42 GTP-Binding Protein, Saccharomyces cerevisiae
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Peter M
Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0448, USA.
Neiman A M
Park H O
van Lohuizen M
Herskowitz I
References (87)
87 references, click to expand
  1. The Rho family GTPases RhoA, Rac1, and CDC42Hs regulate transcriptional activation by SRF.
    Cell. 1995 Jun 30;81(7):1159-70 PMID: 7600583
  2. A mutant of yeast defective in cellular morphogenesis.
    Science. 1978 Jun 9;200(4346):1171-3 PMID: 349694
  3. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  4. Five SWI genes are required for expression of the HO gene in yeast.
    J Mol Biol. 1984 Oct 5;178(4):853-68 PMID: 6436497
  5. Characterization of two members of the rho gene family from the yeast Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1987 Feb;84(3):779-83 PMID: 3543936
  6. Occurrence in Saccharomyces cerevisiae of a gene homologous to the cDNA coding for the alpha subunit of mammalian G proteins.
    Proc Natl Acad Sci U S A. 1987 Apr;84(8):2140-4 PMID: 3031665
  7. The yeast SCG1 gene: a G alpha-like protein implicated in the a- and alpha-factor response pathway.
    Cell. 1987 Sep 25;50(7):1001-10 PMID: 3113738
  8. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  9. The yeast STE12 protein binds to the DNA sequence mediating pheromone induction.
    Proc Natl Acad Sci U S A. 1989 Aug;86(15):5703-7 PMID: 2668945
  10. STE12, a protein involved in cell-type-specific transcription and signal transduction in yeast, is part of protein-DNA complexes.
    Genes Dev. 1989 Sep;3(9):1349-61 PMID: 2558054
  11. Molecular characterization of CDC42, a Saccharomyces cerevisiae gene involved in the development of cell polarity.
    J Cell Biol. 1990 Jul;111(1):143-52 PMID: 2164028
  12. The GTPase superfamily: conserved structure and molecular mechanism.
    Nature. 1991 Jan 10;349(6305):117-27 PMID: 1898771
  13. Mutational analysis of CDC42Sc, a Saccharomyces cerevisiae gene that encodes a putative GTP-binding protein involved in the control of cell polarity.
    Mol Cell Biol. 1991 Jul;11(7):3537-44 PMID: 1904541
  14. A yeast gene (BEM1) necessary for cell polarization whose product contains two SH3 domains.
    Nature. 1992 Mar 5;356(6364):77-9 PMID: 1538785
  15. Unipolar cell divisions in the yeast S. cerevisiae lead to filamentous growth: regulation by starvation and RAS.
    Cell. 1992 Mar 20;68(6):1077-90 PMID: 1547504
  16. Dominant genetics using a yeast genomic library under the control of a strong inducible promoter.
    Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11589-93 PMID: 1454852
  17. The protein kinase homologue Ste20p is required to link the yeast pheromone response G-protein beta gamma subunits to downstream signalling components.
    EMBO J. 1992 Dec;11(13):4815-24 PMID: 1464311
  18. A dominant truncation allele identifies a gene, STE20, that encodes a putative protein kinase necessary for mating in Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):452-6 PMID: 8421676
  19. An osmosensing signal transduction pathway in yeast.
    Science. 1993 Mar 19;259(5102):1760-3 PMID: 7681220
  20. FAR1 links the signal transduction pathway to the cell cycle machinery in yeast.
    Cell. 1993 May 21;73(4):747-60 PMID: 8500168
  21. Signal transduction via the MAP kinases: proceed at your own RSK.
    Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):5889-92 PMID: 8392180
  22. Far1 and Fus3 link the mating pheromone signal transduction pathway to three G1-phase Cdc28 kinase complexes.
    Mol Cell Biol. 1993 Sep;13(9):5659-69 PMID: 8395009
  23. BUD2 encodes a GTPase-activating protein for Bud1/Rsr1 necessary for proper bud-site selection in yeast.
    Nature. 1993 Sep 16;365(6443):269-74 PMID: 8371782
  24. Ste20-like protein kinases are required for normal localization of cell growth and for cytokinesis in budding yeast.
    Genes Dev. 1995 Aug 1;9(15):1817-30 PMID: 7649470
  25. Role for the Rho-family GTPase Cdc42 in yeast mating-pheromone signal pathway.
    Nature. 1995 Aug 24;376(6542):702-5 PMID: 7651520
  26. Association of the yeast pheromone response G protein beta gamma subunits with the MAP kinase scaffold Ste5p.
    Science. 1995 Sep 15;269(5230):1572-5 PMID: 7667635
  27. Pheromone signalling in Saccharomyces cerevisiae requires the small GTP-binding protein Cdc42p and its activator CDC24.
    Mol Cell Biol. 1995 Oct;15(10):5246-57 PMID: 7565673
  28. Activation of the SAPK pathway by the human STE20 homologue germinal centre kinase.
    Nature. 1995 Oct 26;377(6551):750-4 PMID: 7477268
  29. Activation of mitogen-activated protein kinase cascades by p21-activated protein kinases in cell-free extracts of Xenopus oocytes.
    J Biol Chem. 1995 Nov 3;270(44):26067-70 PMID: 7592806
  30. Identification and molecular cloning of a p21cdc42/rac1-activated serine/threonine kinase that is rapidly activated by thrombin in platelets.
    J Biol Chem. 1995 Nov 3;270(44):26690-7 PMID: 7592896
  31. Pheromone response in yeast: association of Bem1p with proteins of the MAP kinase cascade and actin.
    Science. 1995 Nov 17;270(5239):1210-3 PMID: 7502048
  32. Saccharomyces cerevisiae cells execute a default pathway to select a mate in the absence of pheromone gradients.
    J Cell Biol. 1995 Nov;131(4):845-61 PMID: 7490289
  33. FAR1 is required for oriented polarization of yeast cells in response to mating pheromones.
    J Cell Biol. 1995 Nov;131(4):863-73 PMID: 7490290
  34. A conserved binding motif defines numerous candidate target proteins for both Cdc42 and Rac GTPases.
    J Biol Chem. 1995 Dec 8;270(49):29071-4 PMID: 7493928
  35. The Rac and Rho pathways as a source of drug targets for Ras-mediated malignancies.
    Curr Opin Biotechnol. 1995 Dec;6(6):668-74 PMID: 8527838
  36. 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.
    EMBO J. 1995 Dec 1;14(23):5931-8 PMID: 8846785
  37. The SLT2(MPK1) MAP kinase is activated during periods of polarized cell growth in yeast.
    EMBO J. 1996 Jan 2;15(1):83-91 PMID: 8598209
  38. Dynamics and organization of MAP kinase signal pathways.
    Mol Reprod Dev. 1995 Dec;42(4):477-85 PMID: 8607979
  39. Wiskott-Aldrich syndrome protein, a novel effector for the GTPase CDC42Hs, is implicated in actin polymerization.
    Cell. 1996 Mar 8;84(5):723-34 PMID: 8625410
  40. Ste12 and Mcm1 regulate cell cycle-dependent transcription of FAR1.
    Mol Cell Biol. 1996 Jun;16(6):2830-7 PMID: 8649392
  41. Ras2 signals via the Cdc42/Ste20/mitogen-activated protein kinase module to induce filamentous growth in Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1996 May 28;93(11):5352-6 PMID: 8643578
  42. Protein kinase cascades activated by stress and inflammatory cytokines.
    Bioessays. 1996 Jul;18(7):567-77 PMID: 8757935
  43. Genetic relationships between the G protein beta gamma complex, Ste5p, Ste20p and Cdc42p: investigation of effector roles in the yeast pheromone response pathway.
    Genetics. 1996 May;143(1):103-17 PMID: 8722766
  44. Phosphorylation-dependent activation of the Ras-GRF/CDC25Mm exchange factor by muscarinic receptors and G-protein beta gamma subunits.
    Nature. 1996 Jul 18;382(6588):268-72 PMID: 8717044
  45. Human Ste20 homologue hPAK1 links GTPases to the JNK MAP kinase pathway.
    Curr Biol. 1996 May 1;6(5):598-605 PMID: 8805275
  46. Establishment of cell polarity in yeast.
    Cold Spring Harb Symp Quant Biol. 1995;60:729-44 PMID: 8824448
  47. Protein-protein interactions in the yeast pheromone response pathway: Ste5p interacts with all members of the MAP kinase cascade.
    Genetics. 1994 Nov;138(3):609-19 PMID: 7851759
  48. Improved green fluorescence.
    Nature. 1995 Feb 23;373(6516):663-4 PMID: 7854443
  49. Regulation of cortical actin cytoskeleton assembly during polarized cell growth in budding yeast.
    J Cell Biol. 1995 Feb;128(4):599-615 PMID: 7860633
  50. Cell polarization directed by extracellular cues in yeast.
    Mol Biol Cell. 1994 Nov;5(11):1169-75 PMID: 7865882
  51. The Ras-related protein Cdc42Hs and bradykinin promote formation of peripheral actin microspikes and filopodia in Swiss 3T3 fibroblasts.
    Mol Cell Biol. 1995 Apr;15(4):1942-52 PMID: 7891688
  52. Localization of protein kinases by anchoring proteins: a theme in signal transduction.
    Science. 1995 Apr 14;268(5208):247-51 PMID: 7716516
  53. Rho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia.
    Cell. 1995 Apr 7;81(1):53-62 PMID: 7536630
  54. A novel serine kinase activated by rac1/CDC42Hs-dependent autophosphorylation is related to PAK65 and STE20.
    EMBO J. 1995 May 1;14(9):1970-8 PMID: 7744004
  55. Rho-related proteins: actin cytoskeleton and cell cycle.
    Curr Opin Genet Dev. 1995 Feb;5(1):24-30 PMID: 7749321
  56. Regulation of the polarization of T cells toward antigen-presenting cells by Ras-related GTPase CDC42.
    Proc Natl Acad Sci U S A. 1995 May 23;92(11):5027-31 PMID: 7761442
  57. Shk1, a homolog of the Saccharomyces cerevisiae Ste20 and mammalian p65PAK protein kinases, is a component of a Ras/Cdc42 signaling module in the fission yeast Schizosaccharomyces pombe.
    Proc Natl Acad Sci U S A. 1995 Jun 20;92(13):6180-4 PMID: 7597098
  58. The small GTP-binding proteins Rac1 and Cdc42 regulate the activity of the JNK/SAPK signaling pathway.
    Cell. 1995 Jun 30;81(7):1137-46 PMID: 7600581
  59. Polarization of yeast cells in spatial gradients of alpha mating factor.
    Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8332-6 PMID: 8397402
  60. Rab GTPases in vesicular transport.
    Curr Opin Cell Biol. 1993 Aug;5(4):613-20 PMID: 8257602
  61. Elements of the yeast pheromone response pathway required for filamentous growth of diploids.
    Science. 1993 Dec 10;262(5140):1741-4 PMID: 8259520
  62. Control of the yeast bud-site assembly GTPase Cdc42. Catalysis of guanine nucleotide exchange by Cdc24 and stimulation of GTPase activity by Bem3.
    J Biol Chem. 1994 Jan 28;269(4):2369-72 PMID: 8300560
  63. A brain serine/threonine protein kinase activated by Cdc42 and Rac1.
    Nature. 1994 Jan 6;367(6458):40-6 PMID: 8107774
  64. Subcellular localization of Cdc42p, a Saccharomyces cerevisiae GTP-binding protein involved in the control of cell polarity.
    Mol Biol Cell. 1993 Dec;4(12):1307-16 PMID: 8167411
  65. MAP kinase kinase kinase, MAP kinase kinase and MAP kinase.
    Curr Opin Genet Dev. 1994 Feb;4(1):82-9 PMID: 8193545
  66. Sex, stress and integrity: the importance of MAP kinases in yeast.
    Curr Opin Genet Dev. 1994 Feb;4(1):90-5 PMID: 8193546
  67. Requirement for Ras in Raf activation is overcome by targeting Raf to the plasma membrane.
    Nature. 1994 Jun 2;369(6479):411-4 PMID: 8196769
  68. A biochemical function for ras--at last.
    Science. 1994 Jun 3;264(5164):1413-4 PMID: 8197454
  69. Activation of Raf as a result of recruitment to the plasma membrane.
    Science. 1994 Jun 3;264(5164):1463-7 PMID: 7811320
  70. Differential expression of a novel protein kinase in human B lymphocytes. Preferential localization in the germinal center.
    J Biol Chem. 1994 Jun 17;269(24):16802-9 PMID: 7515885
  71. Identification of genes required for normal pheromone-induced cell polarization in Saccharomyces cerevisiae.
    Genetics. 1994 Apr;136(4):1287-96 PMID: 8013906
  72. Complexes between STE5 and components of the pheromone-responsive mitogen-activated protein kinase module.
    Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7762-6 PMID: 8052657
  73. Ste5 tethers multiple protein kinases in the MAP kinase cascade required for mating in S. cerevisiae.
    Cell. 1994 Aug 12;78(3):499-512 PMID: 8062390
  74. Diversity in function and regulation of MAP kinase pathways.
    Trends Biochem Sci. 1994 Jun;19(6):236-40 PMID: 8073500
  75. Cooperative interaction of S. pombe proteins required for mating and morphogenesis.
    Cell. 1994 Oct 7;79(1):131-41 PMID: 7923372
  76. Phosphate-regulated inactivation of the kinase PHO80-PHO85 by the CDK inhibitor PHO81.
    Science. 1994 Oct 7;266(5182):122-6 PMID: 7939631
  77. SMK1, a developmentally regulated MAP kinase, is required for spore wall assembly in Saccharomyces cerevisiae.
    Genes Dev. 1994 Sep 15;8(18):2151-61 PMID: 7958885
  78. Mutation of the SPS1-encoded protein kinase of Saccharomyces cerevisiae leads to defects in transcription and morphology during spore formation.
    Genes Dev. 1994 Sep 15;8(18):2162-75 PMID: 7958886
  79. Interactions between the bud emergence proteins Bem1p and Bem2p and Rho-type GTPases in yeast.
    J Cell Biol. 1994 Dec;127(5):1395-406 PMID: 7962098
  80. Cell polarity in yeast.
    Trends Genet. 1994 Sep;10(9):328-33 PMID: 7974747
  81. Elements of a single MAP kinase cascade in Saccharomyces cerevisiae mediate two developmental programs in the same cell type: mating and invasive growth.
    Genes Dev. 1994 Dec 15;8(24):2974-85 PMID: 8001818
  82. MAP kinase pathways in yeast: for mating and more.
    Cell. 1995 Jan 27;80(2):187-97 PMID: 7834739
  83. Cell division. Septins in common?
    Curr Biol. 1994 Oct 1;4(10):907-10 PMID: 7850425
  84. Molecular characterization of Ste20p, a potential mitogen-activated protein or extracellular signal-regulated kinase kinase (MEK) kinase kinase from Saccharomyces cerevisiae.
    J Biol Chem. 1995 Jul 7;270(27):15984-92 PMID: 7608157
  85. The proliferation of MAP kinase signaling pathways in yeast.
    Curr Opin Cell Biol. 1995 Apr;7(2):197-202 PMID: 7612271
  86. Regulation of human leukocyte p21-activated kinases through G protein--coupled receptors.
    Science. 1995 Jul 14;269(5221):221-3 PMID: 7618083
  87. Selective activation of the JNK signaling cascade and c-Jun transcriptional activity by the small GTPases Rac and Cdc42Hs.
    Cell. 1995 Jun 30;81(7):1147-57 PMID: 7600582
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1996-12-16
Pages
7046-59
Language
English
Region
England
NLM ID
8208664
PMCID
PMC452530
Subset
IM
Grants
NIGMS NIH HHS · GM48052 · United States
External Links
PubMed source PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2