Home LiteratureArticle Details
PMID: 10562285 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

PAK-family kinases regulate cell and actin polarization throughout the cell cycle of Saccharomyces cerevisiae.

The Journal of cell biology ·Vol. 147 ·No. 4 ·1999-11-15 ·Pages 845-56

Holly SP, Blumer KJ

Abstract

During the cell cycle of the yeast Saccharomyces cerevisiae, the actin cytoskeleton and cell surface growth are polarized, mediating bud emergence, bud growth, and cytokinesis. We have determined whether p21-activated kinase (PAK)-family kinases regulate cell and actin polarization at one or several points during the yeast cell cycle. Inactivation of the PAK homologues Ste20 and Cla4 at various points in the cell cycle resulted in loss of cell and actin cytoskeletal polarity, but not in depolymerization of F-actin. Loss of PAK function in G1 depolarized the cortical actin cytoskeleton and blocked bud emergence, but allowed isotropic growth and led to defects in septin assembly, indicating that PAKs are effectors of the Rho-guanosine triphosphatase Cdc42. PAK inactivation in S/G2 resulted in depolarized growth of the mother and bud and a loss of actin polarity. Loss of PAK function in mitosis caused a defect in cytokinesis and a failure to polarize the cortical actin cytoskeleton to the mother-bud neck. Cla4-green fluorescent protein localized to sites where the cortical actin cytoskeleton and cell surface growth are polarized, independently of an intact actin cytoskeleton. Thus, PAK family kinases are primary regulators of cell and actin cytoskeletal polarity throughout most or all of the yeast cell cycle. PAK-family kinases in higher organisms may have similar functions.

MeSH Terms
Actins/metabolism Cell Cycle/physiology Cell Polarity/physiology Fungal Proteins/metabolism Genotype Intracellular Signaling Peptides and Proteins Kinetics MAP Kinase Kinase Kinases Microscopy, Video Mitosis Protein Serine-Threonine Kinases/genetics,metabolism Recombinant Fusion Proteins/metabolism Saccharomyces cerevisiae/cytology,genetics,physiology Saccharomyces cerevisiae Proteins cdc42 GTP-Binding Protein/metabolism
Chemicals
Actins Fungal Proteins Intracellular Signaling Peptides and Proteins Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins CLA4 protein, S cerevisiae Protein Serine-Threonine Kinases MAP Kinase Kinase Kinases STE20 protein, S cerevisiae cdc42 GTP-Binding Protein
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Holly S P
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Blumer K J
References (61)
61 references, click to expand
  1. 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
  2. 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
  3. Movement of cortical actin patches in yeast.
    J Cell Biol. 1996 Mar;132(5):861-70 PMID: 8603918
  4. Origins of cell polarity.
    Cell. 1996 Feb 9;84(3):335-44 PMID: 8608587
  5. Movement of yeast cortical actin cytoskeleton visualized in vivo.
    Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):3886-91 PMID: 8632984
  6. Synthetic lethality screen identifies a novel yeast myosin I gene (MYO5): myosin I proteins are required for polarization of the actin cytoskeleton.
    J Cell Biol. 1996 Jun;133(6):1277-91 PMID: 8682864
  7. TOR2 is required for organization of the actin cytoskeleton in yeast.
    Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13780-5 PMID: 8943012
  8. Functional analysis of the interaction between the small GTP binding protein Cdc42 and the Ste20 protein kinase in yeast.
    EMBO J. 1996 Dec 16;15(24):7046-59 PMID: 9003780
  9. Functional characterization of the Cdc42p binding domain of yeast Ste20p protein kinase.
    EMBO J. 1997 Jan 2;16(1):83-97 PMID: 9009270
  10. Bee1, a yeast protein with homology to Wiscott-Aldrich syndrome protein, is critical for the assembly of cortical actin cytoskeleton.
    J Cell Biol. 1997 Feb 10;136(3):649-58 PMID: 9024694
  11. Human p21-activated kinase (Pak1) regulates actin organization in mammalian cells.
    Curr Biol. 1997 Mar 1;7(3):202-10 PMID: 9395435
  12. High rates of actin filament turnover in budding yeast and roles for actin in establishment and maintenance of cell polarity revealed using the actin inhibitor latrunculin-A.
    J Cell Biol. 1997 Apr 21;137(2):399-416 PMID: 9128251
  13. The complex containing actin-related proteins Arp2 and Arp3 is required for the motility and integrity of yeast actin patches.
    Curr Biol. 1997 Jul 1;7(7):519-29 PMID: 9210376
  14. Mitosis in living budding yeast: anaphase A but no metaphase plate.
    Science. 1997 Jul 25;277(5325):574-8 PMID: 9228009
  15. Cla4p, a Saccharomyces cerevisiae Cdc42p-activated kinase involved in cytokinesis, is activated at mitosis.
    Mol Cell Biol. 1997 Sep;17(9):5067-76 PMID: 9271384
  16. Post-Golgi biosynthetic trafficking.
    J Cell Sci. 1997 Dec;110 ( Pt 24):3001-9 PMID: 9365270
  17. The Cdc42 GTPase-associated proteins Gic1 and Gic2 are required for polarized cell growth in Saccharomyces cerevisiae.
    Genes Dev. 1997 Nov 15;11(22):2958-71 PMID: 9367979
  18. Novel Cdc42-binding proteins Gic1 and Gic2 control cell polarity in yeast.
    Genes Dev. 1997 Nov 15;11(22):2972-82 PMID: 9367980
  19. An IQGAP-related protein controls actin-ring formation and cytokinesis in yeast.
    Curr Biol. 1997 Dec 1;7(12):921-9 PMID: 9382845
  20. The phosphorylation site for Ste20p-like protein kinases is essential for the function of myosin-I in yeast.
    J Biol Chem. 1997 Dec 5;272(49):30623-6 PMID: 9388196
  21. Role of the yeast Gin4p protein kinase in septin assembly and the relationship between septin assembly and septin function.
    J Cell Biol. 1998 Nov 2;143(3):719-36 PMID: 9813093
  22. PAK4, a novel effector for Cdc42Hs, is implicated in the reorganization of the actin cytoskeleton and in the formation of filopodia.
    EMBO J. 1998 Nov 16;17(22):6527-40 PMID: 9822598
  23. The Arp2/3 complex mediates actin polymerization induced by the small GTP-binding protein Cdc42.
    Proc Natl Acad Sci U S A. 1998 Dec 22;95(26):15362-7 PMID: 9860974
  24. Tropomyosin-containing actin cables direct the Myo2p-dependent polarized delivery of secretory vesicles in budding yeast.
    J Cell Biol. 1998 Dec 28;143(7):1931-45 PMID: 9864365
  25. Nim1-related kinases coordinate cell cycle progression with the organization of the peripheral cytoskeleton in yeast.
    Genes Dev. 1999 Jan 15;13(2):176-87 PMID: 9925642
  26. The multiple roles of Cyk1p in the assembly and function of the actomyosin ring in budding yeast.
    Mol Biol Cell. 1999 Feb;10(2):283-96 PMID: 9950677
  27. Cdc42: An essential Rho-type GTPase controlling eukaryotic cell polarity.
    Microbiol Mol Biol Rev. 1999 Mar;63(1):54-105 PMID: 10066831
  28. The interaction between N-WASP and the Arp2/3 complex links Cdc42-dependent signals to actin assembly.
    Cell. 1999 Apr 16;97(2):221-31 PMID: 10219243
  29. Membrane targeting of p21-activated kinase 1 (PAK1) induces neurite outgrowth from PC12 cells.
    EMBO J. 1998 Feb 2;17(3):754-64 PMID: 9451000
  30. A conserved negative regulatory region in alphaPAK: inhibition of PAK kinases reveals their morphological roles downstream of Cdc42 and Rac1.
    Mol Cell Biol. 1998 Apr;18(4):2153-63 PMID: 9528787
  31. Plasma membrane localization is required for RGS4 function in Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1998 May 12;95(10):5584-9 PMID: 9576926
  32. Spatial regulation of exocytosis: lessons from yeast.
    J Cell Biol. 1998 Aug 10;142(3):609-12 PMID: 9700152
  33. Involvement of an actomyosin contractile ring in Saccharomyces cerevisiae cytokinesis.
    J Cell Biol. 1998 Sep 7;142(5):1301-12 PMID: 9732290
  34. Actin cytoskeleton organization regulated by the PAK family of protein kinases.
    Curr Biol. 1998 Aug 27;8(17):967-70 PMID: 9742399
  35. A morphogenesis checkpoint monitors the actin cytoskeleton in yeast.
    J Cell Biol. 1998 Sep 21;142(6):1487-99 PMID: 9744879
  36. Assembly and function of the actin cytoskeleton of yeast: relationships between cables and patches.
    J Cell Biol. 1998 Sep 21;142(6):1501-17 PMID: 9744880
  37. Control of mitotic events by the Cdc42 GTPase, the Clb2 cyclin and a member of the PAK kinase family.
    Curr Biol. 1998 Sep 10;8(18):991-1000 PMID: 9740799
  38. The septins are required for the mitosis-specific activation of the Gin4 kinase.
    J Cell Biol. 1998 Nov 2;143(3):709-17 PMID: 9813092
  39. Secretion and cell-surface growth are blocked in a temperature-sensitive mutant of Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1979 Apr;76(4):1858-62 PMID: 377286
  40. Localized secretion of acid phosphatase reflects the pattern of cell surface growth in Saccharomyces cerevisiae.
    J Cell Biol. 1980 Jul;86(1):123-8 PMID: 6998984
  41. Two nuclear mutations that block mitochondrial protein import in yeast.
    Proc Natl Acad Sci U S A. 1984 Aug;81(15):4819-23 PMID: 6235522
  42. Phenotypic analysis of temperature-sensitive yeast actin mutants.
    Cell. 1985 Feb;40(2):405-16 PMID: 3967297
  43. Immunofluorescence localization of the Saccharomyces cerevisiae CDC12 gene product to the vicinity of the 10-nm filaments in the mother-bud neck.
    Mol Cell Biol. 1987 Oct;7(10):3678-87 PMID: 3316985
  44. The RAD9 gene controls the cell cycle response to DNA damage in Saccharomyces cerevisiae.
    Science. 1988 Jul 15;241(4863):317-22 PMID: 3291120
  45. CDC42 and CDC43, two additional genes involved in budding and the establishment of cell polarity in the yeast Saccharomyces cerevisiae.
    J Cell Biol. 1990 Jul;111(1):131-42 PMID: 2195038
  46. 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
  47. Cellular morphogenesis in the Saccharomyces cerevisiae cell cycle: localization of the CDC3 gene product and the timing of events at the budding site.
    J Cell Biol. 1991 Feb;112(4):535-44 PMID: 1993729
  48. Getting started with yeast.
    Methods Enzymol. 1991;194:3-21 PMID: 2005794
  49. Immunofluorescence methods for yeast.
    Methods Enzymol. 1991;194:565-602 PMID: 2005809
  50. A yeast gene (BEM1) necessary for cell polarization whose product contains two SH3 domains.
    Nature. 1992 Mar 5;356(6364):77-9 PMID: 1538785
  51. Effects of null mutations and overexpression of capping protein on morphogenesis, actin distribution and polarized secretion in yeast.
    J Cell Biol. 1992 Dec;119(5):1151-62 PMID: 1447293
  52. Morphogenesis in the yeast cell cycle: regulation by Cdc28 and cyclins.
    J Cell Biol. 1993 Mar;120(6):1305-20 PMID: 8449978
  53. Cell cycle arrest of cdc mutants and specificity of the RAD9 checkpoint.
    Genetics. 1993 May;134(1):63-80 PMID: 8514150
  54. Heat-inducible degron: a method for constructing temperature-sensitive mutants.
    Science. 1994 Mar 4;263(5151):1273-6 PMID: 8122109
  55. What are the basic functions of microfilaments? Insights from studies in budding yeast.
    J Cell Biol. 1994 Aug;126(4):821-5 PMID: 8051208
  56. Mitotic checkpoint genes in budding yeast and the dependence of mitosis on DNA replication and repair.
    Genes Dev. 1994 Mar 15;8(6):652-65 PMID: 7926756
  57. Regulation of cortical actin cytoskeleton assembly during polarized cell growth in budding yeast.
    J Cell Biol. 1995 Feb;128(4):599-615 PMID: 7860633
  58. Identification and molecular characterization of a yeast myosin I.
    Cell Motil Cytoskeleton. 1995;30(1):73-84 PMID: 7728870
  59. A cell cycle checkpoint monitors cell morphogenesis in budding yeast.
    J Cell Biol. 1995 May;129(3):739-49 PMID: 7730408
  60. Cell cycle control of morphogenesis in budding yeast.
    Curr Opin Genet Dev. 1995 Feb;5(1):17-23 PMID: 7749320
  61. Actin filaments in yeast are unstable in the absence of capping protein or fimbrin.
    J Cell Biol. 1995 Dec;131(6 Pt 1):1483-93 PMID: 8522605
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.147.4.845
Published
1999-11-15
Pages
845-56
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2156167
Subset
IM
Grants
NIGMS NIH HHS · GM44592 · United States
External Links
PubMed source DOI: 10.1083/jcb.147.4.845 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: [email protected]

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