Home LiteratureArticle Details
PMID: 7744004 Published · ppublish English Comparative Study Journal Article

A novel serine kinase activated by rac1/CDC42Hs-dependent autophosphorylation is related to PAK65 and STE20.

The EMBO journal ·Vol. 14 ·No. 9 ·1995-05-01 ·Pages 1970-8

Martin GA, Bollag G, McCormick F, Abo A

Abstract

We identified three proteins in neutrophil cytosol of molecular size 65, 62 and 68 kDa which interact in a GTP-dependent manner with rac1 and CDC42Hs, but not with rho. Purification of p65 and subsequent peptide sequencing revealed identity to rat brain PAK65 and to yeast STE20 kinase domains. Based on these sequences we screened a human placenta library and cloned the full-length cDNA. The complete amino acid sequence of the human cDNA shares approximately identity with rat brain PAK65; within the kinase domain the human protein shares > 95% and approximately 63% identity with rat PAK65 and yeast STE20 respectively. The new human (h)PAK65 mRNA is ubiquitously expressed and hPAK65 protein is distinct from either human or rat brain PAK65. Recombinant hPAK65 exhibits identical specificity to the endogenous p65; both can bind rac1 and CDC42Hs in a GTP-dependent manner. The GTP-bound forms of rac1 and CDC42Hs induce autophosphorylation of hPAK65 on serine residues only. hPAK65 activated by either rac1 or CDC42Hs is phosphorylated on the same sites. Induction of hPAK65 autophosphorylation by rac1 or CDC42Hs stimulates hPAK65 kinase activity towards myelin basic protein and once hPAK65 is activated, rac1 or CDC42Hs are no longer required to keep it active. The affinities of rac/CDC42Hs for the non-phosphorylated and phosphorylated hPAK65 were similar. hPAK65 had only a marginal effect on the intrinsic GTPase activity of CDC42Hs, but significantly affected the binding and GAP activity of p190. These data are consistent with a model in which hPAK65 functions as an effector molecule for rac1 and CDC42Hs.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cell Cycle Proteins/metabolism Cell Line Cloning, Molecular DNA, Complementary/genetics Enzyme Activation GTP-Binding Proteins/metabolism Humans Intracellular Signaling Peptides and Proteins MAP Kinase Kinase Kinases Molecular Sequence Data Neutrophils/metabolism Phosphorylation Protein Serine-Threonine Kinases/genetics,metabolism Rats Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid Tissue Distribution cdc42 GTP-Binding Protein rac GTP-Binding Proteins
Chemicals
Cell Cycle Proteins DNA, Complementary Intracellular Signaling Peptides and Proteins Saccharomyces cerevisiae Proteins Protein Serine-Threonine Kinases MAP Kinase Kinase Kinases STE20 protein, S cerevisiae GTP-Binding Proteins cdc42 GTP-Binding Protein rac GTP-Binding Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Martin G A
Onyx Pharmaceuticals, Richmond, CA 94806, USA.
Bollag G
McCormick F
Abo A
References (33)
33 references, click to expand
  1. MAP kinase-related FUS3 from S. cerevisiae is activated by STE7 in vitro.
    Nature. 1993 Mar 18;362(6417):261-4 PMID: 8384702
  2. Inhibition of PMA-induced, LFA-1-dependent lymphocyte aggregation by ADP ribosylation of the small molecular weight GTP binding protein, rho.
    J Cell Biol. 1993 Mar;120(6):1529-37 PMID: 7680658
  3. Tyrosine kinase-stimulated guanine nucleotide exchange activity of Vav in T cell activation.
    Science. 1993 May 7;260(5109):822-5 PMID: 8484124
  4. The biochemical basis of the NADPH oxidase of phagocytes.
    Trends Biochem Sci. 1993 Feb;18(2):43-7 PMID: 8488557
  5. Accelerated high-sensitivity microsequencing of proteins and peptides using a miniature reaction cartridge.
    Protein Sci. 1992 Sep;1(9):1215-24 PMID: 1304398
  6. A conserved kinase cascade for MAP kinase activation in yeast.
    Curr Opin Cell Biol. 1993 Apr;5(2):254-60 PMID: 8389568
  7. Activation of platelet phosphatidylinositide 3-kinase requires the small GTP-binding protein Rho.
    J Biol Chem. 1993 Oct 25;268(30):22251-4 PMID: 8226730
  8. Cloning of Saccharomyces cerevisiae STE5 as a suppressor of a Ste20 protein kinase mutant: structural and functional similarity of Ste5 to Far1.
    Mol Gen Genet. 1993 Nov;241(3-4):241-54 PMID: 8246877
  9. Proteins regulating Ras and its relatives.
    Nature. 1993 Dec 16;366(6456):643-54 PMID: 8259209
  10. A brain serine/threonine protein kinase activated by Cdc42 and Rac1.
    Nature. 1994 Jan 6;367(6458):40-6 PMID: 8107774
  11. Activation of NADPH oxidase involves the dissociation of p21rac from its inhibitory GDP/GTP exchange protein (rhoGDI) followed by its translocation to the plasma membrane.
    Biochem J. 1994 Mar 15;298 Pt 3:585-91 PMID: 8141770
  12. Regulation and function of the Rho subfamily of small GTPases.
    Curr Opin Genet Dev. 1994 Feb;4(1):77-81 PMID: 8193544
  13. Activation of phosphoinositide 3-kinase activity by Cdc42Hs binding to p85.
    J Biol Chem. 1994 Jul 22;269(29):18727-30 PMID: 8034624
  14. Interaction of Rac with p67phox and regulation of phagocytic NADPH oxidase activity.
    Science. 1994 Jul 22;265(5171):531-3 PMID: 8036496
  15. Raf meets Ras: completing the framework of a signal transduction pathway.
    Trends Biochem Sci. 1994 Jul;19(7):279-83 PMID: 8048167
  16. Dbl and Vav mediate transformation via mitogen-activated protein kinase pathways that are distinct from those activated by oncogenic Ras.
    Mol Cell Biol. 1994 Oct;14(10):6848-57 PMID: 7935402
  17. Oncogene ect2 is related to regulators of small GTP-binding proteins.
    Nature. 1993 Apr 1;362(6419):462-5 PMID: 8464478
  18. Peptide mapping and purification of phosphopeptides using high-performance liquid chromatography.
    Methods Enzymol. 1983;99:37-48 PMID: 6316098
  19. Organic extraction of Pi with isobutanol/toluene.
    Anal Biochem. 1984 May 1;138(2):416-20 PMID: 6742419
  20. Complexes of polyoma virus medium T antigen and cellular proteins.
    Proc Natl Acad Sci U S A. 1985 Dec;82(23):7952-4 PMID: 2415976
  21. Separation of peptides dissolved in a sodium dodecyl sulfate solution by reversed-phase liquid chromatography: removal of sodium dodecyl sulfate from peptides using an ion-exchange precolumn.
    Anal Biochem. 1990 May 1;186(2):264-8 PMID: 2363498
  22. 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
  23. Activation of the NADPH oxidase involves the small GTP-binding protein p21rac1.
    Nature. 1991 Oct 17;353(6345):668-70 PMID: 1922386
  24. Catalysis of guanine nucleotide exchange on the CDC42Hs protein by the dbl oncogene product.
    Nature. 1991 Nov 28;354(6351):311-4 PMID: 1956381
  25. Regulation of phagocyte oxygen radical production by the GTP-binding protein Rac 2.
    Science. 1991 Dec 6;254(5037):1512-5 PMID: 1660188
  26. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors.
    Cell. 1992 Aug 7;70(3):389-99 PMID: 1643657
  27. The small GTP-binding protein rac regulates growth factor-induced membrane ruffling.
    Cell. 1992 Aug 7;70(3):401-10 PMID: 1643658
  28. Diversity and versatility of GTPase activating proteins for the p21rho subfamily of ras G proteins detected by a novel overlay assay.
    J Biol Chem. 1992 Aug 15;267(23):16025-8 PMID: 1644791
  29. Association between GTPase activators for Rho and Ras families.
    Nature. 1992 Sep 10;359(6391):153-4 PMID: 1522900
  30. Dominant-negative mutants of a yeast G-protein beta subunit identify two functional regions involved in pheromone signalling.
    EMBO J. 1992 Dec;11(13):4805-13 PMID: 1464310
  31. Involvement of rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI) in cell motility.
    Mol Cell Biol. 1993 Jan;13(1):72-9 PMID: 8417362
  32. 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
  33. Regulation of cytoplasmic division of Xenopus embryo by rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI).
    J Cell Biol. 1993 Mar;120(5):1187-95 PMID: 8436590
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1995-05-01
Pages
1970-8
Language
English
Region
England
NLM ID
8208664
PMCID
PMC398296
Subset
IM
Databases
GENBANK
U25975, U35345
Corrections
ErratumIn
PMID: 7556080
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