Home LiteratureArticle Details
PMID: 10579721 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Rac downregulates Rho activity: reciprocal balance between both GTPases determines cellular morphology and migratory behavior.

The Journal of cell biology ·Vol. 147 ·No. 5 ·1999-11-29 ·Pages 1009-22

Sander EE, ten Klooster JP, van Delft S, van der Kammen RA, Collard JG

Abstract

Using biochemical assays to determine the activation state of Rho-like GTPases, we show that the guanine nucleotide exchange factor Tiam1 functions as a specific activator of Rac but not Cdc42 or Rho in NIH3T3 fibroblasts. Activation of Rac by Tiam1 induces an epithelial-like morphology with functional cadherin-based adhesions and inhibits migration of fibroblasts. This epithelial phenotype is characterized by Rac-mediated effects on Rho activity. Transient PDGF-induced as well as sustained Rac activation by Tiam1 or V12Rac downregulate Rho activity. We found that Cdc42 also downregulates Rho activity. Neither V14Rho or N19Rho affects Rac activity, suggesting unidirectional signaling from Rac towards Rho. Downregulation of Rho activity occurs independently of Rac- induced cytoskeletal changes and cell spreading. Moreover, Rac effector mutants that are defective in mediating cytoskeleton changes or Jun kinase activation both downregulate Rho activity, suggesting that neither of these Rac signaling pathways are involved in the regulation of Rho. Restoration of Rho activity in Tiam1-expressing cells by expression of V14Rho results in reversion of the epithelioid phenotype towards a migratory, fibroblastoid morphology. We conclude that Rac signaling is able to antagonize Rho activity directly at the GTPase level, and that the reciprocal balance between Rac and Rho activity determines cellular morphology and migratory behavior in NIH3T3 fibroblasts.

MeSH Terms
3T3 Cells Animals Cadherins/metabolism Cell Adhesion/physiology Cell Line Cell Movement/physiology Cytoskeleton/metabolism Down-Regulation/physiology Epithelial Cells/cytology,metabolism GTP Phosphohydrolases/metabolism GTPase-Activating Proteins/metabolism Guanine Nucleotide Exchange Factors Mice Phenotype Platelet-Derived Growth Factor/metabolism Protein Biosynthesis Proteins Signal Transduction/physiology T-Lymphoma Invasion and Metastasis-inducing Protein 1 cdc42 GTP-Binding Protein/metabolism rac GTP-Binding Proteins/metabolism
Chemicals
Cadherins GTPase-Activating Proteins Guanine Nucleotide Exchange Factors Platelet-Derived Growth Factor Proteins T-Lymphoma Invasion and Metastasis-inducing Protein 1 Tiam1 protein, mouse GTP Phosphohydrolases cdc42 GTP-Binding Protein rac GTP-Binding Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Sander E E
The Netherlands Cancer Institute, Division of Cell Biology, 1066 CX Amsterdam, The Netherlands.
ten Klooster J P
van Delft S
van der Kammen R A
Collard J G
References (51)
51 references, click to expand
  1. Oncogenic activity of Tiam1 and Rac1 in NIH3T3 cells.
    Oncogene. 1995 Dec 7;11(11):2215-21 PMID: 8570171
  2. Interplay between Rac and Rho in the control of substrate contact dynamics.
    Curr Biol. 1999 Jun 17;9(12):640-8 PMID: 10375527
  3. The novel catenin p120cas binds classical cadherins and induces an unusual morphological phenotype in NIH3T3 fibroblasts.
    Exp Cell Res. 1996 Jun 15;225(2):328-37 PMID: 8660921
  4. Rhotekin, a new putative target for Rho bearing homology to a serine/threonine kinase, PKN, and rhophilin in the rho-binding domain.
    J Biol Chem. 1996 Jun 7;271(23):13556-60 PMID: 8662891
  5. Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase)
    Science. 1996 Jul 12;273(5272):245-8 PMID: 8662509
  6. The N-terminal pleckstrin, coiled-coil, and IQ domains of the exchange factor Ras-GRF act cooperatively to facilitate activation by calcium.
    Mol Cell Biol. 1996 Sep;16(9):4888-96 PMID: 8756648
  7. Still life, a protein in synaptic terminals of Drosophila homologous to GDP-GTP exchangers.
    Science. 1997 Jan 24;275(5299):543-7 PMID: 8999801
  8. Rho family GTPases and neuronal growth cone remodelling: relationship between increased complexity induced by Cdc42Hs, Rac1, and acetylcholine and collapse induced by RhoA and lysophosphatidic acid.
    Mol Cell Biol. 1997 Mar;17(3):1201-11 PMID: 9032247
  9. Identification and characterization of hPEM-2, a guanine nucleotide exchange factor specific for Cdc42.
    J Biol Chem. 1999 Nov 19;274(47):33587-93 PMID: 10559246
  10. Rac regulates phosphorylation of the myosin-II heavy chain, actinomyosin disassembly and cell spreading.
    Nat Cell Biol. 1999 Aug;1(4):242-8 PMID: 10559923
  11. Molecular cloning of cDNAs encoding a guanine-nucleotide-releasing factor for Ras p21.
    Nature. 1992 Jul 23;358(6384):351-4 PMID: 1379346
  12. 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
  13. The small GTP-binding protein rac regulates growth factor-induced membrane ruffling.
    Cell. 1992 Aug 7;70(3):401-10 PMID: 1643658
  14. Identification of an invasion-inducing gene, Tiam-1, that encodes a protein with homology to GDP-GTP exchangers for Rho-like proteins.
    Cell. 1994 May 20;77(4):537-49 PMID: 7999144
  15. Signal transduction pathways regulating Rho-mediated stress fibre formation: requirement for a tyrosine kinase.
    EMBO J. 1994 Jun 1;13(11):2600-10 PMID: 7516876
  16. Inhibition of lysophosphatidate- and thrombin-induced neurite retraction and neuronal cell rounding by ADP ribosylation of the small GTP-binding protein Rho.
    J Cell Biol. 1994 Aug;126(3):801-10 PMID: 8045941
  17. Isolation and characterization of the faciogenital dysplasia (Aarskog-Scott syndrome) gene: a putative Rho/Rac guanine nucleotide exchange factor.
    Cell. 1994 Nov 18;79(4):669-78 PMID: 7954831
  18. Regulation of scatter factor/hepatocyte growth factor responses by Ras, Rac, and Rho in MDCK cells.
    Mol Cell Biol. 1995 Feb;15(2):1110-22 PMID: 7823927
  19. 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
  20. 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
  21. A role for Rac in Tiam1-induced membrane ruffling and invasion.
    Nature. 1995 May 25;375(6529):338-40 PMID: 7753201
  22. c-Src regulates the simultaneous rearrangement of actin cytoskeleton, p190RhoGAP, and p120RasGAP following epidermal growth factor stimulation.
    J Cell Biol. 1995 Jul;130(2):355-68 PMID: 7542246
  23. Phosphorylation and activation of myosin by Rho-associated kinase (Rho-kinase).
    J Biol Chem. 1996 Aug 23;271(34):20246-9 PMID: 8702756
  24. Episomal vectors rapidly and stably produce high-titer recombinant retrovirus.
    Hum Gene Ther. 1996 Aug 1;7(12):1405-13 PMID: 8844199
  25. Rac and Cdc42 induce actin polymerization and G1 cell cycle progression independently of p65PAK and the JNK/SAPK MAP kinase cascade.
    Cell. 1996 Nov 1;87(3):519-29 PMID: 8898204
  26. RAC regulation of actin polymerization and proliferation by a pathway distinct from Jun kinase.
    Science. 1996 Nov 22;274(5291):1374-6 PMID: 8910277
  27. Human p21-activated kinase (Pak1) regulates actin organization in mammalian cells.
    Curr Biol. 1997 Mar 1;7(3):202-10 PMID: 9395435
  28. Regulated membrane localization of Tiam1, mediated by the NH2-terminal pleckstrin homology domain, is required for Rac-dependent membrane ruffling and C-Jun NH2-terminal kinase activation.
    J Cell Biol. 1997 Apr 21;137(2):387-98 PMID: 9128250
  29. The small GTPases Rho and Rac are required for the establishment of cadherin-dependent cell-cell contacts.
    J Cell Biol. 1997 Jun 16;137(6):1421-31 PMID: 9182672
  30. Rho GTPases and signaling networks.
    Genes Dev. 1997 Sep 15;11(18):2295-322 PMID: 9308960
  31. The guanine nucleotide exchange factor Tiam1 affects neuronal morphology; opposing roles for the small GTPases Rac and Rho.
    J Cell Biol. 1997 Nov 3;139(3):797-807 PMID: 9348295
  32. Targeting of Tiam1 to the plasma membrane requires the cooperative function of the N-terminal pleckstrin homology domain and an adjacent protein interaction domain.
    J Biol Chem. 1997 Nov 7;272(45):28447-54 PMID: 9353304
  33. Rho-stimulated contractility contributes to the fibroblastic phenotype of Ras-transformed epithelial cells.
    Mol Biol Cell. 1997 Nov;8(11):2329-44 PMID: 9362072
  34. Regulation of cell-cell adhesion by rac and rho small G proteins in MDCK cells.
    J Cell Biol. 1997 Nov 17;139(4):1047-59 PMID: 9362522
  35. A novel Cdc42Hs mutant induces cellular transformation.
    Curr Biol. 1997 Oct 1;7(10):794-7 PMID: 9368762
  36. Inhibition of invasion of epithelial cells by Tiam1-Rac signaling.
    Science. 1997 Nov 21;278(5342):1464-6 PMID: 9367959
  37. Platelet-derived growth factor and fibronectin-stimulated migration are differentially regulated by the Rac and extracellular signal-regulated kinase pathways.
    J Biol Chem. 1997 Dec 5;272(49):30688-92 PMID: 9388204
  38. Regulation of cell-cell adhesion of MDCK cells by Cdc42 and Rac1 small GTPases.
    Biochem Biophys Res Commun. 1997 Nov 17;240(2):430-5 PMID: 9388496
  39. Cdc42 and Rac1 induce integrin-mediated cell motility and invasiveness through PI(3)K.
    Nature. 1997 Dec 11;390(6660):632-6 PMID: 9403696
  40. Activation of beta1 integrin signaling stimulates tyrosine phosphorylation of p190RhoGAP and membrane-protrusive activities at invadopodia.
    J Biol Chem. 1998 Jan 2;273(1):9-12 PMID: 9417037
  41. Activation of phosphoinositide 3-OH kinase by the alpha6beta4 integrin promotes carcinoma invasion.
    Cell. 1997 Dec 26;91(7):949-60 PMID: 9428518
  42. Rho GTPases and the actin cytoskeleton.
    Science. 1998 Jan 23;279(5350):509-14 PMID: 9438836
  43. Rac regulates integrin-mediated spreading and increased adhesion of T lymphocytes.
    Mol Cell Biol. 1998 Jul;18(7):3936-46 PMID: 9632778
  44. Activation of Rac and Cdc42 by integrins mediates cell spreading.
    Mol Biol Cell. 1998 Jul;9(7):1863-71 PMID: 9658176
  45. Invasion of T-lymphoma cells: cooperation between Rho family GTPases and lysophospholipid receptor signaling.
    EMBO J. 1998 Jul 15;17(14):4066-74 PMID: 9670021
  46. Matrix-dependent Tiam1/Rac signaling in epithelial cells promotes either cell-cell adhesion or cell migration and is regulated by phosphatidylinositol 3-kinase.
    J Cell Biol. 1998 Nov 30;143(5):1385-98 PMID: 9832565
  47. Regulation of the small GTP-binding protein Rho by cell adhesion and the cytoskeleton.
    EMBO J. 1999 Feb 1;18(3):578-85 PMID: 9927417
  48. Inhibition of myosin light chain kinase by p21-activated kinase.
    Science. 1999 Mar 26;283(5410):2083-5 PMID: 10092231
  49. p21-activated kinase 1 (Pak1) regulates cell motility in mammalian fibroblasts.
    J Cell Biol. 1999 May 17;145(4):837-49 PMID: 10330410
  50. Identification of the stef gene that encodes a novel guanine nucleotide exchange factor specific for Rac1.
    J Biol Chem. 1999 Jun 18;274(25):17837-44 PMID: 10364228
  51. The pleckstrin homology domain: an intriguing multifunctional protein module.
    Bioessays. 1996 Jan;18(1):35-46 PMID: 8593162
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.147.5.1009
Published
1999-11-29
Pages
1009-22
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2169355
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.147.5.1009 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