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© The Rockefeller University Press, 0021-9525/1999//1009 $5.00
The Journal of Cell Biology, Volume 147, Number 5, , 1999 1009-1022

Rac Downregulates Rho Activity

Eva E. Sander^a, Jean P. ten Klooster^a, Sanne van Delft^a, Rob A. van der Kammen^a, and John G. Collard^a

^a The Netherlands Cancer Institute, Division of Cell Biology, 1066 CX Amsterdam, The Netherlands
The Netherlands Cancer Institute, Division of Cell Biology, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.31-20-512-194431-20-512-1932

jcoll{at}nki.nl

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.

Key Words: cell–cell adhesion • migration • Rho-like GTPases • Tiam1

© 1999 The Rockefeller University Press

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• Brumby, A., Secombe, J., Horsfield, J., Coombe, M., Amin, N., Coates, D., Saint, R., Richardson, H. (2004). A Genetic Screen for Dominant Modifiers of a cyclin E Hypomorphic Mutation Identifies Novel Regulators of S-Phase Entry in Drosophila. Genetics 168: 227-251 [Abstract] [Full Text]  
• Liang, X., Draghi, N. A., Resh, M. D. (2004). Signaling from Integrins to Fyn to Rho Family GTPases Regulates Morphologic Differentiation of Oligodendrocytes. J. Neurosci. 24: 7140-7149 [Abstract] [Full Text]  
• Turner, L. J., Nicholls, S., Hall, A. (2004). The Activity of the Plexin-A1 Receptor Is Regulated by Rac. J. Biol. Chem. 279: 33199-33205 [Abstract] [Full Text]  
• Malliri, A., van Es, S., Huveneers, S., Collard, J. G. (2004). The Rac Exchange Factor Tiam1 Is Required for the Establishment and Maintenance of Cadherin-based Adhesions. J. Biol. Chem. 279: 30092-30098 [Abstract] [Full Text]  
• Schotte, P., Denecker, G., Van Den Broeke, A., Vandenabeele, P., Cornelis, G. R., Beyaert, R. (2004). Targeting Rac1 by the Yersinia Effector Protein YopE Inhibits Caspase-1-mediated Maturation and Release of Interleukin-1{beta}. J. Biol. Chem. 279: 25134-25142 [Abstract] [Full Text]  
• Zimmerman, A. W., Nelissen, J. M. D. T., van Emst-de Vries, S. E., Willems, P. H. G. M., de Lange, F., Collard, J. G., van Leeuwen, F. N., Figdor, C. G. (2004). Cytoskeletal restraints regulate homotypic ALCAM-mediated adhesion through PKC{alpha} independently of Rho-like GTPases. J. Cell Sci. 117: 2841-2852 [Abstract] [Full Text]  
• Barker, T. H., Pallero, M. A., MacEwen, M. W., Tilden, S. G., Woods, A., Murphy-Ullrich, J. E., Hagood, J. S. (2004). Thrombospondin-1-induced Focal Adhesion Disassembly in Fibroblasts Requires Thy-1 Surface Expression, Lipid Raft Integrity, and Src Activation. J. Biol. Chem. 279: 23510-23516 [Abstract] [Full Text]  
• Bailly, K., Ridley, A. J., Hall, S. M., Haworth, S. G. (2004). RhoA Activation by Hypoxia in Pulmonary Arterial Smooth Muscle Cells Is Age and Site Specific. Circ. Res. 94: 1383-1391 [Abstract] [Full Text]  

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