![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
Vol. 15, Issue 3, 1003-1010, March 2004
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Department of Tumor Virology, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan
Submitted August 20, 2003;
Revised October 28, 2003;
Accepted November 18, 2003
Monitoring Editor: Anne Ridley
A major function of Rho-family GTPases is to regulate the organization of the actin cytoskeleton; filopodia, lamellipodia, and stress fiber are regarded as typical phenotypes of the activated Cdc42, Rac, and Rho, respectively. Using probes based on fluorescent resonance energy transfer, we report on the spatiotemporal regulation of Rac1 and Cdc42 at lamellipodia and membrane ruffles. In epidermal growth factor (EGF)-stimulated Cos1 and A431 cells, both Rac1 and Cdc42 were activated diffusely at the plasma membrane, followed by lamellipodial protrusion and membrane ruffling. Although Rac1 activity subsided rapidly, Cdc42 activity was sustained at lamellipodia. A critical role of Cdc42 in these EGF-induced morphological changes was demonstrated as follows. First, phorbol 12-myristate 13-acetate, which activated Rac1 but not Cdc42, could not induce full-grown lamellipodia in Cos1 cells. Second, a GTPase-activating protein for Cdc42, KIAA1204/CdGAP, inhibited lamellipodial protrusion and membrane ruffling without interfering with Rac1 activation. Third, expression of the Cdc42-binding domain of N-WASP inhibited the EGF-induced morphological changes. Therefore, Rac1 and Cdc42 seem to synergistically induce lamellipodia and membrane ruffles in EGF-stimulated Cos1 cells and A431 cells.
Abbreviations used: CFP, cyan fluorescent protein; EGF, epidermal growth factor; FRET, fluorescent resonance energy transfer; GAP, GTPase-activating protein; GDI, guanine nucleotide dissociation inhibitor; GEF, guanine nucleotide exchange factor; GFP, green fluorescent protein; mRFP, monomeric red fluorescent protein; PMA, phorbol 12-myristate 13-acetate; TPEM, two-photon excitation fluorescence microscopy; YFP, yellow fluorescent protein.
Online version of this article contains supplementary videos for some figures. Online version is available at www.molbiolcell.org.
* Corresponding author. E-mail address: matsudam{at}biken.osakau.ac.jp.
This article has been cited by other articles:
|
|
 |
|
  K. Makrogianneli, L. M. Carlin, M. D. Keppler, D. R. Matthews, E. Ofo, A. Coolen, S. M. Ameer-Beg, P. R. Barber, B. Vojnovic, and T. Ng Integrating Receptor Signal Inputs That Influence Small Rho GTPase Activation Dynamics at the Immunological Synapse Mol. Cell. Biol., June 1, 2009; 29(11): 2997 - 3006. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P.R Barber, S.M Ameer-Beg, J Gilbey, L.M Carlin, M Keppler, T.C Ng, and B Vojnovic Multiphoton time-domain fluorescence lifetime imaging microscopy: practical application to protein-protein interactions using global analysis J R Soc Interface, February 6, 2009; 6(Suppl_1): S93 - S105. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. E. Itoh, E. Kiyokawa, K. Aoki, T. Nishioka, T. Akiyama, and M. Matsuda Phosphorylation and activation of the Rac1 and Cdc42 GEF Asef in A431 cells stimulated by EGF J. Cell Sci., August 15, 2008; 121(16): 2635 - 2642. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Liu, D. Li, B. Cao, Y.-x. Li, R. Herva, Y.-s. Piao, and Y.-l. Wang Expression and Localization of SWAP-70 in Human Fetomaternal Interface and Placenta During Tubal Pregnancy and Normal Placentation J. Histochem. Cytochem., July 1, 2007; 55(7): 701 - 708. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Chen, T. D. Abair, M. R. Ibanez, Y. Su, M. R. Frey, R. S. Dise, D. B. Polk, A. B. Singh, R. C. Harris, R. Zent, et al. Integrin {alpha}1{beta}1 Controls Reactive Oxygen Species Synthesis by Negatively Regulating Epidermal Growth Factor Receptor-Mediated Rac Activation Mol. Cell. Biol., May 1, 2007; 27(9): 3313 - 3326. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Takaya, T. Kamio, M. Masuda, N. Mochizuki, H. Sawa, M. Sato, K. Nagashima, A. Mizutani, A. Matsuno, E. Kiyokawa, et al. R-Ras Regulates Exocytosis by Rgl2/Rlf-mediated Activation of RalA on Endosomes Mol. Biol. Cell, May 1, 2007; 18(5): 1850 - 1860. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. E. Goldfinger, C. Ptak, E. D. Jeffery, J. Shabanowitz, D. F. Hunt, and M. H. Ginsberg RLIP76 (RalBP1) is an R-Ras effector that mediates adhesion-dependent Rac activation and cell migration J. Cell Biol., September 11, 2006; 174(6): 877 - 888. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. Atienza, N. Yu, X. Wang, X. Xu, and Y. Abassi Label-Free and Real-Time Cell-Based Kinase Assay for Screening Selective and Potent Receptor Tyrosine Kinase Inhibitors Using Microelectronic Sensor Array J Biomol Screen, September 1, 2006; 11(6): 634 - 643. [Abstract] [PDF]
|
|
|
|
|
|
L. Za, C. Albertinazzi, S. Paris, M. Gagliani, C. Tacchetti, and I. de Curtis {beta}PIX controls cell motility and neurite extension by regulating the distribution of GIT1 J. Cell Sci., July 1, 2006; 119(13): 2654 - 2666. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. G. Lee and E. P. Kay Cross-talk among Rho GTPases Acting Downstream of PI 3-Kinase Induces Mesenchymal Transformation of Corneal Endothelial Cells Mediated by FGF-2. Invest. Ophthalmol. Vis. Sci., June 1, 2006; 47(6): 2358 - 2368. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Cai, N. Holoweckyj, M. D. Schaller, and J. E. Bear Phosphorylation of Coronin 1B by Protein Kinase C Regulates Interaction with Arp2/3 and Cell Motility J. Biol. Chem., September 9, 2005; 280(36): 31913 - 31923. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Kurokawa and M. Matsuda Localized RhoA Activation as a Requirement for the Induction of Membrane Ruffling Mol. Biol. Cell, September 1, 2005; 16(9): 4294 - 4303. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Parsons, J. Monypenny, S. M. Ameer-Beg, T. H. Millard, L. M. Machesky, M. Peter, M. D. Keppler, G. Schiavo, R. Watson, J. Chernoff, et al. Spatially Distinct Binding of Cdc42 to PAK1 and N-WASP in Breast Carcinoma Cells Mol. Cell. Biol., March 1, 2005; 25(5): 1680 - 1695. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Takaya, Y. Ohba, K. Kurokawa, and M. Matsuda RalA Activation at Nascent Lamellipodia of Epidermal Growth Factor-stimulated Cos7 Cells and Migrating Madin-Darby Canine Kidney Cells Mol. Biol. Cell, June 1, 2004; 15(6): 2549 - 2557. [Abstract] [Full Text] [PDF]
|
|
