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Published 31 January 2005. doi:10.1083/jcb.200411109
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 168, Number 3, 429-439
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Article

 Concentric zones of active RhoA and Cdc42 around single cell wounds

Hélène A. Benink1 and William M. Bement1,2

1 Department of Zoology, University of Wisconsin-Madison, Madison, WI 53706
2 Program in Cellular and Molecular Biology, University of Wisconsin-Madison, Madison, WI 53706

Correspondence to William M. Bement: wmbement{at}wisc.edu

Rho GTPases control many cytoskeleton-dependent processes, but how they regulate spatially distinct features of cytoskeletal function within a single cell is poorly understood. Here, we studied active RhoA and Cdc42 in wounded Xenopus oocytes, which assemble and close a dynamic ring of actin filaments (F-actin) and myosin-2 around wound sites. RhoA and Cdc42 are rapidly activated around wound sites in a calcium-dependent manner and segregate into distinct, concentric zones around the wound, with active Cdc42 in the approximate middle of the F-actin array and active RhoA on the interior of the array. These zones form before F-actin accumulation, and then move in concert with the closing array. Microtubules and F-actin are required for normal zone organization and dynamics, as is crosstalk between RhoA and Cdc42. Each of the zones makes distinct contributions to the organization and function of the actomyosin wound array. We propose that similar rho activity zones control related processes such as cytokinesis.

Abbreviations used in this paper: 4D, time-lapse multiple focal plane; CARho, constitutively active RhoA; CACdc42, constitutively active Cdc42; DNCdc42, dominant negative Cdc42; MLC, myosin-2 regulatory light chain; mRFP, monomeric red fluorescent protein; ON, overnight; PM, plasma membrane; P-MLC, activated MLC; rGBD, the RhoA-binding domain of rhotekin; wGBD, the Cdc42-binding domain of N-WASP.


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