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MBC in Press, published online ahead of print September 5, 2003
Mol. Biol. Cell 10.1091/mbc.E02-11-0739

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Submitted on November 18, 2002
Revised on June 26, 2003
Accepted on July 14, 2003

Rapid Microtubule-dependent Induction of Neurite-like Extensions in NIH 3T3 Fibroblasts by Inhibition of ROCK and Cbl

Robin M. Scaife1*, Didier Job2, and Wallace Y. Langdon3

1 Department of Pathology, University of Western Australia, Crawley, WA 6009, Australia and DBMS-CEA, Grenoble, France
2 DBMS-CEA, Grenoble, France
3 Department of Pathology, University of Western Australia, Crawley, WA 6009, Australia

* Corresponding author. E-mail address: rscaife{at}cyllene.uwa.edu.au.

A number of key cellular functions, such as morphological differentiation and cell motility, are closely associated with changes in cytoskeletal dynamics. Many of the principal signaling components involved in actin cytoskeletal dynamics have been identified, and these have been shown to be critically involved in cell motility. By contrast, signaling to microtubules remains relatively uncharacterised and the importance of signaling pathways in modulation of microtubule dynamics has so far not been established clearly. We report here that the Rho-effector ROCK and the multi-adaptor proto-oncoprotein Cbl can profoundly affect the microtubule cytoskeleton. Simultaneous inhibition of these two signaling molecules induces a dramatic rearrangement of the microtubule cytoskeleton into microtubule bundles. The formation of these microtubule bundles, which does not involve signaling by Rac, Cdc42, Crk, PI 3-kinase and Abl, is sufficient to induce distinct neurite-like extensions in NIH 3T3 fibroblasts, even in the absence of microfilaments. This novel microtubule-dependent function that promotes neurite-like extensions is not dependent on net changes in microtubule polymerization or stabilization, but rather involves selective elongation and reorganisation of microtubules into long bundles.




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