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Eur J Cell Biol. 2002 Jun;81(6):351-62.

Microtubule dynamics differentially regulates Rho and Rac activity and triggers Rho-independent stress fiber formation in macrophage polykaryons.

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  • 1Laboratoire de Biologie MolĂ©culaire et Cellulaire, Ecole Normale SupĂ©rieure de Lyon UMR5665, CNRS, ENS Equipe INRA, France.

Abstract

Multinucleated giant cells (MNGC) derived from avian peripheral blood monocytes present a dense microtubular network emanating from peripherally located centrosomes. We were interested to study how microtubule and F-actin cytoskeletons cooperate in MNGC to maintain cell shape. Microtubule depolymerization by nocodazole triggered the reorganization of the F-actin cytoskeleton in MNGC that is normally organized into podosomes, cortical actin filaments and membrane ruffles. After nocodazole treatment, F-actin was redistributed into unusual transverse fibers associated with focal adhesion plaques. When microtubules were allowed to repolymerize after nocodazole removal, F-actin appeared transiently, together with the small GTPase Rac, in large membrane ruffles. Using affinity precipitation assays, we show that microtubule depolymerization leads to activation of Rho and inhibition of Rac, whereas microtubule repolymerization induces Rac activation and Rho inhibition. Thus, the level of microtubule polymerization inversely regulates Rho and Rac activity in MNGC. Moreover, using C3 exoenzyme, a known inhibitor of Rho, we demonstrate that both the F-actin fiber formation in response to microtubule depolymerization and the formation of membrane ruffles after microtubule repolymerization occur in C3-treated MNGC, indicating that Rho is not required for these events.

PMID:
12113476
DOI:
10.1078/0171-9335-00255
[PubMed - indexed for MEDLINE]
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