Cdc42Hs and Rac1 GTPases induce the collapse of the vimentin intermediate filament network.
In this study we show that expression of active Cdc42Hs and Rac1 GTPases, two Rho family members, leads to the reorganization of the vimentin intermediate filament (IF) network, showing a perinuclear collapse. Cdc42Hs displays a stronger effect than Rac1 as 90% versus 75% of GTPase-expressing cells show vimentin collapse. Similar vimentin IF modifications were observed when endogenous Cdc42Hs was activated by bradykinin treatment, endogenous Rac1 by platelet-derived growth factor/epidermal growth factor, or both endogenous proteins upon expression of active RhoG. This reorganization of the vimentin IF network is not associated with any significant increase in soluble vimentin. Using effector loop mutants of Cdc42Hs and Rac1, we show that the vimentin collapse is mostly independent of CRIB (Cdc42Hs or Rac-interacting binding)-mediated pathways such as JNK or PAK activation but is associated with actin reorganization. This does not result from F-actin depolymerization, because cytochalasin D treatment or Scar-WA expression have merely no effect on vimentin organization. Finally, we show that genistein treatment of Cdc42 and Rac1-expressing cells strongly reduces vimentin collapse, whereas staurosporin, wortmannin, LY-294002, R(p)-cAMP, or RII, the regulatory subunit of protein kinase A, remain ineffective. Moreover, we detected an increase in cellular tyrosine phosphorylation content after Cdc42Hs and Rac1 expression without modification of the vimentin phosphorylation status. These data indicate that Cdc42Hs and Rac1 GTPases control vimentin IF organization involving tyrosine phosphorylation events.
Meriane
M
M
Centre de Recherche de Biochimie Macromoléculaire, CNRS, UPR 1086, 1919 Route de Mende, Montpellier 34293, Cedex, France.
Mary
S
S
Comunale
F
F
Vignal
E
E
Fort
P
P
Gauthier-Rouviére
C
C
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D004622:Embryo, Mammalian; D005347:Fibroblasts / Q000502:physiology; D006801:Humans; D007382:Intermediate Filaments / Q000502:physiology* / Q000648:ultrastructure; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D014746:Vimentin / Q000378:metabolism*; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism*; D020830:rac1 GTP-Binding Protein / Q000378:metabolism*
D000199:Actins; D011994:Recombinant Proteins; D014746:Vimentin; D019000:Phosphotyrosine; D020764:cdc42 GTP-Binding Protein; D020830:rac1 GTP-Binding Protein