Wnt-3a-dependent cell motility involves RhoA activation and is specifically regulated by dishevelled-2.
Wnts stimulate cell migration, although the mechanisms responsible for this effect are not fully understood. To investigate the pathways that mediate Wnt-dependent cell motility, we treated Chinese hamster ovary cells with Wnt-3a-conditioned medium and monitored changes in cell shape and movement. Wnt-3a induced cell spreading, formation of protrusive structures, reorganization of stress fibers and migration. Although Wnt-3a stabilized beta-catenin, two inhibitors of the beta-catenin/canonical pathway, Dickkopf-1 and a dominant-negative T cell factor construct, did not reduce motility. The small GTPase RhoA also was activated by Wnt-3a. In contrast to beta-catenin signaling, inhibition of Rho kinase partially blocked motility. Because Dishevelled (Dvl) proteins are effectors of both canonical and noncanonical Wnt signaling, we used immunofluorescent analysis and small interference RNA technology to evaluate the role of Dvl in cell motility. Specific knock-down of Dvl-2 expression markedly reduced Wnt-3a-dependent changes in cell shape and movement, suggesting that this Dvl isoform had a predominant role in mediating Wnt-3a-dependent motility in Chinese hamster ovary cells.
Endo
Yoshimi
Y
Laboratories of Cellular and Molecular Biology, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA.
Wolf
Vladimir
V
Muraiso
Kanae
K
Kamijo
Keiju
K
Soon
Lilian
L
Uren
Aykut
A
Barshishat-Küpper
Michal
M
Rubin
Jeffrey S
JS
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D016466:CHO Cells; D048430:Cell Shape / Q000187:drug effects*; D002633:Chemotaxis / Q000187:drug effects*; D006224:Cricetinae; D003412:Cricetulus; D017077:Culture Media, Conditioned / Q000494:pharmacology; D000072261:Dishevelled Proteins; D004789:Enzyme Activation / Q000187:drug effects; D051379:Mice; D010750:Phosphoproteins; D011506:Proteins / Q000378:metabolism / Q000494:pharmacology*; D015398:Signal Transduction; D051153:Wnt Proteins; D060508:Wnt3 Protein; D060509:Wnt3A Protein; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
D048868:Adaptor Proteins, Signal Transducing; D017077:Culture Media, Conditioned; D000072261:Dishevelled Proteins; C000606582:Dvl2 protein, mouse; D010750:Phosphoproteins; D011506:Proteins; D051153:Wnt Proteins; D060508:Wnt3 Protein; D060509:Wnt3A Protein; C555511:Wnt3a protein, mouse; D020742:rhoA GTP-Binding Protein