TY - JOUR
T1 - Wnt5A activates the calpain-mediated cleavage of filamin a
AU - O'Connell, Michael P.
AU - Fiori, Jennifer L.
AU - Baugher, Katherine M.
AU - Indig, Fred E.
AU - French, Amanda D.
AU - Camilli, Tura C.
AU - Frank, Brittany P.
AU - Earley, Rachel
AU - Hoek, Keith S.
AU - Hasskamp, Joanne H.
AU - Elias, E. George
AU - Taub, Dennis D.
AU - Bernier, Michel
AU - Weeraratna, Ashani T.
N1 - Funding Information:
We thank Liqun Jiang for generously providing calpain inhibitor III. This research was supported by the Intramural Research Program (IRP) of the National Institute on Aging (NIA), National Institutes of Health (NIH). K.S.H. was supported by the Swiss National Foundation (3100A0-103671) and Oncosuisse (OCS-01927-08-2006).
PY - 2009/7
Y1 - 2009/7
N2 - We have previously shown that Wnt5A and ROR2, an orphan tyrosine kinase receptor, interact to mediate melanoma cell motility. In other cell types, this can occur through the interaction of ROR2 with the cytoskeletal protein filamin A. Here, we found that filamin A protein levels correlated with Wnt5A levels in melanoma cells. Small interfering RNA (siRNA) knockdown of WNT5A decreased filamin A expression. Knockdown of filamin A also corresponded to a decrease in melanoma cell motility. In metastatic cells, filamin A expression was predominant in the cytoplasm, which western analysis indicated was due to the cleavage of filamin A in these cells. Treatment of nonmetastatic melanoma cells with recombinant Wnt5A increased filamin A cleavage, and this could be prevented by the knockdown of ROR2 expression. Further, BAPTA-AM chelation of intracellular calcium also inhibited filamin A cleavage, leading to the hypothesis that Wnt5A/ROR2 signaling could cleave filamin A through activation of calcium-activated proteases, such as calpains. Indeed, WNT5A knockdown decreased calpain 1 expression, and by inhibiting calpain 1 either pharmacologically or using siRNA, it decreased cell motility. Our results indicate that Wnt5A activates calpain-1, leading to the cleavage of filamin A, which results in a remodeling of the cytoskeleton and an increase in melanoma cell motility.
AB - We have previously shown that Wnt5A and ROR2, an orphan tyrosine kinase receptor, interact to mediate melanoma cell motility. In other cell types, this can occur through the interaction of ROR2 with the cytoskeletal protein filamin A. Here, we found that filamin A protein levels correlated with Wnt5A levels in melanoma cells. Small interfering RNA (siRNA) knockdown of WNT5A decreased filamin A expression. Knockdown of filamin A also corresponded to a decrease in melanoma cell motility. In metastatic cells, filamin A expression was predominant in the cytoplasm, which western analysis indicated was due to the cleavage of filamin A in these cells. Treatment of nonmetastatic melanoma cells with recombinant Wnt5A increased filamin A cleavage, and this could be prevented by the knockdown of ROR2 expression. Further, BAPTA-AM chelation of intracellular calcium also inhibited filamin A cleavage, leading to the hypothesis that Wnt5A/ROR2 signaling could cleave filamin A through activation of calcium-activated proteases, such as calpains. Indeed, WNT5A knockdown decreased calpain 1 expression, and by inhibiting calpain 1 either pharmacologically or using siRNA, it decreased cell motility. Our results indicate that Wnt5A activates calpain-1, leading to the cleavage of filamin A, which results in a remodeling of the cytoskeleton and an increase in melanoma cell motility.
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U2 - 10.1038/jid.2008.433
DO - 10.1038/jid.2008.433
M3 - Article
C2 - 19177143
AN - SCOPUS:67449128692
SN - 0022-202X
VL - 129
SP - 1782
EP - 1789
JO - Journal of Investigative Dermatology
JF - Journal of Investigative Dermatology
IS - 7
ER -