r-ras regulates migration through an interaction with filamin a in melanoma cellsr-ras调节迁移在黑色素瘤细胞通过与细丝蛋白的交互.pdfVIP

R-Ras Regulates Migration through an Interaction with Filamin A in Melanoma Cells 1 2 3 1 2 Joanna E. Gawecka , Genevieve S. Griffiths , Barbro Ek-Rylander , Joe W. Ramos , Michelle L. Matter * 1 Natural Products and Cancer Biology, Cancer Research Center of Hawaii, University of Hawaii at Manoa, Honolulu, Hawaii, United States of America, 2 Department of Cell and Molecular Biology, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, Hawaii, United States of America, 3 Division of Pathology, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital, Huddinge, Sweden Abstract Background: Changes in cell adhesion and migration in the tumor microenvironment are key in the initiation and progression of metastasis. R-Ras is one of several small GTPases that regulate cell adhesion and migration on the extracellular matrix, however the mechanism has not been completely elucidated. Using a yeast two-hybrid approach we sought to identify novel R-Ras binding proteins that might mediate its effects on integrins. Methods and Findings: We identified Filamin A (FLNa) as a candidate interacting protein. FLNa is an actin-binding scaffold protein that also binds to integrin b1, b2 and b7 tails and is associated with diverse cell processes including cell migration. Indeed, M2 melanoma cells require FLNa for motility. We further show that R-Ras and FLNa interact in co- immunoprecipitations and pull-down assays. Deletion of FLNa repeat 3 (FLNaD3) abrogated this interaction. In M2 melanoma cells active R-Ras co-localized with FLNa but did not c