RRC ID 65044
Author Yamada H, Tsutsumi K, Nakazawa Y, Shibagaki Y, Hattori S, Ohta Y.
Title Src Family Tyrosine Kinase Signaling Regulates FilGAP through Association with RBM10.
Journal PLoS One
Abstract FilGAP is a Rac-specific GTPase-activating protein (GAP) that suppresses lamellae formation. In this study, we have identified RBM10 (RNA Binding Motif domain protein 10) as a FilGAP-interacting protein. Although RBM10 is mostly localized in the nuclei in human melanoma A7 cells, forced expression of Src family tyrosine kinase Fyn induced translocation of RBM10 from nucleus into cell peripheries where RBM10 and FilGAP are co-localized. The translocation of RBM10 from nucleus appears to require catalytic activity of Fyn since kinase-negative Fyn mutant failed to induce translocation of RBM10 in A7 cells. When human breast carcinoma MDA-MB-231 cells are spreading on collagen-coated coverslips, endogenous FilGAP and RBM10 were localized at the cell periphery with tyrosine-phosphorylated proteins. RBM10 appears to be responsible for targeting FilGAP at the cell periphery because depletion of RBM10 by siRNA abrogated peripheral localization of FilGAP during cell spreading. Association of RBM10 with FilGAP may stimulate RacGAP activity of FilGAP. First, forced expression of RBM10 suppressed FilGAP-mediated cell spreading on collagen. Conversely, depletion of endogenous RBM10 by siRNA abolished FilGAP-mediated suppression of cell spreading on collagen. Second, FilGAP suppressed formation of membrane ruffles induced by Fyn and instead produced spiky cell protrusions at the cell periphery. This protrusive structure was also induced by depletion of Rac, suggesting that the formation of protrusions may be due to suppression of Rac by FilGAP. We found that depletion of RBM10 markedly reduced the formation of protrusions in cells transfected with Fyn and FilGAP. Finally, depletion of RBM10 blocked FilGAP-mediated suppression of ruffle formation induced by EGF. Taken together, these results suggest that Src family tyrosine kinase signaling may regulate FilGAP through association with RBM10.
Volume 11(1)
Pages e0146593
Published 2016-1-1
DOI 10.1371/journal.pone.0146593
PII PONE-D-15-32284
PMID 26751795
PMC PMC4709192
MeSH Breast Neoplasms / metabolism* Catalysis Cell Line, Tumor Cell Nucleus / metabolism Collagen / chemistry DNA, Complementary / metabolism Female GTPase-Activating Proteins / metabolism* Gene Expression Regulation, Neoplastic* HEK293 Cells Humans Microscopy, Fluorescence Plasmids / metabolism RNA, Small Interfering / metabolism RNA-Binding Proteins / metabolism* Signal Transduction* Tyrosine / chemistry src-Family Kinases / metabolism*
IF 2.74
Human and Animal Cells 293(RCB1637)