human mena associates with rac1 small gtpase in glioblastoma cell lines人类mena associates rac1小gtpase胶质母细胞瘤细胞系.pdfVIP

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human mena associates with rac1 small gtpase in glioblastoma cell lines人类mena associates rac1小gtpase胶质母细胞瘤细胞系.pdf

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human mena associates with rac1 small gtpase in glioblastoma cell lines人类mena associates rac1小gtpase胶质母细胞瘤细胞系

Human Mena Associates with Rac1 Small GTPase in Glioblastoma Cell Lines 1. 1. 1 1 1 Morihiro Higashi , Chieko Ishikawa , Jianyong Yu , Akihiro Toyoda , Hidetada Kawana , Kazuo 2 3 1 1 Kurokawa , Michiyuki Matsuda , Motoo Kitagawa , Kenichi Harigaya * 1 Molecular and Tumor Pathology, Chiba University Graduate School of Medicine, Chuo-ku, Chiba, Japan, 2 Molecular Membrane Biology Laboratory, RIKEN Discovery Research Institute, Wako, Saitama, Japan, 3 Department of Pathology and Biology of Diseases, Graduate School of Medicine, Kyoto University, Konoe-cho, Yoshida Sakyo- ku, Kyoto, Japan Abstract Mammarian enabled (Mena), a member of the Enabled (Ena)/Vasodilator-stimulated phosphoprotein (VASP) family of proteins, has been implicated in cell motility through regulation of the actin cytoskeleton assembly, including lamellipodial protrusion. Rac1, a member of the Rho family GTPases, also plays a pivotal role in the formation of lamellipodia. Here we report that human Mena (hMena) colocalizes with Rac1 in lamellipodia, and using an unmixing assisted acceptor depletion fluorescence resonance energy transfer (u-adFRET) analysis that hMena associates with Rac1 in vivo in the glioblastoma cell line U251MG. Depletion of hMena by siRNA causes cells to be highly spread with the formation of lamellipodia. This cellular phenotype is canceled by introduction of a dominant negative form of Rac1. A Rac activity assay and FRET analysis showed that hMena knock-down cells increased the activation of Rac1 at the lamellipodia. These results suggest that hMena possesses properties which help to regulate the formation of lamellipo