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KMID : 1140120090140020124
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Cancer Prevention Research
2009 Volume.14 No. 2 p.124 ~ p.131
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Role of Phospho-ezrin Protein in Uropod Formation of EL4 T Lymphocyte
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Kim Ji-Hyun
Kim Jeong-Hwan
Kim Gwang-Hyeon
Lee Jong-Hawn
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Abstract
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Migrating T cell shows typical hand-mirror shape, leading edge and uropod polarity. EL 4 T cells features partitioned morphology like leading edge and uropod. In order to verify this polarity, EL 4 T cells were stained for nucleus, Golgi apparatus, cell membrane with DAPI, BODIPY-FL C5-ceramide and CD44, respectively. EL4 T cells showed typical migrating T cell shape. Ezrin is a cross-linker between cell membrane and cytoskeleton. Especially, phospho-ezrin may play an important role for uropod region. Phospho-ezrin was accumulated in uropod region immune-stained with anti phospho ezrin antibody. T567D ezrin, phospho-mimetic form of ezrin, was transfected into EL4 T cells. T567D transfectants showed enhanced uropod structure and disrupted leading edge. NH2 terminal domain (NT)-ezrin transfectants showed weak uropod structure. In addition, T567D had ability of cytoplasmic accumulation of F-actin in P3X63,653 myeloma cell. To test cell biological function with enhanced uropod, cell migration assay was carried out with T567D ezrin transfectants. Reduced cell migration was detected in T567D transfectants with F2 endothelial cell, however cell migration was increased in T567D transfectants without F2 cells. Collectively, phospho-ezrin is involved in uropod fromation and uropod facilitates the rapid cell migration.
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KEYWORD
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Uropod, Phospho-ezrin, T567D-ezrin, Cell migration
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