EBP50 Is Involved in the Regulation of Vascular Smooth Muscle Cell Migration and Cytokinesis
EBP50 Is Involved in the Regulation of Vascular Smooth Muscle Cell Migration and Cytokinesis
复制标题
DOI：
10.1002/jcb.23183
复制
发表时间：
2011-09-01
影响因子：
4
通讯作者：
Morel, Nicole
中科院分区：
文献类型：
--
作者：
Baeyens, Nicolas;de Meester, Carole;Morel, Nicole
Ezrin, Radixin， Moesin结合磷酸化蛋白50 （EBP50）是一种具有两个PDZ相互作用结构域的支架蛋白。我们已经证明，在去甲肾上腺素刺激的离体动脉中，EBP50与细胞骨架的几个元素相互作用。然而，EBP50对细胞骨架组织的贡献尚不清楚。我们利用原代培养的血管平滑肌细胞，研究EBP50参与细胞结构、运动和细胞周期的调控，并确定其靶蛋白及其后续作用机制。结果表明，通过siRNA转染减少EBP50可引起细胞结构的改变和细胞迁移的增加。抑制肌球蛋白IIa也能诱导相同的表型，但这种效应在EBP50缺失的细胞中不具有加性。此外，在EBP50缺失后，观察到双核细胞的比例更大，表明细胞分裂缺陷。经共免疫沉淀鉴定，EBP50可与微管蛋白和肌球蛋白IIa直接相互作用，提示EBP50可通过连接肌球蛋白IIa纤维和微管网络调节细胞迁移和细胞分裂。事实上，EBP50的缺失也会破坏肌球蛋白IIa纤维，并在片层扩张和Rac1激活中诱导稳定微管的形成。这一信号级联导致板足、尾尾的形成和局灶黏附形成的减少，引发细胞迁移。j .细胞。中国生物医学工程学报，2011,31(2):444 - 444。(C) 2011 Wiley-Liss, Inc。
Ezrin, Radixin, Moesin binding phosphoprotein 50 (EBP50) is a scaffold protein that possesses two PDZ interacting domains. We have shown that, in isolated artery stimulated with noradrenaline, EBP50 interacts with several elements of the cytoskeleton. However, the contribution of EBP50 to the organization of the cytoskeleton is unknown. We have used primary cultured vascular smooth muscle cells to investigate the involvement of EBP50 in the regulation of cell architecture, motility and cell cycle, and to identify its target proteins and subsequent action mechanism. The results showed that depletion of EBP50 by siRNA transfection induced changes in cell architecture and increased cell migration. The same phenotype was induced by inhibition of myosin IIa and this effect was not additive in cells depleted for EBP50. Moreover, a larger proportion of binucleated cells was observed after EBP50 depletion, indicating a defect in cytokinesis. The identification, after co-immunoprecipitation, of a direct interaction of EBP50 with both tubulin and myosin IIa suggested that EBP50 could regulate cell migration and cytokinesis by linking myosin IIa fibers and microtubule network. Indeed, depletion of EBP50 also dismantled myosin IIa fibers and induced the formation of stable microtubules in lamellae expansions and Rac1 activation. This signaling cascade leads to the formation of lamellipodia, trailing tails and decrease of focal adhesion formation, triggering cell migration. J. Cell. Biochem. 112: 2574-2584, 2011. (C) 2011 Wiley-Liss, Inc.