Endoglin regulates cytoskeletal organization through binding to ZRP-1, a member of the Lim family of proteins
Endoglin regulates cytoskeletal organization through binding to ZRP-1, a member of the Lim family of proteins
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DOI：
10.1074/jbc.m400843200
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发表时间：
2004-07-30
影响因子：
4.8
通讯作者：
Bernabéu, C
中科院分区：
文献类型：
--
作者：
Sanz-Rodriguez, F;Guerrero-Esteo, M;Bernabéu, C
内皮糖蛋白是在内皮细胞表面大量表达的转化生长因子-β受体复合物的组分，并且在心血管发育和血管重塑中起重要作用。通过使用endoglin的胞质结构域作为诱饵筛选蛋白质相互作用，我们已经确定了ZRP-1（zyxin-related protein 1），一个476个氨基酸的成员，属于一个家庭的LIM包含蛋白质，包括zyxin和脂肪瘤首选的合作伙伴。内皮糖蛋白相互作用区域被定位在ZRP-1C末端的三个双锌指LIM结构域内。对ZRP-1的亚细胞分布的分析表明，在内皮糖蛋白不存在的情况下，ZRP-1主要定位于粘着斑位点，而在内皮糖蛋白存在的情况下，发现ZRP-1沿着肌动蛋白应力纤维。由于LIM蛋白家族已被证明与肌动蛋白细胞骨架相关，我们研究了内皮糖蛋白对这种结构的调节作用的可能性。内皮糖蛋白的表达导致肌动蛋白细胞骨架的戏剧性重组。在内皮糖蛋白的情况下，F-肌动蛋白被定位到密集的聚集束，而在内皮糖蛋白的存在下，在内皮细胞中表达，F-肌动蛋白是在应力纤维和共定位与ZRP-1。此外，小干扰RNA介导的抑制内皮糖蛋白或ZRP-1，或内皮糖蛋白在内皮细胞中的聚集，导致F-肌动蛋白纤维的错误定位。这些结果表明，内皮糖蛋白的调节作用，通过其与ZRP-1的相互作用，在肌动蛋白细胞骨架组织。
Endoglin is a component of the transforming growth factor-beta receptor complex abundantly expressed at the surface of endothelial cells and plays an important role in cardiovascular development and vascular remodeling. By using the cytoplasmic domain of endoglin as a bait for screening protein interactors, we have identified ZRP-1 (zyxin-related protein 1), a 476-amino acid member that belongs to a family of LIM containing proteins that includes zyxin and lipoma-preferred partner. The endoglin interacting region was mapped within the three double zinc finger LIM domains of the ZRP-1 C terminus. Analysis of the subcellular distribution of ZRP-1 demonstrated that in the absence of endoglin, ZRP-1 mainly localizes to focal adhesion sites, whereas in the presence of endoglin ZRP-1 is found along actin stress fibers. Because the LIM family of proteins has been shown to associate with the actin cytoskeleton, we investigated the possibility of a regulatory role for endoglin with regard to this structure. Expression of endoglin resulted in a dramatic reorganization of the actin cytoskeleton. In the absence of endoglin, F-actin was localized to dense aggregates of bundles, whereas in the presence of endoglin, expressed in endothelial cells, F-actin was in stress fibers and colocalized with ZRP-1. Furthermore, small interfering RNA-mediated suppression of endoglin or ZRP-1, or clustering of endoglin in endothelial cells, led to mislocalization of F-actin fibers. These results suggest a regulatory role for endoglin, via its interaction with ZRP-1, in the actin cytoskeletal organization.