The molecular mechanism of p160ROCK-mediated signaling pathway

p160ROCK介导的信号通路的分子机制

基本信息

批准号：
10670120
负责人：
ISHIZAKI Toshimasa
金额：
$ 1.79万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

项目摘要

We found Rho-associated coiled coil forming protein kinase (p160ROCK) is serine/threonine kinase. p160ROCK is a member of effector protein of small GTPase Rho and specisically binds to activated form of Rho thereby becomes activated upon this binding. During transfction experiments in fibroblast and eptherial cells, this kinase is involved in formation of actin stress fibers and focal adhesions mediated by Rho. In addition to this, in nuroblastama N1E-115 cell this kinase induces cell rounding by regulating contractile force at cell cortex.Thus, it is thought that p160ROCK is localized to several specific sites in the cell and phosphorylates several substrates.Recently, we developed a specific inhibitor of ROCK, we named it Y-27632. This compound competes with ATP at the kinase domain. Using this compound, we reported that Rho/ROCK pathway regulates blood pressure in hypertension model rats, indicating that this pathway is involved in the pathophysiology of hypertension. However, the molecular mechanism of Rho/ROCK pathway is not clearly known.In this study, we try to understand the molecular mechanism (s) of p160ROCK-mediated signaling pathway (s) through the isolation of phosphorylated substrates for p160ROCK.As a result, our major findings are as follows,1. p160ROCK regulates Confilin/ADF phosphorylaytion through phosphorylation and activation of LIM-kinase.2. p160ROCK cooperates with mDia, another effector molecule of Rho, on Rho-mediated actin stress fiber formation and focal adhesion formation.

我们发现Rho相关卷曲螺旋形成蛋白激酶（p160ROCK）是丝氨酸/苏氨酸激酶。 p160ROCK 是小 GTP 酶 Rho 效应蛋白的成员，与 Rho 的活化形式特异性结合，从而在这种结合后被激活。在成纤维细胞和上皮细胞的转染实验中，该激酶参与 Rho 介导的肌动蛋白应力纤维和粘着斑的形成。除此之外，在神经母细胞瘤 N1E-115 细胞中，该激酶通过调节细胞皮质的收缩力来诱导细胞变圆。因此，人们认为 p160ROCK 定位于细胞中的几个特定位点并磷酸化多个底物。最近，我们开发了一种ROCK的特异性抑制剂，我们将其命名为Y-27632。该化合物在激酶结构域与 ATP 竞争。使用该化合物，我们报道了 Rho/ROCK 通路调节高血压模型大鼠的血压，表明该通路参与高血压的病理生理学。然而，Rho/ROCK通路的分子机制尚不清楚。在本研究中，我们试图通过分离p160ROCK的磷酸化底物来了解p160ROCK介导的信号通路的分子机制。，我们的主要发现如下：1． p160ROCK通过LIM激酶的磷酸化和激活来调节Confilin/ADF磷酸化。2． p160ROCK 与 Rho 的另一种效应分子 mDia 合作，共同参与 Rho 介导的肌动蛋白应力纤维形成和粘着斑形成。