N-terminus-mediated dim2erization of ROCK-1 is required for RhoE binding and actin reorganization

被引:20
作者
Garg, Ritu [1 ,2 ,3 ]
Riento, Kirsi [1 ,4 ]
Keep, Nicholas [2 ]
Morris, Jonathan D. H. [5 ]
Ridley, Anne J. [1 ,3 ,4 ]
机构
[1] Royal Free & Univ Coll Med Sch, Sch Med, Ludwig Inst Canc Res, London W1W 7BS, England
[2] Univ London Birkbeck Coll, Sch Crystallog, London WC1E 7HX, England
[3] Kings Coll London, Randall Div Cell & Mol Biophys, London SE1 1UL, England
[4] UCL, Dept Biochem & Mol Biol, London WC1E 6BT, England
[5] Kings Coll London, Div Canc, London SE1 1UL, England
基金
英国生物技术与生命科学研究理事会;
关键词
actin cytoskeleton; GTP-binding protein; Rho GTPase; Rho-associated kinase (ROCK); RhoE (Rnd3);
D O I
10.1042/BJ20071342
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
ROCK-I (Rho-associated kinase 1) is a serine/threonine kinase that can be activated by RhoA and inhibited by RhoE. ROCK-I has an N-terminal kinase domain, a central coiled-coil region and a RhoA-binding domain near the C-terminus. We have previously shown that RhoE binds to the N-terminal 420 amino acids of ROCK-I, which includes the kinase domain as well as N-terminal and C-terminal extensions. In the present study, we show that N-terminus-mediated dimerization of ROCK-I is required for RhoE binding. The central coiled-coil domain can also dimerize ROCK-I in cells, but this is insufficient in the absence of the N-terminus to allow RhoE binding. The kinase activity of ROCK-I(1-420) is required for dimerization and RhoE binding; however, inclusion of part of the coiled-coil domain compensates for lack of kinase activity, allowing RhoE to bind. N-terminus-mediated dimerization is also required for ROCK-I to induce the formation of stellate actin stress fibres in cells. These results indicate that dimerization via the N-terminus is critical for ROCK-I function in cells and for its regulation by RhoE.
引用
收藏
页码:407 / 414
页数:8
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