Identification of a cell polarity-related protein, Lin-7B, as a binding partner for a Rho effector, Rhotekin, and their possible interaction in neurons
Identification of a cell polarity-related protein, Lin-7B, as a binding partner for a Rho effector, Rhotekin, and their possible interaction in neurons
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DOI：
10.1016/j.neures.2006.08.003
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发表时间：
2006-12-01
影响因子：
2.9
通讯作者：
Nagata, Koh-ichi
中科院分区：
文献类型：
--
作者：
Sudo, Kaori;Ito, Hidenori;Nagata, Koh-ichi
Rhotekin是Rho的效应器，在大脑中高度表达，但其在神经元中的功能(S)几乎未知。为了确定Rhotekin在神经细胞中的性质，我们重点研究了它与极性相关分子的相互作用。在本研究中，我们通过酵母双杂交筛选人脑cDNA文库，确定了PDZ蛋白LIN-7B作为Rhotekin的结合伙伴。然后，我们发现Rhotekin在体外下拉试验中与Lin-7B相互作用，并在COS7细胞和大鼠大脑中形成免疫复合体。Rhotekin的C端3个氨基酸是与LIN-7B相互作用所必需的。在COS7细胞表达系统中，当活性RhoA存在时，它们的结合亲和力增强。此外，免疫组织化学分析表明，LIN-7和Rhotekin在神经元中都有丰富的表达。这些结果表明，LIN-7通过Rho/Rhotekin信号在神经元功能中发挥一定的作用。(C)2006年爱思唯尔爱尔兰有限公司和日本神经科学学会。版权所有。
Rhotekin, an effector of Rho, is highly expressed in the brain but its function(s) in neurons is almost unknown. In an attempt to define the properties of Rhotekin in neuronal cells, we focused on its interaction with polarity-related molecules. In the present study, we identified a PDZ protein, Lin-7B, as a binding partner for Rhotekin by yeast two-hybrid screening of human brain cDNA library. We then found that Rhotekin interacts with Lin-7B in in vitro pull-down assays, and forms an immunocomplex in COS7 cells and the rat brain. The C-terminal three amino acids of Rhotekin were essential for the interaction with Lin-7B. Their binding affinity became increased in the presence of active RhoA in the COS7 cell expression system. In addition, immunohistochemical analyses demonstrated that Lin-7 as well as Rhotekin is enriched in neurons. These results suggest that Lin-7 plays some role in neuronal functions in concert with Rho/Rhotekin signals. (c) 2006 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.