DGCR6L, a novel PAK4 interaction protein, regulates PAK4-mediated migration of human gastric cancer cell via LIMK1
DGCR6L, a novel PAK4 interaction protein, regulates PAK4-mediated migration of human gastric cancer cell via LIMK1
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DGCR6L 是一种新型 PAK4 相互作用蛋白，通过 LIMK1 调节 PAK4 介导的人胃癌细胞迁移
DOI：
10.1016/j.biocel.2009.09.008
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发表时间：
2010-01-01
影响因子：
4
通讯作者：
Li, Feng
中科院分区：
文献类型：
--
作者：
Li, Xiaodong;Ke, Qiang;Li, Feng
p21激活激酶4（PAK 4）在多种人类肿瘤中存在过表达、基因扩增和突变。PAK 4通过磷酸化LIMK 1调节肌动蛋白骨架重组，促进癌细胞迁移。利用酵母双杂交筛选，我们鉴定了一种新的PAK 4结合蛋白DGCR 6L，它与癌细胞转移有关。我们通过GST pull-down实验证实PAK 4与DGCR 6L特异性结合，并通过免疫沉淀在哺乳动物细胞中发现内源性PAK 4与DGCR 6L之间的关联。此外，DGCR 6L的L115是结合PAK 4的C-末端的466- 572 aa的关键氨基酸。重要的是，DGCR 6L是PAK 4-DGCR 6L-beta-actin复合物形成所必需的。DGCR 6L过表达可促进PAK 4介导的AGS细胞迁移，而DGCR 6L敲低则可显著抑制AGS细胞迁移。此外，DGCR 6L（L115 V）不与PAK 4结合，失去了促进AGS细胞迁移的能力。DGCR 6L与PAK 4或F-actin共定位，并以剂量依赖性方式增强LIMK 1和cofilin的磷酸化水平。综上所述，我们的结果表明，DGCR 6L，一种新的PAK 4相互作用蛋白，通过LIMK 1调节PAK 4介导的人胃癌细胞迁移。(C)2009爱思唯尔有限公司保留所有权利。
Overexpression, genetic amplification and mutations of p21-activated kinase 4 (PAK4) were found in a variety of human cancers. PAK4 regulated actin cytoskeleton reorganization by phosphorylating LIMK1 and promoted cancer cells migration. Using yeast two-hybrid screen, we identified a novel PAK4 binding protein, DGCR6L, which was associated with cancer cell metastasis. We confirmed PAK4 binding to the DGCR6L specifically by GST pull-down assay, and found an association between endogenous PAK4 and DGCR6L by immunoprecipitation in mammalian cells. Furthermore, L115 of DGCR6L was the critical amino acid to bind 466-572aa in the very C-terminus of PAK4. Importantly, DGCR6L was required for the formation of PAK4-DGCR6L-beta-actin complex. Overexpressed DGCR6L promoted migration of AGS cells mediated by PAK4, whereas knock-down of DGCR6L markedly inhibited the migration of those cells. Moreover, DGCR6L (L115V), which did not bind to PAK4, lost the ability to promote AGS cells migration. DGCR6L colocalized with PAK4 or F-actin and enhanced the phosphorylation level of LIMK1 and cofilin in a dose dependent manner. Taken together, our results demonstrated that DGCR6L, a novel PAK4 interacting protein, regulated PAK4-mediated migration of human gastric cancer cells via LIMK1. (C) 2009 Elsevier Ltd. All rights reserved.