ERK1/2 Associates with the c-Met-binding Domain of Growth Factor Receptor-bound Protein 2 (Grb2)-associated Binder-1 (Gab1)
ERK1/2 Associates with the c-Met-binding Domain of Growth Factor Receptor-bound Protein 2 (Grb2)-associated Binder-1 (Gab1)
复制标题
ERK1/2 与生长因子受体结合蛋白 2 (Grb2) 相关 Binder-1 (Gab1) 的 c-Met 结合域相关
DOI：
--
复制
发表时间：
2004
影响因子：
4.8
通讯作者：
J. Abe
中科院分区：
文献类型：
--
作者：
M. Osawa;Seigo Itoh;S. Ohta;Qunhua Huang;B. Berk;Nicole;W. Che;B. Ding;C. Yan;J. Abe
血管成形术或动脉粥样硬化斑块破裂后，内皮细胞（EC）迁移有助于再内皮化。细胞外信号调节激酶（ERK）1/2在再内皮化过程中易位至细胞核并激活转录因子如Ets样转录因子-1和早期生长反应因子-1（Egr-1）。由于ERK 1/2不具有核定位信号（NLS），其在细胞核中的移位和积聚机制尚不清楚。由于Gab 1在其N-末端区域具有假定的NLS，并且Gab 1与磷酸化的ERK 1/2相关联，因此我们假设Gab 1参与ERK 1/2和Egr-1的核积累。使用再生EC作为模型系统，我们发现，内源性生长因子受体结合蛋白2相关的结合剂1（Gab 1）易位到细胞核中迁移EC。野生型红色荧光蛋白标记的Gab 1可在细胞核和细胞质中观察到，而推定的NLS缺失突变体（Δ NLS-Gab 1）特异性地定位于细胞质中。此外，通过反义Gab 1寡核苷酸或过表达Δ NLS-Gab 1降低Gab 1表达可抑制血清诱导的ERK 1/2和Egr-1核积聚，表明Gab 1的NLS功能作用以及Gab 1-ERK 1/2相互作用在ERK 1/2-Egr-1核积聚中的作用。为了研究Gab 1-ERK 1/2相互作用是否对ERK 1/2和Egr-1核积累至关重要，我们创建了由Gab 1的c-Met结合结构域（氨基酸442-536）组成的显性负性Gab 1构建体。我们发现，过度表达c-Met结合域的Gab 1破坏血清诱导的Gab 1-ERK 1的相互作用，抑制ERK 1和Egr-1的核积累。这些数据表明，Gab 1-ERK 1/2结合和核转位在Egr-1核积累中起着至关重要的作用。
Endothelial cell (EC) migration contributes to reendothelialization after angioplasty or rupture of atherosclerotic plaques. Extracellular signal-regulated kinase (ERK)1/2 translocates to the nucleus and activates transcription factors such as Ets-like transcription factor-1 and early growth response factor-1 (Egr-1) during reendothelialization. Because ERK1/2 does not possess a nuclear localization signal (NLS), its mechanism of translocation and accumulation in the nucleus remains unclear. Because Gab1 has a putative NLS in its N-terminal region, and Gab1 associates with phosphorylated ERK1/2, we hypothesized that Gab1 participates in ERK1/2 and Egr-1 nuclear accumulation. Using regenerating EC as a model system, we found that endogenous growth factor receptor-bound protein 2-associated binder-1 (Gab1) translocates into the nucleus in migrating EC. Wild-type red fluorescent protein-tagged Gab1 could be observed in both nucleus and cytoplasm, whereas the putative NLS deletion mutant (ΔNLS-Gab1) specifically localized in the cytoplasm. In addition, reduction of Gab1 expression by antisense Gab1 oligos or overexpression of ΔNLS-Gab1 inhibited serum-induced ERK1/2 and Egr-1 nuclear accumulation, suggesting a functional role for the NLS of Gab1 and a role for Gab1-ERK1/2 interactions in ERK1/2-Egr-1 nuclear accumulation. To investigate whether Gab1-ERK1/2 interaction is critical for ERK1/2 and Egr-1 nuclear accumulation, we created a dominant-negative Gab1 construct that consisted of the c-Met binding domain (amino acids 442-536) of Gab1. We found that overexpression of the c-Met binding domain of Gab1 disrupted serum-induced Gab1-ERK1 interaction and inhibited ERK1 and Egr-1 nuclear accumulation. These data suggest that Gab1-ERK1/2 binding and their nuclear translocation play a crucial role in Egr-1 nuclear accumulation.
DOI：
10.1161/01.atv.17.10.2280
发表时间：
1997-10-01
影响因子：
8.7
作者：
Khachigian, LM;Anderson, KR;Collins, T
通讯作者：
Collins, T