CREB mediates UTP-directed arterial smooth muscle cell migration and expression of the chemotactic protein osteopontin via its interaction with activator protein-1 sites
CREB mediates UTP-directed arterial smooth muscle cell migration and expression of the chemotactic protein osteopontin via its interaction with activator protein-1 sites
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DOI：
10.1161/01.res.0000266609.28312.de
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发表时间：
2007-05-11
影响因子：
20.1
通讯作者：
Desgranges, Claude
中科院分区：
文献类型：
--
作者：
Jalvy, Sandra;Renault, Marie-Ange;Desgranges, Claude
转录因子 cAMP 反应元件结合蛋白 (CREB) 被发现参与动脉平滑肌细胞 (SMC) 迁移。我们之前证明骨桥蛋白（OPN）表达是UTP介导的动脉SMC迁移的关键步骤，并且激活蛋白（AP）-1、核因子κB和上游刺激转录因子参与该OPN表达。本研究旨在确定 CREB 在 UTP 诱导的培养 SMC 迁移和 OPN 表达中的作用。我们发现，CREB 是由 UTP 通过细胞外信号调节激酶 1/2 和 p38 丝裂原激活蛋白激酶途径激活的，而不是由蛋白激酶 A 激活的。显性失活 CREB 的过表达和 CREB 小干扰 RNA 处理均抑制 UTP 诱导的 OPN 表达和 SMC 迁移。凝胶迁移和染色质免疫沉淀测定表明，CREB 结合 OPN 启动子上的 2 个 AP-1 位点（-1870 和 -76）和一个 cAMP 响应元件样位点（-1403）。这些位点的突变表明，UTP 诱导的 OPN 表达仅需要 2 个 AP-1 位点。此外，凝胶超移和连续染色质免疫沉淀分析表明 CREB 与 OPN 启动子 AP-1 位点上的 c-Fos 相关。这些结果表明，CREB 通过与 2 个 AP-1 位点结合参与诱导 UTP 激活的 OPN 表达，从而参与 UTP 介导的 SMC 迁移。
The transcription factor cAMP responsive element-binding protein (CREB) has been found to be involved in arterial smooth muscle cell (SMC) migration. We previously demonstrated that osteopontin (OPN) expression is a key step for UTP-mediated migration of arterial SMCs and that activator protein (AP)-1, nuclear factor kappa B, and upstream stimulatory transcription factors are involved in this OPN expression. The present study aims to determine the role of CREB in UTP-induced migration and OPN expression in cultured SMCs. We found that CREB is activated by UTP via extracellular signal- regulated kinase 1/2 and p38 mitogen-activated protein kinase pathways but not by protein kinase A. Both overexpression of a dominant negative CREB and CREB small interfering RNA treatment suppressed UTP-induced OPN expression and SMC migration. Gel-shift and chromatin immunoprecipitation assays revealed that CREB binds 2 AP-1 sites (-1870 and -76) and a cAMP responsive element-like site (-1403) on the OPN promoter. Mutations of these sites showed that only the 2 AP-1 sites were required for UTP-induced OPN expression. Moreover, gel-supershift and sequential chromatin immunoprecipitation assays suggested that CREB was associated with c-Fos on the AP-1 sites of the OPN promoter. These results demonstrate that CREB participates in the induction of UTP-activated OPN expression via its binding to 2 AP-1 sites and is thus involved in UTP-mediated SMC migration.