Phosphoproteome and Ang II-induced VSMC Gene Expression

磷酸化蛋白质组和 Ang II 诱导的 VSMC 基因表达

• 批准号: 7025391
• 负责人: Sadashiva S Karnik
• 金额: $ 35.63万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7025391
• 关键词: actins; angiotensin II; biochemical evolution; bioinformatics; biological signal transduction; cell line; cell migration; cell proliferation; chromatin; gene expression; genetic promoter element; genetic regulatory element; mass spectrometry; muscle cells; phosphorylation; protein protein interaction; protein purification; site directed mutagenesis; small interfering RNA; transforming growth factors; vascular smooth muscle

DESCRIPTION (provided by applicant): Vascular smooth muscle cell (VSMC) phenotypes display remarkable heterogeneity in health and disease states. Yet the molecular mechanisms underlying modulation of VSMC phenotypes are not known. In Ang ll-stimulated VSMC model system, we find global changes in both the phosphoproteome and transcriptome. The cytoskeletal protein |3-actin is phosphorylated, leading to the hypothesis that phosphorylation could alter actin-dynamics, a process required for cell motility, growth and gene expression changes. Gene expression profiling indicated that several cell surface receptors, transcription regulators and proteins involved in organ damage are differentially expressed. Bioinformatics analysis of the promoter regions of these genes showed phylogenetically conserved DMA c/s-elements. We speculate that these genome regulatory elements bind transcriptional regulators, which are targets of Ang ll-induced signal transduction and alter gene expression. The short-term goals of this study are: (i) Identify phosphorylated protein targets in Ang ll-treated VSMCs by using immuno-purification and mass spectrometry (MS). We will test the network of phosphorylated proteins that modulate VSMC phenotype through gene expression changes, (ii) Determine site(s) of Ang ll- induced phosphorylation and the mechanism by which phosphorylation of p-actin affects VSMC proliferation, migration and gene expression, (iii) Define the molecular basis for Ang ll-responsiveness of genes. Our long-term goal is to understand regulation of VSMC function by angiotensin II (Ang II) and the molecular mechanism of phenotypic modulation of VSMC. Pathogenesis of major human diseases such as atherosclerosis and post-angioplasty restenosis involves VSMC phenotype switching.

描述（由申请人提供）：血管平滑肌细胞（VSMC）表型在健康和疾病状态下表现出显着的异质性。然而 VSMC 表型调节的分子机制尚不清楚。在Ang II刺激的VSMC模型系统中，我们发现磷酸化蛋白质组和转录组均发生了全局变化。细胞骨架蛋白|3-肌动蛋白被磷酸化，从而产生了这样的假设：磷酸化可以改变肌动蛋白动力学，这是细胞运动、生长和基因表达变化所需的过程。基因表达谱表明，几种细胞表面受体、转录调节因子和参与器官损伤的蛋白质存在差异表达。这些基因启动子区域的生物信息学分析显示了系统发育上保守的 DMA c/s 元件。我们推测这些基因组调控元件与转录调控因子结合，转录调控因子是 Ang II 诱导的信号转导和改变基因表达的靶标。本研究的短期目标是： (i) 通过使用免疫纯化和质谱 (MS) 鉴定 Ang II 处理的 VSMC 中的磷酸化蛋白靶标。我们将测试通过基因表达变化调节 VSMC 表型的磷酸化蛋白网络，（ii）确定 Ang II 诱导的磷酸化位点以及 p-肌动蛋白磷酸化影响 VSMC 增殖、迁移和基因表达的机制， (iii)定义基因Ang II反应性的分子基础。我们的长期目标是了解血管紧张素II（Ang II）对VSMC功能的调节以及VSMC表型调节的分子机制。动脉粥样硬化和血管成形术后再狭窄等人类主要疾病的发病机制涉及 VSMC 表型转换。