Reg of SM22 Alpha Transcription in Smooth Muscle Cells

平滑肌细胞中 SM22 Alpha 转录的调节

• 批准号: 6332279
• 负责人: Michael S Parmacek
• 金额: $ 27.74万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6332279
• 关键词: biological signal transduction; cell differentiation; developmental genetics; gene expression; genetic promoter element; genetic regulation; genetic regulatory element; genetic transcription; genetically modified animals; homeobox genes; laboratory mouse; laboratory rat; protein protein interaction; transcription factor; vascular smooth muscle

DESCRIPTION (Applicant's abstract): The capacity of vascular smooth muscle cells (SMCs) to modulate their response to arterial injury has been implicated in the pathogenesis of vascular proliferative syndromes including atherosclerosis and restenosis following coronary intervention. We have utilized the SMC-specific mouse SM22a promoter as a model system to elucidate the transcriptional programs that control VSMC development and differentiation. During the initial funding cycle of this award, the cis-acting elements and trans-acting factors that control expression of the SM22a promoter were defined. Two nuclear protein binding sites in the SM22a promoter (SME- 1 and SME-4) were identified which contain consensus CArU boxes that bind specifically to the MADS box transcription factor, SRF. Remarkably, a multimerized copy of either SME-l or SME-4 is necessary and sufficient to restrict activity of a LacZ reporter gene to arterial SMCs in transgenic mice. These data lead us to hypothesize that SRF, in concert with other potentially novel transcriptional activators (and potentially repressors), regulates SMC-specific gene expression and SMC phenotype. The overall goal of the proposed studies is to elucidate the molecular basis of SRF-dependent transcription in arterial SMCs. The specific aims of these studies are to: i) Examine the molecular basis of SRF-CArG box activity in SMCs, ii) Examine protein-protein associations that modulate activity of the SM22a promoter in SMCs with particular attention on SRF/homeobox protein interactions, iii) Examine signaling pathways underlying LIMK- 1-mediated activation of the SM22cz promoter, and iv) Examine the capacity of SRF-/- ES cells to contribute to the SMC lineage(s) in SRF-/- -C57BL/6 chimenc mice and define downstream SMC and mesodermal genes regulated by SRF in the embryo. Taken together, these studies will elucidate the transcriptional program and an important signaling pathway that modulates expression of the SM22a gene in SMCs. Because CArG box-containing elements have been identified in multiple other SMC-specific genes these studies will provide fundamental insights into the molecular mechanisms that control SMC differentiation and the modulation of vascular SMC phenotype.

描述（申请人的摘要）：血管平滑肌的能力 已经暗示了细胞（SMC）调节其对动脉损伤的反应 在血管增生综合症的发病机理中 冠状动脉干预后动脉粥样硬化和再狭窄。我们有 利用SMC特异性小鼠SM22A启动子作为模型系统阐明 控制VSMC开发和差异化的转录程序。 在该奖项的最初资助周期中，顺式作用要素和 控制SM22A启动子表达表达的跨作用因子是 定义。 SM22A启动子中的两个核蛋白结合位点（SME-1和 识别了SME-4），其中包含绑定的共识盒 特别针对MADS盒转录因子SRF。值得注意的是， SME-L或SME-4的多层次副本是必要的，足以 将LACZ报告基因的活性限制为转基因小鼠中的动脉SMC。 这些数据使我们假设SRF与其他潜在的可能 新颖的转录激活剂（和潜在的阻遏物）调节 SMC特异性基因表达和SMC表型。总体目标 拟议的研究是阐明依赖SRF的分子基础 动脉SMC中的转录。这些研究的具体目的是：i） 检查SMC中SRF-CARG盒活性的分子基础，ii）检查 调节SM22A启动子活性的蛋白质 - 蛋白质关联 特别注意SRF/同源蛋白质相互作用的SMC，iii） 检查SM22CZ的Limk-1介导的激活的信号通路 启动子和iv）检查SRF - / - ES细胞有助于 SRF - / - -C57BL/6 Chimenc小鼠中的SMC谱系（S），并定义下游SMC和 胚胎中SRF调控的中胚层基因。总之，这些研究 将阐明转录程序和重要的信号通路 这调节了SMCS中SM22A基因的表达。因为卡格 在其他多个SMC特定的特定于 基因这些研究将为分子提供基本见解 控制SMC分化和血管SMC调节的机制 表型。