Transcriptional regulation of CRP1 in smooth muscle

平滑肌中 CRP1 的转录调控

• 批准号: 7582284
• 负责人: Brenda J Lilly
• 金额: $ 28.55万
• 依托单位: AUGUSTA UNIVERSITY
• 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/7582284
• 关键词: Address; Be++ element; Beryllium; Binding Sites; Blood Vessels; Boxing; Cells; Coupled; Data; Development; Disease; Elements; Embryonic Development; Enhancers; Event; Fibroblasts; Gene Activation; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Growth Factor; Indium; Link; Mediating; Molecular; Muscle function; Mutagenesis; Nucleic Acid Regulatory Sequences; Participant; Pathology; Pathway interactions; Phenotype; Regulation; Regulatory Element; Reporter Genes; Research Personnel; Role; Serum Response Factor; Signal Pathway; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Testing; Trans-Activators; Transactivation; Transcriptional Activation; Transcriptional Regulation; Transforming Growth Factor beta; cis acting element; clinically relevant; cysteine rich protein; design; extracellular; human TGFB1 protein; innovation; novel; precursor cell; programs; research study; response; tool; transcription factor

Vascular smooth muscle cells are a vital component of the blood vessel wall, possessing extraordinary adaptive abilities. These cells display a range of phenotypes that are dependent upon the selective utilization of transcriptional programs. While smooth muscle modulation is essential for normal blood vessel function, their adaptive abilities are adversely associated with the pathologies of vascular occlusion diseases. Consequently, the molecular mechanisms governing gene transcription leading to resultant phenotypes are both biologically and clinically relevant. As an inroad to elucidating critical features that control smooth muscle transcription, our studies are focused on the development and differentiation of precursor cells into mature smooth muscle. Differentiation is accompanied by the orchestrated activation of a precise set of genes required for contraction. Though definitive regulatory elements, and corresponding transcription factors are recognized to have a role in differentiation, how they are coupled to signals that convey selective gene expression is not well understood. We hypothesize that extracellular signals, like transforming growth factor-beta (TGF-IS), use a distinct combination of intracellular pathways and transcription factors to impart smooth muscle-restricted expression. TGF-li has emerged as primary candidate for governing smooth muscle cell phenotypes. The aims of this proposal are designed to employ the smooth muscle-specific expression of the cysteine-rich protein 1 (CRP1) gene to investigate the cis-acting elements, and trans- acting factors that convey selective transcriptional activity in response to defined signaling events. We have identified a unique regulatory region of the CRP1 gene that drives expression exclusively in arterial smooth muscle cells. We intend to use this regulatory element to characterize essential transcriptional pathways important for the expression of the CRP1 gene in differentiated smooth muscle cells. The specific aims are: 1) To define the transcriptional mechanisms underlying the activation of the CRP1 gene by transforming growth factor-beta-1 (TOF-I^). 2) To determine the relationship between the functional activity of serum response factor (SRF), and signaling events that govern smooth muscle gene expression. 3) To unequivocally determine the necessity of the CArG element for expression of CRP1 by targeted mutagenesis. These studies are significant, as they will extend beyond the analysis of the transcriptional regulators, and determine the exact pathways that mediate transactivation, to address fundamental questions regarding the manifestation of smooth muscle cell phenotypes.

血管平滑肌细胞是血管壁的重要组成部分，具有非凡的 适应能力。这些细胞显示出一系列取决于选择性利用的表型 转录程序。虽然平滑肌调节对于正常血管功能是必不可少的， 它们的适应能力与血管闭塞疾病的病理学相反地相关。 因此，控制基因转录导致表型的分子机制是 生物学和临床相关性。作为阐明控制平滑的关键特征的进展， 肌肉转录，我们的研究集中在前体细胞的发育和分化， 成熟平滑肌分化伴随着一组精确的 收缩所需的基因。虽然限定性调控元件和相应的转录 因素被认为在分化中起作用，它们如何与传达选择性的信号相结合， 基因表达还不清楚。我们假设细胞外信号，如转化生长 因子-β（TGF-1 S），使用细胞内途径和转录因子的独特组合来赋予 平滑的肌肉限制的表达。TGF-李已成为执政顺利的主要候选人 肌细胞表型该提案的目的是采用平滑肌特异性 富含半胱氨酸蛋白1（CRP 1）基因的表达，以研究顺式作用元件，和反式作用元件。 作用因子传递选择性转录活性以响应确定的信号事件。我们有 确定了CRP 1基因的一个独特的调控区域，该区域仅在动脉平滑肌中驱动表达， 肌肉细胞我们打算用这个调控元件来表征重要的转录途径 对于分化的平滑肌细胞中CRP 1基因的表达很重要。具体目标是： 1)为了确定CRP 1基因激活的转录机制，通过转化 生长因子-β-l（TOF-I^）。2)确定血清功能活性与 反应因子（SRF）和控制平滑肌基因表达的信号事件。3)到 明确地确定CArG元件对于通过靶向表达CRP 1的必要性， 诱变这些研究是重要的，因为它们将超越转录的分析。 调节因子，并确定介导反式激活的确切途径，以解决基本的 关于平滑肌细胞表型表现的问题。