Transcriptional regulation of CRP1 in smooth muscle

平滑肌中 CRP1 的转录调控

• 批准号: 7171602
• 负责人: Brenda J Lilly
• 金额: $ 28.51万
• 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/7171602
• 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; Personal Satisfaction; Phenotype; Range; 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

DESCRIPTION (provided by applicant): 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-¿), use a distinct combination of intracellular pathways and transcription factors to impart smooth muscle-restricted expression. TGF-¿ 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 (TGF-¿1). 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.

描述（申请人提供）：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。这些细胞表现出一系列依赖于转录程序选择性利用的表型。虽然平滑肌调节对正常血管功能至关重要，但它们的适应能力与血管闭塞疾病的病理相反。因此，控制基因转录导致结果表型的分子机制是生物学和临床相关的。为了阐明控制平滑肌转录的关键特征，我们的研究重点是前体细胞向成熟平滑肌的发育和分化。分化伴随着收缩所需的一组精确基因的精心激活。虽然明确的调控元件和相应的转录因子被认为在分化中发挥作用，但它们如何与传递选择性基因表达的信号耦合尚不清楚。我们假设细胞外信号，如转化生长因子- β （TGF-¿），使用细胞内途径和转录因子的独特组合来传递平滑肌受限表达。TGF-¿已成为控制平滑肌细胞表型的主要候选者。本研究的目的是利用富半胱氨酸蛋白1 （CRP1）基因的平滑肌特异性表达来研究在特定信号事件中传递选择性转录活性的顺式作用元件和反式作用因子。我们已经确定了CRP1基因的一个独特的调控区域，该区域仅在动脉平滑肌细胞中驱动表达。我们打算使用这个调控元件来表征分化平滑肌细胞中CRP1基因表达的重要转录途径。具体目的是：1)明确转化生长因子- β -1 （TGF-¿1）激活CRP1基因的转录机制。2)确定血清反应因子（SRF）的功能活性与控制平滑肌基因表达的信号事件之间的关系。3)明确通过靶向诱变确定CArG元件在CRP1表达中的必要性。这些研究具有重要意义，因为它们将超越转录调节因子的分析，并确定介导转激活的确切途径，以解决有关平滑肌细胞表型表现的基本问题。