ALPHA-2 ADRENERGIC RECEPTOR GENE TRANSCRIPTION

ALPHA-2 肾上腺素受体基因转录

• 批准号: 2224244
• 负责人: DIANE E HANDY
• 金额: $ 11.61万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-01-01 至 1997-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2224244
• 关键词: DNA binding protein; DNA footprinting; alpha adrenergic receptor; blood pressure; brain; cyclic AMP; gel mobility shift assay; genetic regulatory element; genetic transcription; kidney; laboratory rat; receptor expression; tissue /cell culture; transcription factor

Alpha 2 adrenergic receptors have been shown to play an important role in homeostasis of kidney function and in the regulation of the central sympathetic system. Dysfunction of these receptors is probably involved in the development and/or maintenance of some forms of hypertension. Recent molecular analysis has shown that there are three subtypes of alpha 2 adrenergic receptors, each encoded by a separate gene in rat and human. The long term goal of this project is to understand the relative contributions of each subtype to cardiovascular function and blood pressure regulation. It has been shown that each of these receptor subtypes has a different pattern of tissue specific expression. It is our hypothesis that the diversity of alpha 2 adrenergic receptor genes allows for the tissue specific expression of genes that differ in their transcriptional regulation and vary in their response to physiological stimuli. It is the goal of this grant to understand the mechanism of tissue specific expression of various alpha 2 genes and to begin to analyze the intracellular and extracellular signals that regulate alpha 2 gene expression. The following specific aims will be undertaken: (1) to study the distribution of transcripts specific for each of the three alpha 2 genes in subregions of brain and kidney; (2) To study the cis regulatory regions of the various alpha 2 genes that may be responsible for their differential expression by comparing the ability of putative regulatory regions to direct transcription in a variety of cell lines. (3) To characterize nuclear factors that are important or tissue specific expression of a alpha 2 adrenergic receptor genes by analyzing DNA/nuclear factor interactions in DNase I footprinting and gel mobility shift assays, by assessing the effects of point mutations in nuclear factor binding sites to alter transcription and by studying the tissue distribution of key nuclear factors. (4) To study intracellular and extracellular signals that alter alpha 2 gene transcription by monitoring the ability of agents, such as cAMP or receptor agonists, to alter transcription in various cells lines and to study the cis regulatory elements and trans-acting factors that mediate these affects by methods in specific aims 2 and 3. These studies will lead to the development of intraventions to specifically affect the level of expression of each alpha 2 gene at the level of transcription. In summary, the proposed studies will provide important information concerning the expression and regulation of genes that encode the receptors that have many important biological functions.

α2肾上腺素能受体已被证明发挥着重要作用 肾功能的稳态和中枢的调节 交感神经系统。 这些受体的功能障碍可能与 某些形式的高血压的发展和/或维持。 最近的分子分析表明，存在三种亚型 α2 肾上腺素能受体，每个受体在大鼠和 人类。 该项目的长期目标是了解相对 每种亚型对心血管功能和血液的贡献 压力调节。 已经表明，这些受体中的每一个 亚型具有不同的组织特异性表达模式。 这是 我们的假设是α2肾上腺素受体基因的多样性 允许不同基因的组织特异性表达 转录调控及其对生理反应的不同 刺激。 这项资助的目标是了解这一机制 各种α2基因的组织特异性表达并开始 分析调节α的细胞内和细胞外信号 2基因表达。 将实现以下具体目标：（1） 研究三者中每一个特定转录本的分布 大脑和肾脏亚区域的 α2 基因； （二）研究顺式 可能负责的各种α2基因的调节区域 通过比较假定的能力来确定它们的差异表达 调控区域在多种细胞系中指导转录。 (3) 表征重要的或组织特异性的核因子 通过分析α2肾上腺素受体基因的表达 DNase I 足迹和凝胶迁移率中 DNA/核因子的相互作用 转变分析，通过评估核中点突变的影响 因子结合位点来改变转录并通过研究组织 关键核因子的分布。 (4) 研究细胞内和 通过监测改变 α2 基因转录的细胞外信号 药物（例如 cAMP 或受体激动剂）改变的能力 各种细胞系中的转录并研究顺式调控 通过方法介导这些影响的元素和反式作用因子 具体目标 2 和 3。 这些研究将导致干预措施的发展 特别影响每个α2基因的表达水平 转录水平。 总之，拟议的研究将提供 有关基因表达和调控的重要信息 编码具有许多重要生物学功能的受体。