GLUCOCORTICOID ACTION IN SYNCHRONIZED CELLS

同步细胞中的糖皮质激素作用

• 批准号: 3230873
• 负责人: JOHN A. CIDLOWSKI
• 金额: $ 11.24万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-08-01 至 1992-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3230873
• 关键词: HeLa cells; RNA biosynthesis; chromatin; density gradient ultracentrifugation; gel electrophoresis; gene expression; genetic manipulation; genetic regulation; genetic transcription; glucocorticoids; high performance liquid chromatography; histochemistry /cytochemistry; hormone receptor; human tissue; messenger RNA; monoclonal antibody; nucleic acid sequence; phase contrast microscopy; synchronous cell division; tissue /cell culture; tritium

The goal of our research is to continue to explore the mechanisms of glucocorticoid hormone action during the cell cycle. Work in this and other laboratories has established that the progression of cells through the cell cycle results in significant alterations in glucocorticoid receptor number and structure. We will now focus our studies on the regulation of the glucocorticoid receptor gene during the cell cycle. The availability of cDNA clones of the human glucocorticoid receptor permits us to extend our analysis of receptor regulation to the molecular level. We hypothesize that changes in cellular glucocorticoid receptor number during the cell cycle can be accounted for by alteration in receptor gene regulation. To address this question we will evaluate glucocorticoid receptor mRNA accumulation, transcription and stability during the cell cycle. Concurrently the organizational structure of the glucocorticoid receptor gene will be assessed to determine what relationship, if any, exists between receptor gene transcription and receptor gene structure. Further studies will also be initiated to identify and characterize the DNA sequences necessary for glucocorticoid receptor gene expression. Finally we seek to evaluate further a recent observation made in our laboratory which demonstrates the interaction of the human glucocorticoid receptors with the coding domain of its gene. Specifically we will characterize by several criteria the interactions between glucocorticoid receptor and its own gene and evaluate in vivo the physiological significance of such interactions. Together the studies proposed in the application should lead to a better understanding of the mechanisms involved in the regulation of human glucocorticoid receptor gene expression.

我们研究的目的是继续探索机制 糖皮质激素在细胞周期中的作用。 工作 这个和其他实验室已经确定 细胞通过细胞周期导致重大改变 糖皮质激素受体的数量和结构。 我们现在将集中精力 我们对糖皮质激素受体基因调节的研究 在细胞周期中。 cDNA克隆的可用性 人糖皮质激素受体允许我们扩展分析 受体调节至分子水平。 我们假设 在该过程中细胞糖皮质激素受体数的变化 细胞周期可以通过受体基因的改变来解释 规定。 为了解决这个问题，我们将评估 糖皮质激素受体mRNA积累，转录和 在细胞周期中的稳定性。 同时组织 糖皮质激素受体基因的结构将被评估为 确定受体基因之间存在什么关系（如果有） 转录和受体基因结构。 进一步的研究将会 也可以启动以识别和表征DNA序列 糖皮质激素受体基因表达所必需的。 最后我们 寻求进一步评估我们的最新观察 证明人相互作用的实验室 糖皮质激素受体具有其基因的编码结构域。 具体而言，我们将以几个标准来表征 糖皮质激素受体与其自身基因之间的相互作用与 在体内评估这种生理意义 互动。 一起提出的研究 应该可以更好地理解所涉及的机制 在人类糖皮质激素受体基因的调节中 表达。