MOLECULAR BIOLOGY OF STEROID HORMONE ACTION

类固醇激素作用的分子生物学

• 批准号: 3273331
• 负责人: GORDON M RINGOLD
• 金额: $ 17.02万
• 依托单位: SYNTEX (USA), INC.-RESEARCH DIVISION
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-12-01 至 1991-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3273331
• 关键词: DNA binding protein; Escherichia coli; binding proteins; complementary DNA; cytochrome P450; endonuclease; eukaryote; flow cytometry; gene expression; genetic manipulation; genetic recombination; genetic transcription; glucocorticoids; hepatocellular carcinoma; hormone receptor; hormone regulation /control mechanism; immunochemistry; laboratory mouse; laboratory rabbit; laboratory rat; liver; messenger RNA; microorganism genetics; molecular cloning; monoclonal antibody; mouse mammary tumor virus; nucleic acid hybridization; nucleic acid sequence; radiotracer; spectrometry; steroid hormone; temperature sensitive mutant; vaccinia virus; viral carcinogenesis; virus RNA

Virtually all mammalian cells contain receptors for one or more classes of steroid hormones. These high-affinity receptor proteins most likely mediate the major effects of steroids on cell function by interacting with specific DNA sequences in (or near) target genes. Upon so doing, the steroid-receptor complex stimulates initiation of transcription thereby leading to increased production of specific RNAs. The study of complex regulatory mechanisms in higher eukaryotes has been hampered by the difficulty in manipulating such organisms genetically and by the enormity of mammalian genomes. We therefore have turned to simplified tissue culture systems and isolated genes to study the molecular mechanisms by which steroid hormones regulate gene expression. We will use genetic biochemical and molecular approaches to investigate the following issues: 1) What is the structure of the glucocorticoid receptor and which portions of the protein are required for its hormone binding, DNA binding, and transcription stimulating activities? 2) What are the biochemical components required for steroid-regulated transcription? 3) What factors determine whether a gene is or is not steroid responsive in a given cell (tissue) type? 4) How do steroid hormones alter the developmental pattern of gene expression in a well defined differentiating system? We will focus our attention on studies of the mouse glucocorticoid receptor and two specific genes that are regulated by glucocorticoids; one is the rat gene encoding the liver protein alpha-1 acid glycoprotein and the second is a mouse gene encoding a developmentally regulated cytochrome P450. It is our intent to eventually describe in detailed molecular terms the nature of the interactions between the glucocorticoid receptor and the transcriptional machinery of the cell. The approaches we are taking should help not only in elucidating the molecular mechanisms of steroid hormone action but may provide additional insights into the basis for tissue-specific gene expression and the process of cell differentiation.

几乎所有哺乳动物细胞都含有一种或多种受体 类固醇激素的类别。 这些高亲和力受体蛋白 最有可能介导类固醇对细胞功能的主要影响 通过与目标中（或附近）的特定 DNA 序列相互作用 基因。 这样做后，类固醇受体复合物会刺激 转录起始从而导致产量增加 特定 RNA。 复杂调控机制的研究 在高等真核生物中，由于以下困难而受到阻碍 通过基因和巨大的力量来操纵这些生物体 哺乳动物基因组。 因此我们转向简化 组织培养系统和分离基因来研究分子 类固醇激素调节基因表达的机制。 我们将使用遗传生化和分子方法 研究以下问题： 1) 结构是什么？ 糖皮质激素受体以及蛋白质的哪些部分 激素结合、DNA 结合和转录所需 刺激活动？ 2）生化成分是什么 类固醇调节转录所需的？ 3）哪些因素 确定一个基因在给定的情况下是否对类固醇有反应 细胞（组织）类型？ 4) 类固醇激素如何改变 基因表达的发育模式有明确的定义 差异化系统？ 我们将重点关注小鼠糖皮质激素的研究 受体和两个受调节的特定基因 糖皮质激素；一是编码肝脏蛋白的大鼠基因 α-1酸性糖蛋白，第二个是编码小鼠基因 发育调节的细胞色素 P450。 我们的目的是 最终以详细的分子术语描述了 糖皮质激素受体与糖皮质激素受体之间的相互作用 细胞的转录机制。 我们采取的方法 服用不仅有助于阐明分子 类固醇激素作用机制，但可能提供额外的 深入了解组织特异性基因表达的基础和 细胞分化的过程。