MOLECULAR BIOLOGY OF STEROID HORMONE ACTION

类固醇激素作用的分子生物学

• 批准号: 3273330
• 负责人: GORDON M RINGOLD
• 金额: $ 24.76万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-12-01 至 1986-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3273330
• 关键词: Escherichia coli; clone cells; endonuclease; eukaryote; flow cytometry; gene expression; genetic manipulation; genetic recombination; genetic transcription; glucocorticoids; hepatocellular carcinoma; hormone regulation /control mechanism; immunochemistry; liver; messenger RNA; microorganism genetics; molecular cloning; monoclonal antibody; mouse mammary tumor virus; nucleic acid sequence; radiotracer; spectrometry; steroid hormone; temperature sensitive mutant; vaccinia virus; viral carcinogenesis; virus RNA

Steroid hormones function via specific receptors to exert dramatic effects on gene expression in virtually all animal cells. We propose to study the molecular biology of glucocorticoid action in well-defined tissue culture systems derived from mouse mammary carcinoma and rat hepatoma cells. The production of mouse mammary tumor virus (MMTV) RNA is stimulated by glucocorticoid hormones in the mammary cells and in MMTV-infected rat hepatoma cells. Using a variety of molecular techniques (including nucleic acid hybridization and RNA sequencing) we will study the biochemical processes by which viral RNA synthesis is stimulated; in particular, we will attempt to identify the promotor site(s) at which viral RNA synthesis is initiated. Transcription of viral genes will be studied in isolated nuclei and chromatin in order to eventually reconstruct hormone dependent transcription systems in vitro. Using immunological selection techniques, genetic mutants of the rat hepatoma cells will be developed in which one or more of the components involved in the steroid response is defective. These mutants would be used to identify the molecular components involved in hormone action and to serve as genetic controls in correlating biochemical observations with the events which occur in vivo. These approaches may serve not only to elucidate the molecular mechanisms of steroid hormone action but also to provide some further insights into eukaryotic gene regulation in general and the events leading to virally induced tumorigenesis.

类固醇激素通过特定受体发挥作用，对身体产生巨大影响 几乎所有动物细胞中的基因表达。 我们建议研究 明确组织培养中糖皮质激素作用的分子生物学 源自小鼠乳腺癌和大鼠肝癌细胞的系统。 这 小鼠乳腺肿瘤病毒 (MMTV) RNA 的产生受到以下因素的刺激 乳腺细胞和 MMTV 感染的大鼠肝癌中的糖皮质激素 细胞。 使用各种分子技术（包括核酸 杂交和 RNA 测序）我们将通过以下方式研究生化过程 哪种病毒RNA合成受到刺激；特别是，我们将尝试 识别启动病毒 RNA 合成的启动子位点。 将在分离的细胞核和染色质中研究病毒基因的转录 为了最终重建激素依赖性转录系统 体外。 使用免疫选择技术，大鼠的基因突变体 将开发出含有一种或多种成分的肝癌细胞 类固醇反应有缺陷。 这些突变体将被用来识别 参与激素作用并作为遗传物质的分子成分 将生化观察结果与发生的事件相关联的控制 体内。 这些方法不仅可以阐明分子机制 类固醇激素作用的同时也提供了一些进一步的见解 一般真核基因调控以及导致病毒诱导的事件 肿瘤发生。