MOLECULAR BIOLOGY OF STEROID HORMONE ACTION

类固醇激素通过特定的受体发挥作用， 几乎所有动物细胞的基因表达。 我们建议研究 组织培养中糖皮质激素作用的分子生物学 来源于小鼠乳腺癌和大鼠肝癌细胞的系统。 的 小鼠乳腺肿瘤病毒（MMTV）RNA的产生受到以下刺激： 乳腺细胞和MMTV感染的大鼠肝癌中糖皮质激素的变化 细胞 使用各种分子技术（包括核酸 杂交和RNA测序），我们将研究生物化学过程， 刺激病毒RNA合成;特别是，我们将尝试 鉴定病毒RNA合成起始的启动子位点。 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的转录系统， 体外 利用免疫选择技术， 肝癌细胞将发展，其中一种或多种成分涉及 是有缺陷的 这些突变体将被用来识别 参与激素作用的分子成分， 控制相关的生化观察与事件发生在 vivo. 这些方法不仅可以阐明分子机制， 类固醇激素的作用，但也提供了一些进一步的见解， 真核基因调控一般和事件导致病毒诱导 肿瘤发生

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.