GENE REGULATION BY STEROID RECEPTOR PROTEINS

糖皮质激素受体，与其同源类固醇结合后 配体，结合到特定的DNA位点，从而激活受体依赖性 转录增强子元件，其反过来刺激从 附近的促销员 本研究的长期目标是确定 类固醇受体调节基因的分子机制 表达式，以定义这些机制如何与其他模式相关 的基因控制，并推断途径，协调网络， 受调控的基因可能会出现并进化。 在本申请中， 重组DNA方法将用于构建一系列广泛的 在DNA的离散区域中的突变，这些突变特异性地与纯 糖皮质激素受体，并在体内有活性， 受体依赖性增强子。 连同完整的，切割的，和突变的 受体蛋白，我们将在核苷酸水平上定义 特异性受体的特征：DNA相互作用， 这些相互作用对DNA和染色质构象在体内和 体外，以及结合和模板结构的精确关系 转录增强机制的改变。 此外，本发明还提供了一种方法， 将寻找激素调节所必需的非受体因子， 以生物化学、细胞生物学和体细胞遗传学为特征 程序. 了解调节基因表达的机制是 人类疾病和发育缺陷问题的核心;类固醇 特别是激素，可以调节广泛的发育和 生理现象，并且是广泛使用的药理学试剂，用于 免疫抑制和癌症化疗。 （丁）

Glucocorticoid receptors, upon association with their cognate steroid ligands, bind to specific DNA sites, thereby activating receptor-dependent transcriptional enhancer elements that in turn stimulate initiation from nearby promoters. The long-term objectives of this study are to determine the molecular mechanisms by which steroid receptors regulate gene expression, to define how these mechanisms might be related to other modes of gene control, and to infer pathways by which coordinated networks of regulated genes might arise and evolve. In the present application, recombinant DNA methods will be used to construct an extensive series of mutations in discrete regions of DNA that are bound specifically by pure glucocorticoid receptor in vitro and are active in vivo as receptor-dependent enhancers. Together with intact, cleaved, and mutant receptor proteins, we will define at the nucleotide level the characteristics of specific receptor: DNA interactions, the consequences of those interactions on DNA and chromatin conformation in vivo and in vitro, and the precise relationships of the binding and template structure alterations to the mechanism of transcriptional enhancement. In addition, nonreceptor factors essential for hormonal regulation will be sought and characterized by biochemical, cell biological, and somatic cell genetic procedures. Understanding the mechanisms that modulate gene expression is central to the problems of human disease and developmental defects; steroid hormones, in particular, regulate a broad spectrum of developmental and physiological phenomena, and are widely used pharmacological agents for immunosuppression and cancer chemotherapy. (D)