MULTIHORMONAL REGULATION OF GENE EXPRESSION

大多数真核基因受到多因子调控， 发育、激素、组织特异性和/或环境线索。 的 因此，从基因中产生的蛋白质的净含量必须反映这些蛋白质是如何被合成的。 信号集成。 我们的长期目标是了解这些复杂的 调节事件被协调以引发适当水平的基因表达， 表情 鸡卵清蛋白（Ov）基因是一个很好的模型 研究基因表达的多因素调控，因为它是 在转录水平上由四类类固醇调节 激素，胰岛素和其他积极和消极的调制器，在一个 严格的组织特异性方式。 该提案的主要重点是 了解类固醇激素如何激活Ov基因。 固有 对这个问题的理解是分析类固醇如何起作用 协同地激活迟发反应基因。 由于大多数基因诱导 类固醇是这个类的成员，这是必不可少的， 理解激活之间的中间事件 早期基因的类固醇受体和随后的激活 晚期基因 该补助金的具体目标是我。识别和 描述了类固醇反应性早期反应蛋白， Ov基因，II. 研究类固醇激素的协同作用 对Ov基因转录的影响，和III. 审查《公约》的作用 在调节Ov基因表达中的负调控元件。 到 为此，基因调控蛋白将被克隆和表征 与Ov基因中的主要调控元件结合， 类固醇依赖性调节元件（SDRE）和负调节元件 元素（NRE）。 用接头扫描突变的转染实验 通过SDRE和NRE将被用来定义特定的角色， 每个蛋白质结合位点。 基因组足迹将有助于 在确定两种类固醇如何实现协同作用以及SDRE如何 和NRE充当单个功能实体。 由于类固醇激素一直是 与某些癌症的病因学有关，更好地了解 由类固醇引发的分子事件只能改善治疗， 预防这些恶性肿瘤。 此外，这些知识可能 提供关于开发更好的避孕药具的见解， 治疗生殖障碍

Most eucaryotic genes are subject to multifactorial regulation by developmental, hormonal, tissue-specific, and/or environmental cues. The net amount of protein made from a gene must therefore reflect how these signals integrate. Our long term goal is to understand how these complex regulatory events are coordinated to elicit appropriate levels of gene expression. The chicken ovalbumin (Ov) gene serves as an excellent model for studying multifactorial regulation of gene expression because it is regulated at the transcriptional level by four classes of steroid hormones, by insulin, and by other positive and negative modulators in a strictly tissue-specific manner. The major focus of this proposal is to understand how steroid hormones activate the Ov gene. Intrinsic to the comprehension of this problem is an analysis of how steroids function synergistically to activate late response genes. As most genes induced by steroids are members of this class, it is essential that an understanding be achieved of the intermediary events between activation of the early genes by steroid receptors and the subsequent activation of the late genes. The Specific Aims of this grant are to I. Identify and characterize the steroid-responsive early response proteins that regulate the Ov gene, II. Investigate the synergistic effects of steroid hormones on the transcription of the Ov gene, and III. Examine the role of the negative regulatory element in modulating expression of the Ov gene. To this end, the gene regulatory proteins will be cloned and characterized that bind to the major regulatory elements in the Ov gene, the steroid-dependent regulatory element (SDRE) and the negative regulatory element (NRE). Transfection experiments with linker scanning mutations through the SDRE and NRE will be employed to define specific roles for each protein binding site. Genomic footprinting will be done to assist in determining how synergism is achieved by two steroids and how the SDRE and NRE act as a single functional entity. As steroid hormones have been implicated in the etiology of some cancers, a better understanding of the molecular events triggered by steroids can only improve the treatment and prevention of these malignancies. Furthermore, such knowledge may provide insights into the development of better contraceptives and aid in the treatment of reproductive disorders.