STEROIDS REPRESS LH THROUGH PROTEIN/PROTEIN INTERACTIONS

类固醇通过蛋白质/蛋白质相互作用抑制 LH

• 批准号: 6498097
• 负责人: Joan S Jorgensen
• 金额: $ 4.07万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2002-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6498097
• 关键词: DNA binding protein; androgen receptor; estrogen receptors; genetic library; genetic regulation; genetically modified animals; hormone regulation /control mechanism; laboratory mouse; luteinizing hormone; phosphorylation; protein protein interaction; receptor expression; steroid hormone receptor; transfection; yeast two hybrid system

My immediate goal is to develop an equal background in research to what I had already devoted to clinical work. Therefore, I decided to pursue a doctoral degree with Dr. John Nilson at Case Western Reserve University (CWRU). My long term career goals are to combine my talents in basic science and clinical medicine in a faculty position at a veterinary teaching institution. The program in Dr. Nilson's laboratory and the environment at CWRU provide extraordinary access to any required resources. My research career development will be nurtured in this environment with the access to numerous structured meetings and training opportunities. Here I will learn how to critically evaluate data which will allow me to plan logical new experiments. In addition, I will receive invaluable training in writing succinct and meaningful scientific research papers and grant proposals. These achievements are critical in striving for my ultimate goal of becoming an independent researcher. This grant addresses the overall working hypothesis that the androgen and estrogen receptors repress lutropin expression through multiple protein-protein interactions in gonadotropes. Evidence indicates that AR in gonadotropes suppresses the alpha subunit promoter through protein-protein interactions that involve two distinct regulatory elements. However the identity of these critical proteins remains unknown. In Aim 1, the yeast two-hybrid approach will be used in both a directed and random fashion to screen for co-repressor proteins. In addition, we postulate that at least one phosphorylated residue of the AR is required for successful interaction with the co-repressor protein. We intend to test this hypothesis in Aim 2 by transiently transfecting AR phosphorylation mutants with the alpha subunit promoter in gonadotrope cells. Finally, we plan to exploit the knowledge gained on the alpha subunit suppression mechanism to enable us to unlock the mystery behind LHbeta regulation. In Aim 3, we propose to design transgenic mice in order to define the site and mechanism of steroidogenic regulation on the LHbeta subunit gene.

我的直接目标是在研究方面建立一个与 我已经投入到临床工作中了。因此，我决定继续 凯斯西储大学约翰·尼尔森博士的博士学位 大学(CWRU)。我的长期职业目标是把我的才能结合起来 在基础科学和临床医学方面担任教职 兽医教学机构。尼尔森博士实验室里的程序 CWRU的环境提供了非凡的访问权限 资源。我的研究生涯将在这里得到培育 能够参加大量有组织的会议和培训的环境 机遇。在这里，我将学习如何批判性地评估数据 会让我计划合乎逻辑的新实验。另外，我会 接受无价之宝的写作训练，简洁而有意义 科研论文和资助建议。这些成就是 在为我成为一个独立的人的最终目标而奋斗时至关重要 研究员。 这项拨款解决了总体上的工作假设，即雄激素 雌激素受体通过多种途径抑制促黄体生成素的表达 促性腺激素中的蛋白质-蛋白质相互作用。有证据表明， 促性腺激素受体通过抑制α亚基启动子 蛋白质-蛋白质相互作用涉及两种不同的调节 元素。然而，这些关键蛋白质的特性仍然存在 未知。在目标1中，酵母双杂交方法将在这两种方法中使用 一种定向和随机的方式来筛选共抑制蛋白。在……里面 此外，我们假设至少有一个磷酸化残基 AR是与共抑制蛋白成功相互作用所必需的。 我们打算通过瞬时转染法在目标2中验证这一假设。 含α亚基启动子的AR磷酸化突变体 促性腺激素细胞。最后，我们计划利用从 阿尔法亚单位抑制机制使我们能够解锁 LHbeta调控背后的奥秘。在目标3中，我们建议设计 转基因小鼠，以确定其作用部位和机制 LHbeta亚基基因的类固醇合成调控。