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.

我的近期目标是发展与以下领域同等的研究背景 我已经致力于临床工作。 因此，我决定追求 在凯斯西储大学跟随 John Nilson 博士获得博士学位 大学（CWRU）。 我的长期职业目标是结合我的才能 在基础科学和临床医学领域担任教职 兽医教学机构。 尼尔森博士实验室的程序 CWRU 的环境提供了获得任何所需的非凡机会 资源。 我的研究事业发展将在此得到培养 能够参加大量结构化会议和培训的环境 机会。 在这里我将学习如何批判性地评估数据 将使我能够计划合乎逻辑的新实验。 另外，我会 接受宝贵的写作简洁和有意义的培训 科学研究论文和资助提案。这些成就是 对于我争取成为独立人士的最终目标至关重要 研究员。 这笔赠款解决了雄激素的总体工作假设 雌激素受体通过多种方式抑制促黄体素的表达 促性腺激素中的蛋白质-蛋白质相互作用。有证据表明 促性腺激素中的 AR 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身份仍然存在 未知。 在目标 1 中，酵母双杂交方法将用于 筛选共阻遏蛋白的定向和随机方式。 在 此外，我们假设至少有一个磷酸化残基 AR 是与共阻遏蛋白成功相互作用所必需的。 我们打算通过瞬时转染来测试目标 2 中的这一假设 带有α亚基启动子的AR磷酸化突变体 促性腺激素细胞。 最后，我们计划利用所获得的知识 α亚基抑制机制使我们能够解锁 LHbeta 调节背后的谜团。 在目标 3 中，我们建议设计 转基因小鼠以确定其位点和机制 LHbeta 亚基基因的类固醇生成调节。