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发展平等的背景 我已经致力于临床工作。 因此，我决定追求 与约翰·尼尔森（John Nilson）博士的博士学位 大学（CWRU）。 我的长期职业目标是结合我的才华 在教师的基础科学和临床医学 兽医教学机构。 尼尔森博士实验室的计划 CWRU的环境提供了对任何必需的任何要求的非凡访问 资源。 我的研究职业发展将在此培养 可以访问众多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评估数据 将允许我计划逻辑新实验。 此外，我会 接受书面简洁而有意义的书面培训 科学研究论文和授予建议。这些成就是 追求我成为独立的最终目标至关重要 研究员。 该赠款解决了雄激素的总体工作假设 和雌激素受体通过多种 促性腺激素中的蛋白质蛋白质相互作用。证据表明 促性腺导体中的AR通过 蛋白质 - 蛋白质相互作用，涉及两个不同的调节 元素。 但是这些关键蛋白的身份仍然存在 未知。 在AIM 1中，酵母双杂交方法将在两者中使用 一种定向和随机的时尚，用于筛选共抑制蛋白。 在 此外，我们假设至少一个磷酸化的残基 与共抑制蛋白的成功相互作用是必需的。 我们打算通过瞬时转染在AIM 2中检验这一假设 用α亚基启动子在AR磷酸化突变体中 促性腺细胞。 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管制的神秘。 在AIM 3中，我们建议设计 转基因小鼠以定义位点和机制 LHBETA亚基基因上的类固醇调节。