Biochemical, structural and molecular dissection of androgen receptor transcriptional activity

项目摘要和摘要 雄激素受体(AR)是主要的转录因子，负责发育和维持 男性的性别表型。AR活性异常与多种疾病有关，包括但不限于 转移性前列腺癌、雄激素性脱发和肯尼迪病。基于细胞的研究报告称 AR通过许多过程进行调节，包括1)雄激素刺激；2)抗雄激素抑制；3) 其结构域之间的分子内接触；4)与各种蛋白质辅活化子和辅阻遏子的结合。 尽管AR在疾病病因学中很重要，但AR调节的机制细节在很大程度上 未知，因为历史上很难分离出纯的、活跃的多域AR变体。 这项研究将使用生化和互补的结构方法--包括电子和原子 力显微镜和X射线结晶学-阐明AR激活的分子决定因素 识别对分子内和辅因子介导的刺激(目标1)和AR重要的表面 通过抗雄激素结合的抑制和自我抑制的自我调节模式(目标3)。分子细节 生化和结构研究揭示的结果将在生物相关的AR依赖细胞中得到验证 用于确定这些相互作用对细胞形态、生长、AR信号和 AR环路(目标2)。这些研究的发现将为二聚化模式提供新的见解， AR和其他I型核激素受体的分子内调节和DNA结合，包括 孕酮、糖皮质激素和盐皮质激素受体，并可促进潜在的新型设计 AR-靶向药物。这项研究与NIGMS生成基础知识的目标是一致的 这将对人类疾病和健康产生影响。 所描述的全面培训计划将使调查员能够实现她的跑步职业目标 一个独立的学术实验室，致力于研究AR转录调控的基本模式 在其活跃、过度活跃和抑制状态的上下文中，并将提供生化、结构和 对其在疾病中异常活动的后果的功能性洞察。作为顶级前列腺癌的联合导师 纪念斯隆·凯特琳癌症中心和洛克菲勒癌症和结构生物学实验室 在大学期间，她将接受不同的前列腺生物学和电子显微镜培训，这将扩大 研究人员的生物学理解和技术技能，将使HER实现重大创新 未来的研究。她将通过马赛克课程获得的领导力、网络和沟通技能 UE5将促进她向独立的过渡，并使她具备增强科学工作队伍多样性的能力。

PROJECT SUMMARY AND ABSTRACT The androgen receptor (AR) is the master transcription factor responsible for development and maintenance of the male sex phenotype. Aberrant AR activity is associated with multiple disorders, including but not limited to metastatic prostate cancer, androgenetic alopecia, and Kennedy's disease. Cell-based studies have reported AR is regulated through numerous processes, including 1) androgen stimulation; 2) anti-androgen inhibition; 3) intramolecular contacts among its domains; 4) association with various protein coactivators and corepressors. Despite its importance in disease etiology, the mechanistic details underlying AR regulation are largely unknown due to historical difficulty isolating pure, active multidomain AR variants. This research will employ biochemical and complementary structural methods – including electron and atomic force microscopy, and x-ray crystallography – to illuminate the molecular determinants of AR activation through identification of surfaces important for intramolecular and cofactor-mediated stimulation (Aim 1), and for AR inhibition by anti-androgen binding and self-regulatory modes of autoinhibition (Aim 3). The molecular details revealed by the biochemical and structural studies will be validated in biologically relevant AR-dependent cell lines to determine the functional outcomes of these interactions on cell morphology, growth, AR signaling and the AR cistrome (Aim 2). The findings from these studies will provide novel insight for modes of dimerization, intramolecular regulation, and DNA binding for AR as well as other type I nuclear hormone receptors, including the progesterone, glucocorticoid, and mineralocorticoid receptors, and can facilitate potential design of novel AR-targeting drugs. This research is in line with the goals of the NIGMS of generating fundamental knowledge that will bear implication for human disease and health. The comprehensive training plan described will enable the investigator to achieve her career goals of running an independent academic laboratory dedicated to studying fundamental modes of AR transcriptional regulation in the context of its active, hyperactivated, and inhibited states, and will provide biochemical, structural, and functional insight to the consequences of its aberrant activity in disease. As a joint mentee in top prostate cancer and structural biology laboratories at Memorial Sloan Kettering Cancer Center and the Rockefeller University, she will receive diverse training in prostate biology and electron microscopy that will expand the investigator's biological understanding and technical repertoire that will enable significant innovation in her future research. The leadership, networking, and communication skills she will acquire through the MOSAIC UE5 will facilitate her transition to independence, and equip her to enhance diversity in the scientific workforce.