HARC Center: HIV Accessory and Regulatory Complexes

HARC 中心：HIV 辅助和调节复合体

• 批准号: 7933127
• 负责人: ALAN D FRANKEL
• 金额: $ 60万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7933127
• 关键词: HARC; Center; HIV; Accessory; Regulatory

Throughout the HIV life cycle, viral-host complexes play an integral role in the biology of the virus. Assemblies range from binary protein-protein and protein-nucleic acid complexes to much larger multicomponent complexes. High-resolution structures of these complexes offer the potential to provide templates for intervention strategies in the treatment of AIDS, but structures have so far been solved in only a few cases. Obtaining such structural information has been limited in part because several of the HIV proteins are poorly behaved biochemically and often are unstructured in isolation. These characteristics may reflect an inherent property of the proteins to adopt different conformations when bound transiently to differen cellular partners throughout the viral life cycle. This project, called the HARC Center (HIV Accessory and Regulatory Complexes), focuses on five key HIV proteins that perform important regulatory and accessory functions - Integrase (IN), Tat, Rev, Vif, and Nef- and their interacting viral and cellular protein partners. Our primary aim is to achieve a comprehensive structural picture of HIV-host cell interactions formed during early phases of the viral life cycle, particularly those that regulate gene expression, cell signaling, or cell biology. Several of these viral and host proteins undergo significant conformational changes when complexed to their nucleic acid or protein partners, and understanding the molecular details of this dynamic behavior is an important structural goal. Our focus on regulatory complexes will provide new insights into the biology and pathogenesis of HIV and define new therapeutic targets to combat HIV infection. The HARC Center will harness protein expression and co-expression strategies to characterize functional complexes using a comprehensive battery of structural methods, including mass spectrometry, xray crystallography, NMR, and cryo-EM. To overcome the obstacles that have limited progress to date, we propose substantial efforts for technological development. Essential new methods will be explored to optimize expression-to-crystallization, analyze protein dynamics using crystallographic and NMR data, and determine structures of small and heterogeneous complexes by cryo-EM. Strong outreach and collaborative components will draw investigators from outside the Center, particularly those with expertise in HIV mechanisms, protein biochemistry, and virology.

在整个艾滋病毒生命周期中，病毒-宿主复合物在病毒生物学中发挥着不可或缺的作用。 组装范围从二元蛋白质-蛋白质和蛋白质-核酸复合物到更大的 多组分复合物。这些复合物的高分辨率结构有可能提供 艾滋病治疗干预策略的模板，但迄今为止结构已得到解决 只有少数情况。获得此类结构信息的部分原因是由于一些艾滋病毒 蛋白质的生化行为很差，并且通常在分离时是非结构化的。这些特征可能 反映了蛋白质的固有特性，当瞬时结合到不同的物质上时，会采用不同的构象 整个病毒生命周期的细胞伙伴。 该项目称为 HARC 中心（HIV 辅助和调节复合体），重点关注五个 执行重要调节和辅助功能的关键 HIV 蛋白 - Integrase (IN)、Tat、Rev、Vif、 和 Nef- 以及它们相互作用的病毒和细胞蛋白伙伴。我们的首要目标是实现 病毒生命早期阶段形成的 HIV-宿主细胞相互作用的全面结构图 循环，特别是那些调节基因表达、细胞信号传导或细胞生物学的循环。其中有几个病毒 当与核酸或宿主蛋白复合时，宿主蛋白会发生显着的构象变化。 蛋白质伙伴，了解这种动态行为的分子细节是一个重要的结构 目标。我们对监管复合物的关注将为艾滋病毒的生物学和发病机制提供新的见解 并确定新的治疗靶点来对抗艾滋病毒感染。 HARC 中心将利用蛋白质表达和共表达策略来表征 使用全面的结构方法（包括质谱、X 射线）构建功能复合物 晶体学、NMR 和冷冻电镜。为了克服迄今为止进展有限的障碍，我们 提出大力推进科技发展的建议。将探索重要的新方法 优化表达到结晶，使用晶体学和 NMR 数据分析蛋白质动力学，以及 通过冷冻电镜确定小且异质复合物的结构。强大的外展和协作能力 组成部分将吸引中心外部的研究人员，特别是那些具有艾滋病毒专业知识的研究人员 机制、蛋白质生物化学和病毒学。