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HIV生命周期中，病毒-宿主复合物在病毒的生物学中发挥着不可或缺的作用。 组装体的范围从二元蛋白质-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提供了提供 艾滋病治疗干预战略的模板，但迄今为止， 只有几个案例。获得这样的结构信息是有限的，部分原因是一些艾滋病毒 蛋白质在生物化学上表现较差，并且在分离时通常是非结构化的。这些特性可以 反映了蛋白质的固有性质，即当与蛋白质瞬时结合时， 在整个病毒生命周期中的细胞伴侣。 这个项目，称为HARC中心（艾滋病毒配件和监管复合体），重点是五个 执行重要调节和辅助功能的关键HIV蛋白-整合酶（IN），达特，Rev，Vif， 和Nef-以及它们相互作用的病毒和细胞蛋白伴侣。我们的主要目标是实现 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 细胞周期，特别是那些调节基因表达，细胞信号传导或细胞生物学。其中一些病毒 并且宿主蛋白质在与它们的核酸复合时经历显著的构象变化， 蛋白质伴侣，了解这种动态行为的分子细节是一个重要的结构 目标.我们对调节复合物的关注将为HIV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 功能性复合物，使用结构方法的综合电池，包括质谱，X射线 晶体学核磁共振和冷冻电镜为了克服迄今为止限制进展的障碍，我们 大力推进技术开发。将探索必要的新方法， 优化表达到结晶，使用晶体学和NMR数据分析蛋白质动力学， 通过冷冻电镜确定小的和异质复合物的结构。强有力的外联和协作 这些组成部分将吸引中心以外的调查人员，特别是那些在艾滋病毒方面有专长的人 机制、蛋白质生物化学和病毒学。