HARC Center: HIV Accessory and Regulatory Complexes

HARC 中心：HIV 辅助和调节复合体

• 批准号: 8119501
• 负责人: ALAN D FRANKEL
• 金额: $ 336.59万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-27 至 2012-09-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8119501
• 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 different 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, x- ray 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的生物学中起着不可或缺的作用。组件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络合物。这些复合物的高分辨率结构提供了为艾滋病治疗的干预策略提供模板的潜力，但是迄今仅在少数情况下解决了结构。获得此类结构信息的部分受到限制，部分是因为几种HIV蛋白在生化上表现不佳，并且通常是孤立的。这些特征可能反映了蛋白质在整个病毒生命周期中暂时与不同的细胞伴侣瞬时结合时采用不同构象的固有特性。 该项目称为HARC中心（HIV附件和调节络合物），重点介绍了五种关键的HIV蛋白，这些蛋白质具有重要的调节和辅助功能 - Integrase（In），TAT，REV，VIF和NEF-及其相互作用的病毒和细胞蛋白质伴侣。我们的主要目的是实现病毒生命周期早期阶段中形成的HIV宿主细胞相互作用的综合结构图，尤其是那些调节基因表达，细胞信号传导或细胞生物学的结构图。这些病毒和宿主蛋白中的几个在与核酸或蛋白质伴侣复合时会发生显着构象变化，并且了解这种动态行为的分子细节是一个重要的结构目标。我们对调节络合物的关注将为艾滋病毒的生物学和发病机理提供新的见解，并定义新的治疗靶标，以打击HIV感染。 HARC中心将利用全面的结构方法（包括质谱，X-Ray晶体学，NMR和Cryo-EM）来利用蛋白质表达和共表达策略来表征功能复合物。为了克服迄今为止进步有限的障碍，我们为技术发展做出了重大努力。将探索基本的新方法，以优化表达到结晶，使用晶体学和NMR数据分析蛋白质动力学，并确定Cryo-EM的小型和异构复合物的结构。强大的外展和协作组件将吸引中心外的研究人员，尤其是那些在HIV机制，蛋白质生物化学和病毒学方面具有专业知识的研究者。