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-宿主复合体在病毒生物学中发挥着不可或缺的作用。 组装的范围从二元蛋白质-蛋白质和蛋白质-核酸复合体到更大的 多组分络合物。这些络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，但到目前为止，结构已在#年得到解决 只有几个案子。获得这种结构信息受到限制，部分原因是几种艾滋病毒 蛋白质在生物化学方面表现不佳，通常在分离时是无结构的。这些特征可能 反映蛋白质在瞬间结合到不同时采用不同构象的固有属性 在病毒的整个生命周期中都有细胞伙伴。 这个项目被称为HARC中心(艾滋病毒附件和管理复合体)，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)、TAT、REV、VIF、 和Nef-以及它们相互作用的病毒和细胞蛋白伙伴。我们的主要目标是实现 在病毒生命的早期阶段形成的艾滋病毒-宿主细胞相互作用的全面结构图 周期，尤其是那些调节基因表达、细胞信号或细胞生物学的基因。其中几个像病毒一样 当宿主蛋白与它们的核酸或 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 进球。我们对调控复合体的关注将为HIV的生物学和发病机制提供新的见解 并确定新的治疗目标，以抗击艾滋病毒感染。 HARC中心将利用蛋白质表达和共表达策略来表征 功能络合物采用全面的电池结构方法，包括质谱学、X-射线衍射法 结晶学、核磁共振和低温电子显微镜。为了克服迄今限制进展的障碍，我们 建议在技术发展方面做出实质性努力。将探索必要的新方法来 优化表达到结晶，使用结晶学和核磁共振数据分析蛋白质动力学，以及 用低温电子显微镜测定小分子和多相配合物的结构。强大的外延和协作性 各部分将吸引中心以外的调查人员，特别是那些具有艾滋病毒专业知识的调查人员 机制、蛋白质生物化学和病毒学。