HARC Center: HIV Accessory and Regulatory Complexes

描述（由申请方提供）：在整个HIV生命周期中，病毒-宿主复合物在病毒生物学中起着不可或缺的作用。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为治疗艾滋病的干预策略提供模板的可能性，但迄今为止仅在少数情况下解决了结构。获得这样的结构信息是有限的，部分原因是一些HIV蛋白质的生物化学行为很差，并且通常是孤立的非结构化的。这些特征可能反映了蛋白质在整个病毒生命周期中瞬时结合不同细胞伴侣时采用不同构象的固有特性。该项目被称为HARC中心（HIV辅助和调节复合物），重点关注执行重要调节和辅助功能的五种关键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质伴侣。我们的主要目标是实现一个全面的结构图的HIV-宿主细胞相互作用形成的病毒生命周期的早期阶段，特别是那些调节基因表达，细胞信号传导，或细胞生物学。这些病毒和宿主蛋白质中的一些在与其核酸或蛋白质伴侣复合时经历显著的构象变化，并且理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以对抗HIV感染。HARC中心将利用蛋白质表达和共表达策略，使用一系列全面的结构方法来表征功能复合物，包括质谱法，X射线晶体学，NMR和cryo-EM。为了克服迄今为止限制进展的障碍，我们建议为技术发展作出重大努力。将探索必要的新方法来优化表达结晶，使用晶体学和NMR数据分析蛋白质动力学，并通过cryo-EM确定小的和异质的复合物的结构。强有力的外联和协作部分将吸引来自中心以外的研究人员，特别是那些在艾滋病毒机制，蛋白质生物化学和病毒学方面具有专业知识的人员。 整体

DESCRIPTION (provided by applicant): 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. OVERALL