HARC: HIV accessory and regulatory complexes

HARC中心：HIV附件和调控复合体 总体总结 HARC中心是一个跨学科的项目，旨在提高我们对相互作用的理解 HIV附件和调节蛋白与宿主细胞系统之间的关系，最终目标是扩大 艾滋病毒/艾滋病的治疗目标和治疗方式。HARC中心将专注于 HIV-1辅助蛋白和调节蛋白结构的测定，重点是Tat，Vif和Rev， 与其细胞伙伴的关系复杂，到目前为止，这些伙伴还不是艾滋病毒治疗方式的目标。一个 更好地了解病毒-宿主复合体的功能和机制可能会揭示新的 干预的治疗策略，包括宿主定向治疗的策略，这些策略可能会逃脱 目前药物方案的局限性，靶向艾滋病毒酶的突变可能会降低药物疗效。 HARC中心将使用集成的“系统到宿主”来确定这些病毒-宿主复合体的结构。 结构“平台，包括(1)通过功能蛋白质组学和遗传学的新方法发现 在HARC中心开发，(2)通过突破性的CRISPR方法在原代T细胞中进行验证 以及靶向生化和功能分析，以及(3)利用合成的 在HARC中心以前的迭代中开发的创新结构技术，以解决大型、 我们研究的是柔性的、多相的，有时还与膜相关的系统。 拟议的研究项目围绕服务于中心目标的三个主题展开。我们将探索 HIV如何抑制宿主限制因子(主题1)，研究调节HIV转录和潜伏期的因素 (主题2)和调查三个项目的病毒-宿主进化(主题3)：VIF的结构和进化 和APOBEC3(项目1)，艾滋病毒转录和潜伏期的调节(项目2)，以及遗传学和 进化(项目3)。HARC中心的项目由4个涵盖蛋白质组的技术核心支持 研究病毒的方法(核心1-蛋白质组学)、CRISPR筛查和原代细胞内源性标记 宿主功能(核心2-遗传学)，使用冷冻电子显微镜的结构生物学，X射线筛查和 确定艾滋病毒-宿主复合体结构的抗体技术(核心3--结构生物学)， 以及病毒-宿主复合体的综合建模(核心4-计算)。总体目标、进展和 行政以及外联活动和通信将由行政核心监督 (核心5)。发展核心(核心6)将为年轻的调查人员和HARC提供培训机会 中心成员，并授予合作机会基金，以加强中心的整体研究主题。

THE HARC CENTER: HIV ACCESSORY AND REGULATORY COMPLEXES OVERALL SUMMARY The HARC Center is an interdisciplinary program that aims to improve our understanding of the interactions between HIV accessory and regulatory proteins and host cellular systems, with the ultimate goal to expand on therapeutic targets and treatment modalities for HIV/AIDS. The HARC Center will focus its efforts on the determination of structures of the accessory and regulatory proteins of HIV-1, with a focus on Tat, Vif and Rev, in complex with their cellular partners, which have not been targets of HIV therapeutic modalities until now. A better molecular understanding of the functions and mechanisms of virus-host complexes may reveal new therapeutic strategies for intervention, including strategies of host-directed therapies, which may escape the limitations of current drug regimens where mutations in the targeted HIV enzymes can diminish drug efficacy. The HARC Center will determine the structures of these virus-host complexes using an integrated “Systems-to- Structure” platform that includes (1) Discovery, through novel methods of functional proteomics and genetics being developed in the HARC Center, (2) Validation through breakthrough CRISPR methods in primary T cells as well as targeted biochemical and functional assays, and (3) Structure Determination using a synthesis of innovative structural techniques developed in previous iterations of the HARC Center to address the large, flexible, heterogeneous and sometimes membrane-associated systems we study. The proposed Research Projects are centered around three themes that serve the Center’s goal. We will explore how HIV inhibits Host restriction factors (Theme 1), study the factors regulating HIV transcription and latency (Theme 2) and investigate virus-host Evolution (Theme 3) across three projects: Structure and evolution of Vif and APOBEC3 (Project 1), Regulation of HIV transcription and latency (Project 2), and Genetics and evolution (Project 3). The HARC Center projects are supported by 4 technology cores covering proteomic approaches (Core 1 - Proteomics), CRISPR screens and endogenous tagging in primary cells to study virus- host function (Core 2 - Genetics), structural biology using cryo-Electron Microscopy, X-ray screening and antibody technologies to determine the structures of HIV virus-host complexes (Core 3 - Structural Biology), and integrative modeling of virus-host complexes (Core 4 - Computational). The overall goals, progress, and administration as well as outreach activities and communications will be overseen by the Administrative Core (Core 5). The Developmental Core (Core 6) will provide training opportunities to young investigators and HARC Center members and award the Collaborative Opportunity Fund to enhance the Center’s overall research theme.