HARC: HIV accessory and regulatory complexes

HARC 中心：HIV 配件和调节复合体 总体总结 HARC 中心是一个跨学科项目，旨在提高我们对相互作用的理解 HIV辅助蛋白和调节蛋白与宿主细胞系统之间的关系，最终目标是扩展 艾滋病毒/艾滋病的治疗目标和治疗方式。 HARC 中心将重点致力于 确定 HIV-1 的辅助蛋白和调节蛋白的结构，重点是 Tat、Vif 和 Rev， 与它们的细胞伴侣形成复合体，而迄今为止，它们还不是艾滋病毒治疗方式的目标。一个 对病毒-宿主复合物的功能和机制的更好的分子理解可能会揭示新的 干预的治疗策略，包括宿主导向的治疗策略，这可能会逃避 当前药物治疗方案的局限性，其中靶向 HIV 酶的突变会降低药物疗效。 HARC 中心将使用集成的“系统到- Structure”平台，包括 (1) 通过功能蛋白质组学和遗传学的新方法进行发现 正在 HARC 中心开发，(2) 通过突破性 CRISPR 方法在原代 T 细胞中进行验证 以及有针对性的生化和功能测定，以及 (3) 使用合成物进行结构测定 HARC 中心之前的迭代中开发的创新结构技术，旨在解决大型、 我们研究灵活的、异质的、有时甚至是与膜相关的系统。 拟议的研究项目围绕三个服务于中心目标的主题。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）并通过三个项目研究病毒宿主进化（主题 3）：Vif 的结构和进化 和 APOBEC3（项目 1）、HIV 转录和潜伏期的调控（项目 2）以及遗传学和 进化（项目3）。 HARC中心项目由涵盖蛋白质组学的4个技术核心支持 方法（核心 1 - 蛋白质组学）、CRISPR 筛选和原代细胞内源标记来研究病毒- 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 HIV 病毒-宿主复合物结构的抗体技术（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、进展情况 行政以及外展活动和沟通将由行政核心监督 （核心 5）。发展核心（核心 6）将为年轻研究者和 HARC 提供培训机会 中心成员并授予合作机会基金，以增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.