HARC Center: HIV Accessory and Regulatory Complexes

HARC 中心：HIV 辅助和调节复合物

• 批准号: 8726997
• 负责人: ALAN D FRANKEL
• 金额: $ 410.06万
• 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 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8726997
• 关键词: Acquired Immunodeficiency Syndrome; Antiviral Agents; Biological; Cell Cycle; Cells; Complex; Core-Binding Factor; Coupled; Degradation Pathway; Drug resistance; Enzymes; Funding Opportunities; Genetic Transcription; Goals; HIV; Host Defense; Individual; Infection; Integration Host Factors; Life Cycle Stages; Life Expectancy; Maps; Mass Spectrum Analysis; Measures; Mediating; Membrane Proteins; Methods; Modeling; Molecular; N-terminal; Nuclear Export; Nucleic Acids; Peptide Hydrolases; Positive Transcriptional Elongation Factor B; Post-Translational Protein Processing; Proteinase 3; Proteins; Proteomics; RNA; Recruitment Activity; Regulation; Role; Small Nuclear Ribonucleoproteins; Specificity; Structure; System; Technology; Therapeutic; Therapeutic Intervention; Ubiquitination; Viral; Virus; Virus Diseases; base; cofactor; genetic regulatory protein; innovation; novel; protein complex; protein degradation; trafficking; ubiquitin-protein ligase; vif Gene Products; viral RNA; virology

DESCRIPTION (provided by applicant): HIV requires the host cell machinery for replication. Many complexes hijacked by HIV have been identified, but structures are known in only a few cases. The HARC Center is taking a broad systems-to-structure approach to this problem, having identified and validated new host complexes through a comprehensive proteomics effort. Primary biological aims of the Center are to achieve comprehensive structural pictures of: (1) how the accessory proteins Vif, Vpu, Vpr, and the viral protease PR disarm host defenses and circumvent viral restriction via degradation pathways, and (2) how the regulatory proteins Tat and Rev hijack the host transcription and RNA trafficking machinery to express and package viral RNAs. There is growing evidence that HIV accessory proteins primarily target host antiviral restriction factors for destruction. For Vif, we will determine the structures of the E3 ligase complex and interactions with APOBEC, and evaluate the functional roles of the CBF? cofactor and post-translational modifications (PTMs). For Vpu, we will determine the structures of restriction factor complexes and map effects of Vpu on ubiquitination. For Vpr, we will validate new host interactions and mechanisms and assemble complexes for structure determination. For PR, we will determine the structures and functions of new host target complexes and measure the levels of PR activity during infection. The regulatory proteins Tat and Rev hijack host transcription and RNA export machineries. For Tat, we will determine the structures of newly discovered AFF4 elongation complexes, and characterize other host factors, PTMs, and inhibitory 7SK snRNP complexes. For Rev, we will determines the structures of Rev-RRE nuclear export complexes, map viral RNA structures, and characterize the roles of new host proteins in post export functions. The HARC Center also relies on technology innovation from four cores. The EM Core will develop methods to determine structures of membrane protein complexes, and use Fabs to solve the structures of small HIV-host complexes. The Proteomics Core will extend mass spectrometry analyses to host protein complexes and map HIV-dependent host PTMs. The Computational Core will develop methods to characterize allostery and detailed models to study PTMs. The Virology Core will measure effects of new host interactions on HIV replication and coupled activities of Tat and Rev. The individual projects and technologies depend critically on an extensive network of collaborators, which will be expanded through a Collaborative Opportunity Fund.

描述（由申请人提供）：HIV 需要宿主细胞机制进行复制。许多被艾滋病毒劫持的复合物已被识别，但只有少数情况下的结构为人所知。 HARC 中心正在采取广泛的系统到结构的方法来解决这个问题，通过全面的蛋白质组学工作鉴定并验证了新的宿主复合物。该中心的主要生物学目标是获得以下方面的全面结构图：(1) 辅助蛋白 Vif、Vpu、Vpr 和病毒蛋白酶 PR 如何通过降解途径解除宿主防御并规避病毒限制，以及 (2) 监管如何Tat 和 Rev 蛋白劫持宿主转录和 RNA 运输机制来表达和包装病毒 RNA。越来越多的证据表明，HIV 辅助蛋白主要针对宿主抗病毒限制因子进行破坏。对于 Vif，我们将确定 E3 连接酶复合物的结构以及与 APOBEC 的相互作用，并评估 CBF 的功能作用？辅因子和翻译后修饰 (PTM)。对于Vpu，我们将确定限制因子复合物的结构并绘制Vpu 对泛素化的影响。对于 Vpr，我们将验证新的宿主相互作用和机制，并组装复合物以进行结构确定。对于 PR，我们将确定新宿主靶复合物的结构和功能，并测量感染期间 PR 活性的水平。调节蛋白 Tat 和 Rev 劫持宿主转录和 RNA 输出机器。对于 Tat，我们将确定新发现的 AFF4 延伸复合物的结构，并表征其他宿主因子、PTM 和抑制性 7SK snRNP 复合物。对于 Rev，我们将确定 Rev-RRE 核输出复合物的结构，绘制病毒 RNA 结构图，并表征新宿主蛋白在输出后功能中的作用。 HARC中心还依赖于四个核心的技术创新。 EM Core 将开发确定膜蛋白复合物结构的方法，并使用 Fab 来解析小型 HIV 宿主复合物的结构。蛋白质组学核心将质谱分析扩展到宿主蛋白质复合物并绘制 HIV 依赖性宿主 PTM 图谱。计算核心将开发表征变构的方法和研究 PTM 的详细模型。病毒学核心将测量新宿主相互作用对 HIV 复制以及 Tat 和 Rev 联合活动的影响。各个项目和技术在很大程度上依赖于广泛的合作者网络，该网络将通过合作机会基金进行扩展。