HARC Center: HIV Accessory and Regulatory Complexes

HARC 中心：HIV 辅助和调节复合物

• 批准号: 8410292
• 负责人: ALAN D FRANKEL
• 金额: $ 415.4万
• 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/8410292
• 关键词: HARC; Center; HIV; Accessory; Regulatory

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. PUBLIC HEALTH RELEVANCE: Existing anti-HIV therapeutics have extended the life expectancy of infected individuals, however major limitations remain, including drug resistance. By determining structures of key HIV accessory and regulatory complexes, we will achieve a more complete molecular understanding of how the virus hijacks the host cell machinery, and also identify new targets for therapeutic intervention in the continuing battle against AIDS.

描述（由申请人提供）：艾滋病毒需要主机细胞机械进行复制。已经确定了许多被HIV劫持的复合物，但仅在少数情况下才知道结构。 HARC中心通过全面的蛋白质组学努力确定并验证了新的宿主络合物，正在采取广泛的系统对结构方法。 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.越来越多的证据表明，艾滋病毒附件蛋白主要针对宿主抗病毒药限制因素造成破坏。对于VIF，我们将确定E3连接酶复合物的结构和与APOBEC的相互作用，并评估CBF的功能作用？辅因子和翻译后修饰（PTMS）。对于VPU，我们将确定限制因子复合物的结构以及VPU对泛素化的效果。对于VPR，我们将验证新的主机相互作用和机制，并组装结构确定的复合物。对于PR，我们将确定新宿主目标复合物的结构和功能，并测量感染过程中PR活性的水平。调节蛋白TAT和Rev Hijack主机转录和RNA导出机械。对于TAT，我们将确定新发现的AFF4伸长复合物的结构，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，MAP病毒RNA结构的结构，并表征新的宿主蛋白在POST出口功能中的作用。 HARC中心还依靠四个核心的技术创新。 EM核心将开发确定膜蛋白复合物结构的方法，并使用FAB来求解小型HIV宿主复合物的结构。蛋白质组学核心将将质谱分析扩展到宿主蛋白质复合物，并绘制依赖HIV依赖性的宿主PTM。计算核心将开发出表征变构和详细模型来研究PTM的方法。病毒学核心将衡量新宿主相互作用对TAT和REV的艾滋病毒复制和耦合活动的影响。各个项目和技术急剧依赖于广泛的合作者网络，该网络将通过协作机会基金进行扩展。 公共卫生相关性：现有的抗HIV治疗方法扩大了感染者的预期寿命，但是仍然存在重大局限性，包括耐药性。通过确定关键的HIV辅助和调节络合物的结构，我们将对病毒如何劫持宿主细胞机械的方式进行更完整的分子理解，并确定在持续与艾滋病战斗中进行治疗干预的新目标。