HIV-1 Modulation of the Ubiquitinome and Its Effects on Cell-to-Cell Transmission

HIV-1 泛素组的调节及其对细胞间传播的影响

• 批准号: 9187401
• 负责人: Mary Kathleen Lewinski
• 金额: $ 19.79万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9187401
• 关键词: Adverse effects; Affect; Anti-Retroviral Agents; Antiviral Agents; Biology of HIV Infection; CD4 Antigens; CD4 Positive T Lymphocytes; Cell Communication; Cell Line; Cell membrane; Cells; Cellular Membrane; Complement; Data; Data Set; Dependency; Development; Doxycycline; Engineering; Enhancers; Environment; Family; Fostering; Foundations; Funding Agency; Gene Expression; Genes; Giant Cells; Global Change; Goals; HIV; HIV therapy; HIV-1; Human; Immune response; Immunoprecipitation; Infection; Innate Immune Response; Interferons; Mass Spectrum Analysis; Membrane Proteins; Methods; Molecular Biology; Pathway interactions; Physicians; Proteins; Proteome; Proteomics; Proviruses; Recruitment Activity; Reporting; Research; Role; Scientist; Site; Surface; System; Testing; Tetanus Helper Peptide; Tetracyclines; Training; Trans-Activators; Ubiquitin; Ubiquitination; Viral; Viral Genes; Viral Proteins; Virion; Virus; Virus Replication; antiretroviral therapy; career; career development; cellular targeting; cytotoxic; multicatalytic endopeptidase complex; mutant; novel; promoter; protein degradation; public health relevance; suicide inhibitor; superinfection; trafficking; transmission process; ubiquitin ligase; vpu Genes; vpu Protein

 DESCRIPTION (provided by applicant): A broadly applicable cure for HIV-1 infection has remained elusive, in part, because the virus is able to circumvent host adaptive and innate immune responses. HIV-1 commandeers cellular pathways for its replication and antagonizes host proteins that inhibit viral replication. The HIV-1 accessory proteins, Vif, Vpr, Vpu and Nef, direct the degradation or mislocalization of cellular proteins that interfere with viral replicatio and persistence. Vif, Vpr and Vpu all usurp cellular ubiquitin ligases to degrade specific cellular proteins and facilitate replication. Some targets of these accessory proteins are known, but others are as yet undefined. Elucidating these viral-host cell interactions may reveal new viral dependencies on cellular proteins as well as new host antiviral proteins and viral countermeasures, revealing potential targets for antiretroviral therapy. The proposed project specifically examines the interaction between Vpu and the cellular ubiquitin pathway, determining the extent to which Vpu acts as a suicide inhibitor when targeting cellular proteins such as CD4 for degradation, the consequences these effects have for HIV-1 replication including viral cell-to-cell transmission. In addition, this study develops a novel experimental system to investigate the effects of HIV-1 gene expression and Vpu specifically on the modulation of cellular membrane proteins and global ubiquitination and degradation of cellular proteins, with the goal of attributing particular effects to the accessory gene vpu in the context f the complete repertoire of viral gene expression. In determining the host cell proteins modulated by HIV-1, this study will provide a better understanding of how HIV-1 manipulates the cellular environment to facilitate its replication and evade the immune response, which in turn will inform the development of new antiretroviral therapies. This proposal is intended to support the career development and further training of the candidate, who aims to extend her expertise in the molecular biology of HIV infection to include proficiency in the acquisition and analysis of proteomics and ubiquitinomics data sets. This project is expected to identify new cellular targets of HIV that will facilitate the development of independent lines of research, providing the foundation for the candidate to pursue additional sources of funding and foster a career as an independent physician- scientist.

 描述(由申请人提供)：HIV-1感染的广泛适用的治疗方法仍然难以捉摸，部分原因是该病毒能够绕过宿主适应性和先天免疫反应。HIV-1占据了其复制的细胞途径，并对抗抑制病毒复制的宿主蛋白。HIV-1的辅助蛋白Vif、Vpr、VPu和Nef指导细胞蛋白的降解或错误定位，从而干扰病毒的复制和持续。VIF、VPR和VPU都篡夺细胞泛素连接酶来降解特定的细胞 蛋白质和促进复制。这些辅助蛋白的一些靶点是已知的，但其他的还不确定。阐明这些病毒-宿主细胞相互作用可能揭示新的病毒对细胞蛋白的依赖以及新的宿主抗病毒蛋白和病毒对策，揭示抗逆转录病毒治疗的潜在靶点。这项拟议的项目具体研究了VPU和细胞泛素途径之间的相互作用，确定了VPU在针对细胞蛋白质(如CD4)进行降解时充当自杀抑制因子的程度，以及这些影响对HIV-1复制(包括病毒细胞间传播)的影响。此外，本研究还开发了一种新的实验系统来研究HIV-1基因的表达和VPU对细胞膜蛋白的调节以及细胞蛋白的全球泛素化和降解的影响，目的是在完整的病毒基因表达谱的背景下将特定的影响归因于辅助基因VPU。在确定HIV-1调节的宿主细胞蛋白时，这项研究将提供更好的理解，了解HIV-1如何操纵细胞环境以促进其复制和逃避免疫反应，这反过来将为新的抗逆转录病毒疗法的开发提供信息。这项提议旨在支持候选人的职业发展和进一步培训，她的目标是扩大她在艾滋病毒感染的分子生物学方面的专业知识，包括熟练地获取和分析 蛋白质组学和泛素组学数据集。该项目预计将确定艾滋病毒的新细胞靶点，以促进独立研究领域的发展，为候选人寻求更多资金来源和培养独立内科科学家的职业生涯奠定基础。