Discovery and Mechanism of Antiretroviral Factors

抗逆转录病毒因子的发现及其机制

• 批准号: 8415561
• 负责人: Paul D. Bieniasz
• 金额: $ 43.71万
• 依托单位: AARON DIAMOND AIDS RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-01 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8415561
• 关键词: Acquired Immunodeficiency Syndrome; Animal Model; Animals; Anti-Retroviral Agents; Antiretroviral resistance; Antiviral Agents; Base Sequence; Biological Assay; Candidate Disease Gene; Capsid; Cells; Communicable Diseases; Data; Defense Mechanisms; Engineering; Event; Exhibits; Funding; Fv-1 protein; Gene Library; Genes; Goals; Grant; HIV; HIV-1; Host Defense; Human; Infection; Interferon Type I; Intervention; Laboratory Animals; Lead; Life Cycle Stages; Macaca; Macaca mulatta; Modeling; Nucleic acid sequencing; Organism; Pharmacologic Substance; Physiological; Property; Proteins; Relative (related person); Resistance; Retroviridae; SIV; Sequence Analysis; Staging; Surface; Techniques; Testing; Time; Variant; Viral; Virus; Work; animal model development; base; cDNA Library; cell type; gene discovery; high reward; high risk; inhibitor/antagonist; mutant; particle; prevent; public health relevance; selective expression; tool

DESCRIPTION (provided by applicant): The ancestors of modern organisms were colonized by viruses that were very rarely beneficial, and often deleterious to the host. Inevitably, hosts adapted to limit viral replication, and in so doing evolved various autonomous antiviral defense mechanisms. One of these is TRIM5, which is multimeric protein that recognizes incoming retroviral capsids, but its precise mechanisms of action are unclear. In specific aim 1 we will develop new techniques to understand what happens to various incoming components of retroviral particles as they complete the early steps of their life cycle. These assays will very likely illuminate the mechanisms by which TRIM5 proteins and other antiretroviral factors exert their inhibitory effects. Moreover, these techniques are likely to have a wide range of applications in the study of these steps of the retroviral life cycle. In specific aim 2 we will derive TRIM5-resistant HIV-1 capsids that will identify residues on the surface of the HIV-1 capsid that are targeted by TRIM5 and will serve a useful tools to probe the mechanism of action of TRIM5. Additionally these capsids will be used for generating simian-tropic HIV- strains that can replicate in rhesus macaques. In addition to known antiretroviral proteins like TRIM5, there are several reasons to think that there are more, perhaps many more, antiviral host defense genes in to be discovered. In specific aim 3, we will attempt to identify such genes by constructing focused, arrayed cDNA libraries, each consisting of tens to hundreds of candidate genes, that either (i) have key properties exhibited by known antiretroviral genes or (ii) are selectively expressed in cells that exhibit constitutive or induced resistance to HIV-1 or SIV infection. These genes libraries will be arrayed each gene will be individually tested for its ability to inhibit retrovirus replication.

描述（由申请人提供）：现代生物的祖先被很少有益且经常对宿主有害的病毒殖民。不可避免地，适合限制病毒复制的宿主，这样做就进化出了各种自主抗病毒防御机制。其中之一是TRIM5，它是多聚体蛋白，识别传入的逆转录病毒衣壳，但其精确的作用机理尚不清楚。在特定的目标1中，我们将开发新技术，以了解逆转录病毒颗粒的各种传入组成部分在完成生命周期的早期步骤时会发生什么。这些测定很可能会阐明TRIM5蛋白和其他抗逆转录病毒因子的抑制作用的机制。此外，这些技术在研究逆转录病毒生命周期的这些步骤中可能具有广泛的应用。在特定的目标2中，我们将得出抗TRIM5的HIV-1衣壳，这些CAPSID将识别由TRIM5靶向的HIV-1 CAPSID表面上的残基，并将提供有用的工具来探测TRIM5的作用机理。另外，这些衣壳将用于生成可以在恒河猕猴中复制的邻苯二甲酸菌株。除了已知的抗逆转录病毒蛋白（如TRIM5）外，还有几个理由认为，还可以发现更多（也许还有更多的抗病毒宿主防御基因）。在特定的目标3中，我们将尝试通过构建聚焦的，有阵列的cDNA库来识别此类基因，每个文库由数十个至数百个候选基因组成，即（i）具有已知抗逆转录病毒基因或（ii）的关键特性，或（ii）在细胞中选择性表达，表现出对HIV抗HIV或诱导HIV-1或SIV感染的抗性。这些基因文库将被批准，每个基因都将分别测试其抑制逆转录病毒复制的能力。