Retrovirus Restriction by Trima5alpha

Trima5alpha 的逆转录病毒限制

• 批准号: 6999737
• 负责人: Paul D. Bieniasz
• 金额: $ 43.94万
• 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 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6999737
• 关键词: Retroviridae; alleles; animal genetic material tag; capsid; cell component structure /function; chimeric proteins; gene mutation; genetic manipulation; genetic mapping; genetic polymorphism; human genetic material tag; human immunodeficiency virus 1; immunity; intermolecular interaction; laboratory mouse; laboratory rabbit; microorganism immunology; protein binding; protein localization; protein structure; protein structure function; transport proteins; virus genetics; virus replication

DESCRIPTION (provided by applicant): The study of endogenous cellular factors that inhibit infection by retroviruses is an emerging area of research. We were among the first to show that such factors in monkey cells can inhibit primate lentivirus infection and contribute in a major way to defining retrovirus species tropism. The recent identification of TRIM5alpha as one of this class of retrovirus resistance factors makes this an appropriate time to expand our efforts to characterize these factors. We have obtained preliminary data indicating that TRIM5alpha from humans and nonhuman primates restricts infection by a number of retroviruses, as widely divergent as MLV and HIV-1, and that HIV-1 CA:cyclophilin interaction can modulate restriction factor sensitivity. Other preliminary findings suggest that TRIM5alpha may not be the only capsid-specific retrovirus restriction factor in human cells. This proposal has three specific aims: Aim 1 is to determine the breadth and specificity of retrovirus inhibitory activity exhibited by a number of TRIM5alpha variants and to map determinants of specificity in CA and TRIM5alpha. This will involve the construction of chimeric and mutant human and nonhuman primate TRIM5alpha variants and the selection of HIV-1 mutants that are resistant to TRIM5alpha variants that inhibit wild-type HIV-1. As well as illuminating determinants of restriction specificity in TRIM5alpha and CA, this will provide critical control reagents for Aim 2, which is to determine the mechanism of restriction by TRIM5alpha. We will determine the subcellular localization of TRIM5alpha, attempt to demonstrate physical interaction between HIV-1 capsids and TRIM5alpha, map sequences responsible for TRIM5alpha subcellular localization, multimerization, and capsid binding and determine whether they are required for restriction. We will also determine whether proteins that bind to TRIM5alpha are required for restriction and test the hypothesis that TRIM5alpha-catalyzed ubiquitination is part of the restriction mechanism. Finally, in Aim 3 we will determine whether TRIM proteins that are closely related to TRIM5alpha exhibit retrovirus restriction activity. The successful completion of these studies could have practical implications both for AIDS animal model development, gene therapy vector development and, eventually, for the designing completely novel therapeutic antiretroviral strategies.

描述（由申请人提供）：抑制逆转录病毒感染的内源性细胞因子的研究是一个新兴的研究领域。我们是第一个证明猴细胞中的此类因子可以抑制灵长类慢病毒感染并在很大程度上确定逆转录病毒物种向性的人之一。最近将 TRIM5alpha 鉴定为此类逆转录病毒抗性因子之一，这使得现在正是扩大我们的努力来表征这些因子的适当时机。我们获得的初步数据表明，来自人类和非人灵长类动物的 TRIM5alpha 限制了许多逆转录病毒的感染，如 MLV 和 HIV-1，并且 HIV-1 CA：亲环蛋白相互作用可以调节限制因子敏感性。其他初步发现表明 TRIM5alpha 可能不是人类细胞中唯一的衣壳特异性逆转录病毒限制因子。该提案有三个具体目标： 目标 1 是确定许多 TRIM5alpha 变体表现出的逆转录病毒抑制活性的广度和特异性，并绘制 CA 和 TRIM5alpha 特异性的决定因素。这将涉及嵌合和突变的人类和非人灵长类TRIM5α变体的构建，以及对抑制野生型HIV-1的TRIM5α变体具有抗性的HIV-1突变体的选择。除了阐明 TRIM5alpha 和 CA 中限制特异性的决定因素外，这还将为目标 2 提供关键的对照试剂，即确定 TRIM5alpha 的限制机制。我们将确定 TRIM5alpha 的亚细胞定位，尝试证明 HIV-1 衣壳和 TRIM5alpha 之间的物理相互作用，绘制负责 TRIM5alpha 亚细胞定位、多聚化和衣壳结合的序列，并确定它们是否需要限制。我们还将确定限制是否需要与 TRIM5alpha 结合的蛋白质，并测试 TRIM5alpha 催化的泛素化是限制机制的一部分的假设。最后，在目标 3 中，我们将确定与 TRIM5alpha 密切相关的 TRIM 蛋白是否表现出逆转录病毒限制活性。这些研究的成功完成可能对艾滋病动物模型的开发、基因治疗载体的开发以及最终设计全新的抗逆转录病毒治疗策略产生实际影响。