Mechanisms that determine subcellular sites of HIV-1 assembly

决定 HIV-1 组装亚细胞位点的机制

• 批准号: 9423742
• 负责人: Akira Ono
• 金额: $ 56.12万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9423742
• 关键词: Acidic Region; Acquired Immunodeficiency Syndrome; Address; Antiviral Agents; Binding; CD44 gene; Cell membrane; Cells; Cytoplasm; Data; Distant; Ensure; Factor V; Failure; Goals; HIV-1; In Vitro; Infection; Integration Host Factors; Knowledge; Life Cycle Stages; Lipid Binding; Lipids; Location; Mediating; Membrane; Membrane Proteins; Microscopy; Modeling; Molecular; Movement; N-terminal; Outcome Study; Phase; Phospholipids; Process; Production; Property; Recruitment Activity; Research; Resolution; Role; Signal Transduction; Site; Staging; Stromal Cells; Structural Protein; T-Lymphocyte; Testing; Transfer RNA; Translation Process; Translations; Viral; Viral Structural Proteins; Virion; Virus; Virus Assembly; base; cell type; design; env Glycoproteins; evidence base; experimental study; extracellular; gag Gene Products; in vitro Assay; lymph nodes; novel; particle; viral transmission

Title: Mechanisms that determine subcellular sites of HIV-1 assembly Summary/Abstract: Virus particle assembly of HIV-1, the causative agent of AIDS, takes place at the plasma membrane (PM) in most cell types including natural host T cells. This process is driven by a viral structural protein Gag. The N- terminal matrix (MA) domain of Gag determines Gag localization to and hence virus assembly at the PM. MA mediates membrane binding of Gag via N-terminal myristoyl moiety and a highly basic region (HBR) that binds acidic lipids. Binding of HBR to a PM-specific acidic phospholipid PI(4,5)P2 is critical for PM localization of Gag and efficient virus release. Notably, in vitro and cell-based studies showed that MA HBR also interacts with tRNAs, which suppress binding of Gag to non-PI(4,5)P2 acidic lipids, suggesting tRNAs as a new host factor that regulates MA-membrane interactions. However, molecular determinants for the tRNA-MA HBR interaction and its reversal by the interaction with PI(4,5)P2, combination of which regulates PM-specific Gag localization, remain to be elucidated. Moreover, how these interactions begin and what effect these interactions have on the property of progeny virions at the end are poorly understood. Binding of tRNAs to MA HBR is most likely to occur at translation sites due to limited availability of tRNAs outside of the translation machinery. However, little is known about subcellular sites of Gag translation, where Gag begins its movement to the PM. At the PM, Gag multimerization is likely to create accumulation of acidic lipids, but its impact on the ability of progeny virions to spread to uninfected cells remains unknown. Our long-term goal is to elucidate mechanisms that determine sites of HIV-1 assembly and to use the knowledge for developing antiviral strategies. Our central hypothesis in this application is that MA HBR interactions with tRNAs, which begin during translation, and with acidic lipids determine subcellular Gag localization and the properties of progeny virions. To test this hypothesis, we plan to: 1) identify molecular determinants for interaction of MA HBR with tRNAs and its reversal by PI(4,5)P2; 2) identify tRNAs that suppress PI(4,5)P2-independent membrane binding but allow PI(4,5)P2-mediated reversal; 3) examine the possibility that Gag associates with tRNA during translation; and 4) examine the effect of acidic lipid accumulation at virus assembly sites on the virion properties. The knowledge gained from experiments outlined in this proposal will likely help us develop antiviral strategies that target mechanisms regulating Gag localization to the PM, thereby inhibiting extracellular virus release and spread.

标题：确定HIV-1组件亚细胞位点的机制 摘要/摘要： HIV-1的病毒颗粒组装是AIDS的致病药物，发生在质膜（PM）的 大多数细胞类型在内，包括天然宿主T细胞。该过程由病毒结构蛋白插入驱动。然后- GAG的末端基质（MA）结构域确定了插科打到PM处的病毒组装。马 通过N末端肉豆蔻酰基部分和高度碱性区域（HBR）介导GAG的膜结合 酸性脂质。 HBR与PM特异性酸性磷脂Pi（4,5）P2的结合对于PM定位至关重要 和有效的病毒释放。值得注意的是，基于细胞和细胞的研究表明，MA HBR也与 TRNA，抑制GAG与非PI（4,5）P2酸性脂质的结合，表明TRNA作为新的宿主因子 调节MA膜相互作用。但是，tRNA-MA HBR相互作用的分子决定因素 以及与PI（4,5）P2的相互作用的逆转，其组合调节PM特异性插入定位， 仍然有待阐明。此外，这些相互作用是如何开始的，这些相互作用对 最终，后代病毒体的特性知之甚少。 TRNA与MA HBR的结合最有可能 由于TRNA的可用性有限，因此在翻译机构外部的可用性有限。但是，很少 关于GAG翻译的亚细胞位点已知，其中插科打术开始向PM运动。在下午，插科打 多聚化可能会产生酸性脂质的积累，但其对后代病毒体能力的影响 扩散到未感染的细胞仍然未知。 我们的长期目标是阐明确定HIV-1组装位点的机制，并使用 开发抗病毒策略的知识。我们在此应用中的中心假设是Ma HBR 与TRNA的相互作用，该TRNA在翻译期间开始，与酸性脂质确定了亚细胞GAG 后代病毒体的定位和特性。为了检验该假设，我们计划：1）确定分子 MA HBR与TRNA相互作用的决定因素及其逆转PI（4,5）P2的决定因素； 2）确定trnas 抑制PI（4,5）P2非依赖性膜结合，但允许Pi（4,5）P2介导的逆转； 3）检查 翻译过程中GAG与tRNA相关的可能性； 4）检查酸性脂质的影响 病毒组装位点积聚在病毒素特性上。从概述的实验中获得的知识 在此提案中，可能会帮助我们制定针对调节堵嘴机制的抗病毒策略 将其定位到PM，从而抑制细胞外病毒释放和扩散。