The Role of Nef and Cyclophilin A in HIV-1 Disassembly

Nef 和亲环蛋白 A 在 HIV-1 分解中的作用

• 批准号: 6896429
• 负责人: MICHAEL D POWELL
• 金额: $ 17.75万
• 依托单位: MOREHOUSE SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-01 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6896429
• 关键词: chimeric proteins; enzyme linked immunosorbent assay; human immunodeficiency virus 1; human immunodeficiency virus 2; mass spectrometry; nucleocapsid; peptidylprolyl isomerase; polymerase chain reaction; protein binding; protein protein interaction; simian immunodeficiency virus; virion; virus assembly; virus infection mechanism; virus protein; virus replication; western blottings

DESCRIPTION (provided by applicant): The development of new HIV-1 antivirals is dependent on our understanding of the individual steps in viral replication. Perhaps the least understood aspect of HIV-1 replication is the process of disassembly. Our preliminary data suggest that the viral protein Nef and the cellular factor cyclophilin A (CyPA) are involved in some aspect of disassembly. We have shown that HIV-1 viruses that have their own nef deleted and replaced by HIV-2 or SIV Nef "in trans" become resistant to cyclosporine A treatment. This suggests some interdependence between Nef and CyPA. It has been previously shown that Nef and CyPA can directly interact. We have confirmed this result in our own lab using Far Western blots and SELDI mass spectrometry. We consider whether the interactions between Nef and CyPA and their potential interactions with CA in the viral core are important for replication. Our hypothesis is: that interaction between Nef, CyPA and CA are important for enhancement of viral infectivity. Our approach is summarized in three specific aims: 1. Characterize the ability of HIV-2 and SIV Nef to induce cyclophilin A independence in HIV-1 virions using a multi-round infectivity assay. 2. Determine if the interaction between HIV-1 Nef and CyPA is relevant to enhancement of infectivity. 3. Determine if the HIV-1, HIV-2 or SIV Nef proteins can directly or indirectly bind to CA protein. A better understanding of the relationship between Nef and CyPA will provide critical details about this poorly characterized step in replication, which could provide an attractive new target for antiviral therapy.

描述（由申请人提供）：新型 HIV-1 抗病毒药物的开发取决于我们对病毒复制各个步骤的理解。也许 HIV-1 复制最不为人所知的方面是分解过程。我们的初步数据表明，病毒蛋白 Nef 和细胞因子亲环蛋白 A (CyPA) 参与了分解的某些方面。我们已经证明，删除自身 nef 并“反式”被 HIV-2 或 SIV Nef 取代的 HIV-1 病毒会对环孢菌素 A 治疗产生耐药性。这表明 Nef 和 CyPA 之间存在一定的相互依赖性。之前已经证明 Nef 和 CyPA 可以直接相互作用。我们已经在我们自己的实验室中使用 Far Western 印迹和 SELDI 质谱证实了这一结果。我们考虑 Nef 和 CyPA 之间的相互作用以及它们与病毒核心中 CA 的潜在相互作用是否对复制很重要。我们的假设是：Nef、CyPA 和 CA 之间的相互作用对于增强病毒感染性很重要。我们的方法概括为三个具体目标： 1. 使用多轮感染性测定来表征 HIV-2 和 SIV Nef 在 HIV-1 病毒粒子中诱导亲环蛋白 A 独立性的能力。 2. 确定 HIV-1 Nef 和 CyPA 之间的相互作用是否与感染性增强相关。 3.确定HIV-1、HIV-2或SIV Nef蛋白是否可以直接或间接结合CA蛋白。更好地理解 Nef 和 CyPA 之间的关系将提供有关复制过程中这一特征不明的步骤的关键细节，这可能为抗病毒治疗提供一个有吸引力的新靶点。