Nef Localization Complex Formation and Function

Nef 定位复合体的形成和功能

• 批准号: 6861106
• 负责人: DAVID M REKOSH
• 金额: $ 22.85万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6861106
• 关键词: HIV infections; chemical association; dimer; gene mutation; human immunodeficiency virus 1; membrane structure; protein structure; protein structure function; tissue /cell culture; virus cytopathogenic effect; virus protein

DESCRIPTION (provided by applicant): This application to the "Innovation Grants Program for AIDS Research" proposes studies to investigate mechanisms underlying the function of the HIV-1 Nef protein. Nef plays a critical role in HIV pathogenesis, but the precise reason for its importance is not understood. Understanding of the molecular basis by which Nef contributes to HIV pathogenesis could be crucial for the development for the development of new therapeutic interventions. There is accumulating evidence that the HIV-1 Nef protein self-associates into dimers and also possibly trimers. There is also some published evidence and new data presented in our Preliminary Results that suggest that such complex formation may be regulated by Nef's association with lipid raft membrane domains. At the present time, the biological significance of Nef self-association is not clear. However, many of Nefs cellular targets are themselves regulated by oligomerization and oligomerization is a common biological regulatory mechanism. Thus it is tempting to speculate that oligomerization plays a key role in Nef function. This proposal explores the biological importance of Nef-self association as well as the mechanisms that potentially regulate it. If the oligomeric state of Nef is shown to be important, the interface between adjacent Nef molecules could become a novel target for the design of anti-HIV drugs. The specific questions that this proposal will address are: Aim #1: Does the Nef Dimer Have Functional Significance "hisaim will address two important questions. 1) Do mutations between aa 105-123 that would be expected to disrupt the Nef dimer interface, actually disrupt Nef function and dimer formation? Can dimerization and Nef function of mutant Nef molecules, be restored with compensatory mutations? Aim #2: Is Formation of the Nef Dimer Regulated by the N-terminal region of Nef and Membrane/Raft Association? Studies will be performed both in vivo and in vitro to determine if the N-teminus of Nef interacts with the Nef core to regulate accessibility and if membrane association is involved in this process.

描述（由申请人提供）：这份“艾滋病研究创新资助计划”申请提出了研究 HIV-1 Nef 蛋白功能的潜在机制的研究。 Nef 在 HIV 发病机制中发挥着关键作用，但其重要性的确切原因尚不清楚。了解 Nef 导致 HIV 发病机制的分子基础对于开发新的治疗干预措施至关重要。 越来越多的证据表明，HIV-1 Nef 蛋白自我结合成二聚体，也可能形成三聚体。我们的初步结果中还提供了一些已发表的证据和新数据，表明这种复合物的形成可能受到 Nef 与脂筏膜域的关联的调节。目前，Nef 自关联的生物学意义尚不清楚。然而，许多 Nefs 细胞靶标本身受到寡聚化的调节，而寡聚化是一种常见的生物调节机制。因此，人们很容易推测寡聚化在 Nef 功能中发挥着关键作用。该提案探讨了 Nef-自关联的生物学重要性以及潜在的调节机制。如果 Nef 的寡聚状态被证明很重要，那么相邻 Nef 分子之间的界面可能成为抗 HIV 药物设计的新目标。该提案将解决的具体问题是： 目标#1：Nef 二聚体是否具有功能意义“hisaim 将解决两个重要问题。1) 预计会破坏 Nef 二聚体界面的 aa 105-123 之间的突变是否实际上会破坏 Nef 功能和二聚体形成？二聚化和突变的Nef分子的Nef功能，可以通过补偿性突变来恢复吗？ 目标#2：Nef 二聚体的形成是否受 Nef 的 N 末端区域和膜/筏关联调节？将在体内和体外进行研究，以确定 Nef 的 N 末端是否与 Nef 核心相互作用以调节可及性，以及膜关联是否参与该过程。