HIV Gag Precursor Protein Interactions

 描述（由申请人提供）：尽管在艾滋病诊断和治疗方面取得了很大进展，但艾滋病的持续流行需要继续努力了解艾滋病毒复制的各个方面，并开发新的抑制方法。为了实现这些目标，我们试图确定HIV-1结构蛋白（Gag）的活性和相互作用，以便设计干扰这些功能的抗病毒药物。Gag蛋白是有吸引力的靶标，因为它们在生命周期中发挥多种作用。蛋白质最初合成为N-末端豆蔻酰化前体（PrGag）蛋白，其利用其N-末端基质（MA）结构域靶向递送至质膜（PM）病毒组装位点。有证据表明，HIV-1 MA优先结合信号磷脂磷脂酰肌醇4，5二磷酸（PI[4，5]P2），并且病毒膜富含脂筏成分，如胆固醇、鞘磷脂和ceremide。MA还结合RNA，这表明了一种模型，其中RNA结合保护MA在PrGag递送至PM处的PI（4，5）P2富集位点之前不与不适当的细胞内膜结合。除了其运输作用外，MA还被证明介导HIV-1包膜（Env）糖蛋白复合物掺入病毒颗粒。MA-Env相互作用涉及Env的跨膜（TM; gp 41）部分的长胞质尾（CT），以及MA三聚体的远端（辐条）和界面区域（枢纽）处的残基;但是MA如何介导Env组装成病毒体的模型仍然是推测的问题。利用我们以前的研究和初步结果作为基础，我们建议解剖MA-膜/RNA和MA-Env结合的机制，并表征其抑制方法。我们的研究结果将有助于阐明HIV组装机制如何运作;并将导致开发以Gag为靶点的抗病毒药物，并了解它们如何工作。为了实现这些目标，我们的目标如下：1。HIV-1基质-膜/RNA结合及其抑制的表征。2. HIV-1基质-包膜蛋白相互作用的阐明。

 DESCRIPTION (provided by applicant): Despite great advances in AIDS diagnosis and treatment, the continuing AIDS epidemic demands continuing efforts to understand all aspects of HIV replication and to develop new methods for its inhibition. In pursuit of these goals, we have sought to define the activities and interactions of the HIV-1 structural (Gag) proteins so as to design antivirals that interfere with these functions. The Gag proteins are attractive targets since they perform multiple roles during the life cycle. The proteins initially are synthesized as N-terminally myristoylated precursor (PrGag) proteins that employ their N-terminal matrix (MA) domains to target delivery to plasma membrane (PM) virus assembly sites. Evidence indicates that the HIV-1 MA preferentially binds the signaling phospholipid phosphatidylinositol 4,5 bisphosphate (PI[4,5]P2), and that virus membranes are enriched for lipid raft constituents such as cholesterol, sphingomyelin, and ceremide. MA also binds RNA, suggesting a model in which RNA binding protects MA from binding to inappropriate intracellular membranes prior to PrGag delivery to PI(4,5)P2-rich sites at the PM. In addition to its trafficking role, MA also has been shown to mediate the incorporation of the HIV-1 envelope (Env) glycoprotein complex into virus particles. The MA-Env interaction involves the long cytoplasmic tail (CT) of the transmembrane (TM; gp41) portion of Env, and residues at the distal ends (spokes) and interface regions (hubs) of MA trimers; but models for how MA mediates Env assembly into virions remain a matter of speculation. Using our previous studies and preliminary results as a foundation, we propose to dissect the mechanisms of MA-membrane/RNA and MA-Env binding, and to characterize methods for their inhibition. Our results will help clarify how the HIV assembly machinery operates; and will lead to the development of Gag-targeted antivirals, and an understanding of how they work. To achieve these ends, our aims are as follows: 1. Characterization of HIV-1 matrix-membrane/RNA binding and its inhibition. 2. Elucidation of HIV-1 matrix-envelope protein interactions.