HIV Gag Precursor Protein Interactions

 描述(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，并开发新的方法来抑制其复制。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蛋白是很有吸引力的靶标，因为它们在生命周期中扮演着多种角色。这些蛋白最初被合成为N-末端肉豆蔻酰化前体(PrGag)蛋白，利用其N-末端基质(MA)结构域靶向递送到质膜(PM)病毒组装部位。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、鞘磷脂和瑞瑞德。MA还与RNA结合，提出了一种模型，在该模型中，RNA结合保护MA在PrGag传递到PM富含PI(4，5)P2的位置之前不与不适当的细胞膜结合除了它的运输作用外，MA还被证明介导HIV-1包膜(Env)糖蛋白复合体进入病毒颗粒。MA-Env相互作用涉及Env跨膜(TM；gp41)部分的长细胞质尾巴(CT)，以及MA三聚体远端(辐条)和界面区(Hub)的残基；但MA如何将Env组装成病毒粒子的模型仍是一个推测问题。以我们以前的研究和初步结果为基础，我们建议剖析MA-膜/RNA和MA-Env结合的机制，并表征它们的抑制方法。我们的结果将有助于阐明HIV组装机制是如何运作的；并将导致针对GAG的抗病毒药物的开发，并了解它们是如何工作的。为实现这些目标，我们的目标如下：1.HIV-1基质-膜/RNA结合的特性及其抑制。2.阐明HIV-1基质与包膜蛋白的相互作用。

 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.