Elucidation of HIV Env acquisition strategies

描述(由申请人提供)：创建具有感染性的HIV-1颗粒的一个必要步骤是掺入表面融合糖蛋白(对于逆转录病毒，称为Env)。只有当Env运输到作为病毒组装部位的细胞膜的精确斑块时，这种掺入才会发生。促进病毒糖蛋白转位的病毒组装部位的特征仍然是一个谜。我们已经证明，来自不同病毒家族的糖蛋白可以有效地招募到人类免疫缺陷病毒(HIV-1)组装部位，但大多数宿主蛋白不是。我们假设，被包裹的病毒利用一种共同的机制来促进病毒糖蛋白的重新分布到病毒组装部位。由于许多募集到HIV-1组装位点的病毒糖蛋白与天然的HIV-1糖蛋白(HIV-1 env)没有序列相似性，因此不太可能通过HIV之间的直接蛋白质-蛋白质相互作用来促进外来糖蛋白募集 1结构蛋白(GAG)和糖蛋白。因此，这项建议的中心目标是(1)确定病毒糖蛋白中决定其吸引病毒组装部位的特征(S)，以及(2)鉴定有助于其吸引的病毒组装部位的特征(S)。这项提议包含四个具体目标。目的1：确定不同的病毒糖蛋白是否共包装成相同的病毒颗粒。目的2：利用逆转录病毒突变文库来确定病毒糖蛋白的哪些特征决定了它们对病毒组装部位的吸引力。目的3：实时进行糖蛋白采集的定量成像。目的4：应用正选择方法确定GAG与糖蛋白CTDS的配伍决定因素。这些目标将有助于确定调节HIV-1生命周期中这一关键步骤的相互作用。

DESCRIPTION (provided by applicant): A requisite step for the creation of an infectious HIV-1 particle is incorporation of the surface fusion glycoprotein (for retroviruses, referred to as Env) This incorporation only occurs if Env traffics to the precise patch of membrane in the cell that serves as the viral assembly site. The features of the viral assembly site that facilitate this translocation of viral glycoproteins remain a mystery. We have demonstrated that glycoproteins from diverse families of viruses are efficiently recruited to human immunodeficiency virus (HIV-1) assembly sites but that most host proteins are not. We hypothesize that enveloped viruses utilize a common mechanism to facilitate the redistribution of viral glycoproteins to viral assembly sites. Because many of the viral glycoproteins recruited to HIV-1 assembly sites contain no sequence similarity with the native HIV-1 glycoprotein (HIV-1 Env), it is unlikely that foreign glycoprotein recruitment is facilitated by a direct protein-protein interaction between HIV 1 structural protein (Gag) and the glycoprotein. The central objectives of this proposal, therefore, are (1) to identify the feature(s) in viral glycoproteins that dictate their attraction o viral assembly sites and (2) to identify the feature(s) of the viral assembly site that facilitate his attraction. This proposal contains four specific aims. Aim 1: Determine whether diverse viral glycoproteins co-package into the same viral particles. Aim 2: Utilize retroviral mutagenesis libraries to ascertain what features in viral glycoproteins dictate their attraction to viral assemly sites. Aim 3: Perform quantitative imaging of glycoprotein acquisition in real time. Aim 4: Apply positive selection to identify determinants of compatibility between Gag and glycoprotein CTDs. These aims will help define the interactions that modulate this critical step in the HIV-1 lifecycl.