Elucidation of HIV Env acquisition strategies

描述（由申请人提供）：产生感染性HIV-1颗粒的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运输到细胞中作为病毒组装位点的精确膜斑时，才会发生这种掺入。促进病毒糖蛋白易位的病毒组装位点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被有效地募集到人类免疫缺陷病毒（HIV-1）的组装位点，但大多数宿主蛋白不是。我们假设包膜病毒利用一种共同的机制来促进病毒糖蛋白重新分布到病毒组装位点。由于许多募集到HIV-1装配位点的病毒糖蛋白与天然HIV-1糖蛋白（HIV-1 Env）不含序列相似性，因此HIV-1和HIV-1之间的直接蛋白质-蛋白质相互作用不太可能促进外源糖蛋白的募集。 1结构蛋白（Gag）和糖蛋白。因此，该建议的中心目标是（1）鉴定病毒糖蛋白中决定其吸引病毒组装位点的特征，以及（2）鉴定病毒组装位点中促进其吸引的特征。这项建议包括四个具体目标。目的1：确定不同的病毒糖蛋白是否共同包装成相同的病毒颗粒。目的2：利用逆转录病毒突变库来确定病毒糖蛋白的哪些特征决定了它们对病毒组装位点的吸引力。目的3：进行糖蛋白采集的真实的实时定量成像。目的4：应用阳性选择来鉴定Gag和糖蛋白CTD之间相容性的决定因素。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.