Elucidation of HIV Env acquisition strategies

描述（由申请人提供）：创建感染性HIV-1颗粒的必要步骤是将表面融合糖蛋白（对于逆转录病毒，称为Env）结合，只有当Env运输到细胞中作为病毒组装位点的精确膜斑块时，这种结合才会发生。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可以有效地招募到人类免疫缺陷病毒（HIV-1）组装位点，但大多数宿主蛋白却不能。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配到病毒组装位点。由于招募到HIV-1组装位点的许多病毒糖蛋白与天然HIV-1糖蛋白（HIV-1 Env）没有序列相似性，因此外来糖蛋白的招募不太可能是由HIV之间的直接蛋白-蛋白相互作用促进的

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.