Role of SNARE proteins in HIV-1 assembly and release

描述(申请人提供)：病毒的基因数量有限，但生命周期复杂。因此，它们进化到利用许多宿主因素来完成它们的复制。关于HIV-1的组装，不同的病毒蛋白同步移动到质膜上的特定位置，使病毒粒子能够形成形态。宿主SNARE(可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体)蛋白广泛参与囊泡介导转运的晚期阶段。这些蛋白质催化囊泡和靶区中相对的膜的对接和融合。陷阱不仅参与胞内和胞外的运输途径，而且还参与其他膜融合和裂变事件。其中包括胞质分裂期间的子细胞分离，这一现象与HIV-1的萌发过程有着惊人的相似之处。通过对圈套分类机制的广泛破坏，我们最近证明了这些蛋白通过影响GAG定位到质膜而在HIV-1组装中发挥作用。初步数据表明，SNARE蛋白可能调节GAG与质膜结合/运输所需的细胞运输途径。在这项研究中，我们建议定义圈套干扰影响HIV颗粒产生的确切机制。我们的目标是研究圈套破坏对细胞蛋白质和脂肪成分的影响，这些蛋白质和脂肪成分对HIV颗粒的产生至关重要。我们还将确定圈套破坏是否会影响细胞膜结构的完整性。最后，我们建议研究在这一过程中是否有特定的“v”(囊泡相关)或“t”(靶膜相关)SNARE蛋白优先参与。这不仅将定义新宿主因子在HIV生命CCLE中的作用，而且更好地了解病毒组装和释放途径将有助于打开识别新的抗病毒靶点的途径。

DESCRIPTION (provided by applicant): Viruses have a limited number of genes but a complex life cycle. They have hence evolved to utilize numerous host factors to complete their replication. With regard to HIV-1 assembly, a synchronized movement of different viral proteins to specific sites on the plasma membrane enables virion morphogenesis. The host SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins are widely involved in late stages of vesicular mediated transport. These proteins catalyze the docking and fusion of apposing membranes in the vesicle and target compartment. The SNAREs are involved not only in the endocytic and exocytic trafficking pathways but also in other membrane fusion and fission events. These include daughter cell separation during cytokinesis, a phenomenon that bears striking similarity to the process of HIV-1 budding. By generalized disruption of the SNARE sorting machinery, we recently demonstrated a role for these proteins in HIV-1 assembly by affecting Gag localization to the plasma membrane. Preliminary data suggests that SNARE proteins may regulate the cellular trafficking pathways required for Gag association/transport with the plasma membrane. In this study, we propose to define the precise mechanism via which SNARE disruption affects HIV particle production. We aim to study the effects of SNARE disruption on the cellular proteins and lipid components important for HIV particle production. We will also determine if SNARE disruption affects the integrity of the cellular membrane architecture. Finally, we propose to study whether there is a preferential involvement of specific "v" (vesicle-associated) or "t" (target membrane-associated) SNARE proteins in this process. This will not only define a role for novel host factors in the HIV life ccle but a better understanding of the virus assembly and release pathway will help open avenues for identifying novel anti-viral targets.