Mechanisms of cell motility inhibition by the HIV-1 pathogenesis factor NEF

NEF蛋白是人类免疫缺陷病毒（HIV）的一种病毒编码致病因子，可促进病毒复制和受感染宿主中产生病毒的细胞的免疫逃避。我们之前确定了NEF对宿主细胞运动的抑制是一种高度保守的生物学特性，并确定了对肌动蛋白重塑的抑制是一种必要但不充分的分子机制。本建议的目的是定义NEF用于调节细胞运动的与肌动蛋白无关的活动。这将通过结合表达NEF的斑马鱼原始生殖细胞（PGC）的体内分析和病毒感染的原代人T淋巴细胞和巨噬细胞的体外研究来实现。先进的实时成像方法将在单个细胞水平上提供NEF诱导的缺陷的详细特征，并将直接与细胞运动的量化相结合。伴随的遗传和生化分析旨在揭示潜在的分子机制。这些跨学科的研究将利用NEF作为一个有价值的工具来解读PGC迁移的基本原理，并有望对宿主细胞运动的机制和病理生理影响产生重要的见解。

The NEF protein is a virally encoded pathogenesis factor of Human Immunodeficiency Viruses (HIV) that promotes virus replication and immune evasion of virus producing cells in the infected host. We previously identified inhibition of host cell motility as a highly conserved biological property of NEF and established inhibition of actin remodeling as a molecular mechanism that is necessary but not sufficient for this activity. The aim of this proposal is to define the actin-independent activities NEF employs to modulate cell motility. This will be achieved by combining in vivo analyses of NEF- expressing zebrafish primordial germ cells (PGC) with ex vivo studies of virally infected primary human T lymphocytes and macrophages. Advanced real time imaging approaches will provide detailed characterization of the defects induced by NEF on a single cell level and will be directly coupled to quantification of cell motility. Concomitant genetic and biochemical analyses are designed to unravel the underlying molecular mechanisms. Together these interdisciplinary studies will employ NEF as a valuable tool to decipher basic principles of PGC migration and are expected to yield important insights into the mechanism and patho-physiological impact on host cell motility.