Department of Medicine, University of Cambridge, Cambridge, United Kingdom.
Cambridge Institute for Medical Research, University of Cambridge, Cambridge, United Kingdom.
Elife. 2019 Mar 12;8:e41431. doi: 10.7554/eLife.41431.
Viruses manipulate host cells to enhance their replication, and the identification of cellular factors targeted by viruses has led to key insights into both viral pathogenesis and cell biology. In this study, we develop an HIV reporter virus (HIV-AFMACS) displaying a streptavidin-binding affinity tag at the surface of infected cells, allowing facile one-step selection with streptavidin-conjugated magnetic beads. We use this system to obtain pure populations of HIV-infected primary human CD4+ T cells for detailed proteomic analysis, and quantitate approximately 9000 proteins across multiple donors on a dynamic background of T cell activation. Amongst 650 HIV-dependent changes (q < 0.05), we describe novel Vif-dependent targets FMR1 and DPH7, and 192 proteins not identified and/or regulated in T cell lines, such as ARID5A and PTPN22. We therefore provide a high-coverage functional proteomic atlas of HIV infection, and a mechanistic account of host factors subverted by the virus in its natural target cell.
病毒操纵宿主细胞以增强其复制能力，而鉴定被病毒靶向的细胞因子已深入了解病毒发病机制和细胞生物学。在这项研究中，我们开发了一种带有表面结合亲和素标签的 HIV 报告病毒（HIV-AFMACS），可通过链霉亲和素偶联的磁珠进行一步简单选择。我们使用该系统获得了纯的 HIV 感染的原代人 CD4+ T 细胞群体，用于在 T 细胞激活的动态背景下进行详细的蛋白质组学分析，并对多个供体进行了约 9000 种蛋白质的定量分析。在 650 个 HIV 依赖性变化（q < 0.05）中，我们描述了新型 Vif 依赖性靶标 FMR1 和 DPH7，以及 192 种在 T 细胞系中未被识别和/或调控的蛋白质，如 ARID5A 和 PTPN22。因此，我们提供了 HIV 感染的高覆盖功能蛋白质组图谱，以及病毒在其自然靶细胞中颠覆宿主因子的机制解释。
