Whole-cell and plasma membrane proteomics of herpes simplex virus-1 infection reveals cell surface remodelling via pUL56-mediated GOPC degradation

Herpesviruses are ubiquitous in the human population and they extensively remodel the cellular environment during infection. Multiplexed quantitative proteomic analysis over the time-course of herpes simplex virus (HSV)-1 infection was used to characterize changes in the host-cell proteome and the kinetics of viral protein production. Several host-cell proteins are targeted for rapid degradation by HSV-1, including the cellular trafficking factor GOPC. We show that the poorly-characterized HSV-1 pUL56 directly binds GOPC, stimulating its ubiquitination and proteasomal degradation. Plasma membrane profiling revealed that pUL56 mediates specific changes to the cell surface proteome of infected cells, including loss of IL18 receptor and Toll-like receptor 2, and that cell surface expression of Toll-like receptor 2 is GOPC-dependent. Our study provides significant resources for future investigation of HSV-host interactions and highlights an unanticipated and efficient mechanism whereby a single virus protein targets a cellular trafficking factor to modify the surface of infected cells.

This deposition includes quantified peptide data for the five experiments presented in the manuscript. In HSV_UL56_GOPC_Figures3_and_6.xlsx, 090316_Exp[123] relate to the SILAC IP shown in Figure 3, and Exp1_2018_singleshot_SILAC_LMH_Mock_WT_dUL56" and
"Exp2_2018_Fraction[123456]_SILAC_LMH_Mock_WT_dUL56" relate to the plasma membrane profiling experiment shown in Figure 6.

疱疹病毒在人类群体中广泛存在，它们在感染过程中对细胞环境进行广泛重塑。通过对单纯疱疹病毒（HSV-1）感染过程中的多重定量蛋白质组学分析，本研究旨在描绘宿主细胞蛋白质组的变化以及病毒蛋白质生产的动力学。HSV-1靶向多个宿主细胞蛋白进行快速降解，包括细胞运输因子GOPC。本研究揭示了HSV-1的未充分研究的pUL56蛋白可直接结合GOPC，激发其泛素化和蛋白酶体降解。血浆膜分析显示，pUL56介导了感染细胞细胞表面蛋白质组的特定变化，包括IL18受体和Toll样受体2的丢失，且Toll样受体2的细胞表面表达依赖于GOPC。本研究为未来HSV与宿主相互作用的研究提供了重要的资源，并突显了一种出乎意料且高效的机制，即单个病毒蛋白靶向细胞运输因子以改变感染细胞的表面特性。本沉积包括论文中展示的五项实验的量化肽数据。在HSV_UL56_GOPC_Figures3_and_6.xlsx中，090316_Exp[123]与图3中所示的SILAC IP相关，而“Exp1_2018_singleshot_SILAC_LMH_Mock_WT_dUL56”和“Exp2_2018_Fraction[123456]_SILAC_LMH_Mock_WT_dUL56”与图6中所示的血浆膜分析实验相关。