Affinity-purification of V5-tagged VZV ORFs in SK-N-BE2 cells

The alpha-herpesvirus Varicella-Zoster Virus (VZV), the causative agent of chicken pox, infects most humans worldwide and remains latent in peripheral neuronal cells for life. Its reactivation triggers herpes zoster and sporadic disseminated infection, in particular in the central nervous system (CNS) where it is associated with severe pathologies. VZV replication is efficiently controlled with antivirals, however the emergence of treatment-resistant VZV variants constitutes an increasing healthcare issue, calling for in-depth analysis of molecular VZV-host interactions to foster general knowledge and promote identification of novel therapeutic targets. Here we conducted a mass spectrometry-based survey in neuronal SK-N-BE2 cells, combining proteome analysis of VZV-infected cells with systematic characterization of VZV proteins’ functions through identifying their individual host partners and effects on the host proteome. To generate this specific dataset, we used AP-MS and a Bayesian modelling approach to characterize the VZV-host interactome. We collectively identified 1184 interactions between 56 viral and 900 human proteins.

α疱疹病毒（alpha-herpesvirus）水痘-带状疱疹病毒（Varicella-Zoster Virus, VZV）是水痘的病原体，可感染全球绝大多数人类，并终身潜伏于外周神经元细胞中。其再激活可引发带状疱疹及散发性播散性感染，尤其在中枢神经系统（central nervous system, CNS）内可引发严重病理损伤。目前抗病毒药物可有效控制VZV复制，但耐药性VZV毒株的出现正日益成为突出的医疗卫生难题，亟需深入解析VZV与宿主的分子互作机制，以深化相关理论认知并推动新型治疗靶点的发掘。本研究以神经元SK-N-BE2细胞为模型开展基于质谱的系统性研究，通过鉴定VZV病毒蛋白各自的宿主互作伴侣及其对宿主蛋白质组的影响，并结合VZV感染细胞的蛋白质组分析，全面表征VZV病毒蛋白的功能。为构建本专属数据集，我们采用亲和纯化质谱（AP-MS）结合贝叶斯建模方法，对VZV-宿主互作组进行系统表征。本研究共鉴定得到56种病毒蛋白与900种人类蛋白之间的1184组互作关系。