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. We identified host proteins and signalling routes involved in VZV-mediated cell cycle progression, blockade of apoptosis, alteration of neuronal differentiation and innate immunity evasion. We functionally evaluated the 116 most prominent host proteins by loss-of-function assay and unveiled yet uncharacterized dependency and restriction factors. We generated stable knockout BFP positive SK-N-BE2 cells to validate their function in VZV replication assays as compared to Non-Targeting-Control (NTC) cells.