Pseudorabies virus EP0 recruits TECPR2 and CK2 to promote COPII accumulation and viral egress.

The rapid assembly of alpha-herpesvirus imposes a substantial metabolic burden on the host secretory pathway, requiring efficient ER-to-Golgi transport of viral glycoproteins. However, the mechanisms by which viruses remodel ER exit sites (ERES) to accommodate this surge in cargo remain poorly understood. Here, we report that Pseudorabies virus (PRV) exploits the host autophagy regulator TECPR2 (Tectonin beta-propeller repeat containing 2) to orchestrate COPII (Coat protein complex II) component accumulation and facilitate viral egress. We demonstrate that the viral immediate-early protein EP0, interacts with the WD40 domain of TECPR2 in the cytoplasm during late infection. This interaction recruits the host kinase Casein Kinase 2 (CK2) to form a cascade that stabilizes both EP0 and the COPII outer coat protein SEC31A. Consequently, this axis enhances the secretory capacity of infected cells in a manner dependent on CK2 activity but independent of transcriptional upregulation. It is worth noting that under this pathway, 25-hydroxy cholesterol (25-HC) can inhibit viral transmission, which disrupts the EP0-TECPR2-CK2 axis and suppresses the accumulation of COPII components. In vivo, 25-HC treatment significantly reduced viral loads and attenuated neurovirulence in a murine infection model. These findings reveal how PRV utilizes autophagy factors to promote secretory pathways, and highlight the EP0-TECPR2-CK2 axis, which is cholesterol-dependent, as a potential therapeutic target.