HMCES protein promotes the replication of porcine epidemic diarrhea virus in Vero cells

Abstract:

[Objective] To identify the key host protein that can regulate the replication of porcine epidemic diarrhea virus (PEDV). [Methods] We usedliquid chromatography with tandem mass spectrometry (LC-MS/MS) technology combined with tandem mass tag (TMT) to analyze the proteomic differences between PEDV-infected Vero cells and the uninfected group at 36 h post inoculation. A total of 114 significantly differentially expressed proteins were identified, of which 5-hydroxymethylcytosine binding, ES cell-specific protein (HMCES) was significantly up-regulated. The eukaryotic expression plasmid of HMCES was further constructed, and the effect of HMCES overexpression on PEDV replication was examined by Western blotting and real-time fluorescent quantitative PCR. The specific siRNA against HMCES gene was synthesized, and Western blotting and RT-qPCR were employed to detect the effect of siRNA on HMCES expression and the effect of interfered HMCES on PEDV replication. [Results] Overexpression of HMCES significantly promoted PEDV replication in Vero cells, and the level of replication increased in a dose-dependent manner. siRNA-341 down-regulated the expression of endogenous HMCES to inhibit PEDV replication. [Conclusion] HMCES promotes PEDV replication in Vero cells. This study provides a reference for further exploring the role and mechanism of HMCES in the anti-PEDV immune response.

Key words:HMCES;porcine epidemic diarrhea virus;virus replication;Vero cells