PEDV hijacks DNA damage pathways and manipulates the cell cycle to promote self-replication

Abstract:

[Objective] To study the effect of DNA damage response (DDR) on the replication of porcine epidemic diarrhea virus (PEDV). [Methods] Specific inhibitors were used to detect whether DDR pathway was involved in PEDV replication. the comet assay was employed to observe the DNA damage caused by PEDV infection in Vero cells. the changes in the expression levels of proteins in the DDR pathway and cell cycle of PEDV-infected Vero cells were determined by Western blotting and flow cytometry, respectively. [Results] The ATM inhibitor KU55933 significantly inhibited the replication of PEDV, with the virus titer decreasing from (5.50±0.25) log₁₀ TCID₅₀/mL to (3.15±0.15) log₁₀ TCID₅₀/mL. PEDV infection caused DNA damage in Vero cells during 12–60 h. ATM, ATR, Chk1, Chk2, and p53 were activated by PEDV infection of Vero cells. Especially, p-Chk2 and p-p53 showcased high expression during virus replication. In addition, PEDV infection led to the stagnation of Vero cells in the S phase. During virus replication, the expression of Cyclin B1 was first downregulated and then upregulated significantly. [Conclusion] PEDV perhaps utilized DNA damage pathway hijacks the ATM-Chk2 to manipulate the cell cycle and promote self-replication. The results provided a basis for elucidating the replication and infection mechanisms of PEDV and developing new potential antiviral targets.

Key words:porcine epidemic diarrhea virus;DNA damage;cell cycle;virus replication