Key Laboratory of Applied Technology On Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Research Center for Animal Health Diagnostics & Advanced Technology, Zhejiang International Science and Technology Cooperation Base for Veterinary Medicine and Health Management, China-Australia Joint Laboratory for Animal Health Big Data Analytics, College of Animal Science and Technology & College of Veterinary Medicine of Zhejiang A&F University, 666 Wusu Street, Lin'an District, Hangzhou, 311300, Zhejiang, China.
Vet Res. 2023 Feb 3;54(1):9. doi: 10.1186/s13567-023-01139-z.
Of the three branches of unfolded protein response (UPR) that were reportedly activated by porcine epidemic diarrhea virus (PEDV), PERK is recently shown to act as an upstream regulator of oxidative response of the cells. However, it remains unknown if and how PERK activation during PEDV infection would result in oxidative stress, and whether activation of PERK and its downstream molecules affect PEDV replication. Here, we demonstrate that infection with the PEDV strain YJH/2015 triggered UPR in Vero E6 cells by activating the PERK/eIF2α pathway and led to significant increase in the expression of proapoptotic protein C/EBP homologous protein (CHOP) and ER oxidoreductase 1 alpha (ERO1α). Inhibition of PERK by short hairpin RNA (shRNA) or GSK2606414 and knockdown of CHOP by small interfering RNA reduced expression of ERO1α and generation of ROS in PEDV-infected cells. Inhibition of ERO1α by shRNA or EN460 decreased PEDV-induced ROS generation. Genetic or pharmacological inhibition of each component of PERK, CHOP, ERO1α, and ROS led to significant suppression of PEDV replication. Collectively, our study provides the first evidence that PEDV manipulates endoplasmic reticulum to perturb its redox homeostasis via the PERK-CHOP-ERO1α-ROS axis in favor of its replication.
在据称被猪流行性腹泻病毒 (PEDV) 激活的未折叠蛋白反应 (UPR) 的三个分支中，PERK 最近被证明是细胞氧化反应的上游调节剂。然而，尚不清楚 PERK 在 PEDV 感染期间的激活是否会导致氧化应激，以及 PERK 的激活及其下游分子是否会影响 PEDV 的复制。在这里，我们证明了感染 PEDV 株 YJH/2015 通过激活 PERK/eIF2α 途径在 Vero E6 细胞中引发 UPR，并导致促凋亡蛋白 C/EBP 同源蛋白 (CHOP) 和 ER 氧化还原酶 1α (ERO1α) 的表达显著增加。短发夹 RNA (shRNA) 或 GSK2606414 抑制 PERK 或小干扰 RNA 敲低 CHOP 可降低 PEDV 感染细胞中 ERO1α 的表达和 ROS 的产生。shRNA 或 EN460 抑制 ERO1α 可降低 PEDV 诱导的 ROS 产生。PERK、CHOP、ERO1α 和 ROS 的每个成分的遗传或药理学抑制都导致 PEDV 复制的显著抑制。总之，我们的研究首次提供了证据，证明 PEDV 通过 PERK-CHOP-ERO1α-ROS 轴操纵内质网以破坏其氧化还原稳态，从而有利于其复制。
