P58IPK通过PERK/eIF2α通路调控猪圆环病毒2型所诱导的细胞自噬

doi:10.11843/j.issn.0366-6964.2017.11.014

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (11): 2125-2134.doi: 10.11843/j.issn.0366-6964.2017.11.014

P58IPK通过PERK/eIF2α通路调控猪圆环病毒2型所诱导的细胞自噬

杨了寒¹, 王凯¹, 许苏童¹, 张敏¹, 胡林¹, 何启盖², 张淑君^1*

1. 华中农业大学动物科学技术学院农业动物遗传育种与繁殖教育部重点实验室, 武汉 430070;
2. 华中农业大学动物医学院农业微生物学国家重点实验室, 武汉 430070

收稿日期:2017-05-13 出版日期:2017-11-23 发布日期:2017-11-23
通讯作者: 张淑君,教授,E-mail:sjxiaozhang@mail.hzau.edu.cn
作者简介:杨了寒(1992-),女,江西南昌人,硕士,主要从事病毒感染与内质网应激及细胞自噬等相关研究,E-mail:hannaheeo@webmail.hzau.edu.cn
基金资助:
国家自然科学基金（31572367；31272427）；中央高校基本科研业务费

P58IPK Regulates Porcine Circovirus Type 2-induced Autophagy through PERK/eIF2α Pathway

YANG Liao-han¹, WANG Kai¹, XU Su-tong¹, ZHANG Min¹, HU Lin¹, HE Qi-gai², ZHANG Shu-jun^1*

1. Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China;
2. The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China

Received:2017-05-13 Online:2017-11-23 Published:2017-11-23

摘要/Abstract

摘要：

旨在研究猪圆环病毒2型（PCV2）感染激活的PERK/eIF2α通路是否与细胞自噬相关，以及过表达分子伴侣蛋白P58IPK在其中的作用。首先通过PERK特异性抑制剂GSK2606414和siRNA及蛋白免疫印迹检测磷酸化eIF2α（p-eIF2α）、LC3-Ⅱ和ATG5-ATG12蛋白表达，研究PCV2感染PK-15细胞24 h后PERK/eIF2α通路与细胞自噬的关系；然后通过构建和瞬时转染P58IPK真核过表达质粒及蛋白免疫印迹研究过表达P58IPK对PCV2感染PK-15诱导的PERK/eIF2α通路及细胞自噬的影响。结果显示，PERK特异性抑制剂GSK2606414和siRNA干扰PERK可极显著抑制PCV2诱导的p-eIF2α（P<0.01）和LC3-Ⅱ（P<0.01）表达；而过表达P58IPK同样可以极显著下调PCV2诱导的p-eIF2α（P<0.01）、ATG5-ATG12（P<0.01）及LC3-Ⅱ（P<0.01），且能极显著下调PCV2拷贝数（P<0.01）。结果表明，PCV2感染PK-15通过激活PERK/eIF2α通路诱导细胞自噬，过表达P58IPK通过抑制PERK通路抑制PCV2诱导的细胞自噬，从而抑制病毒复制。

Abstract:

This experiment was conducted to study the relationship between porcine circovirus type 2 (PCV2) infection activated PERK/eIF2α pathway and autophagy, and the possible function of overexpressing chaperone P58IPK within it. PK-15 cells were treated with PERK specific inhibitor GSK2606414, siRNA, followed by PCV2 infection for 24 h. Western blot was taken to measure phosphorylated eIF2α (p-eIF2α), LC3-Ⅱ and ATG5-ATG12 expression at first. Then P58IPK overexpression vector was constructed and transiently transfected, combined with Western blot to study the influence of overexpressing P58IPK on PCV2-induced PERK/eIF2α pathway and autophagy. Western blot results showed PERK specific inhibitor GSK2606414 and siRNA significantly suppressed PCV2-induced p-eIF2α (P<0.01) and LC3-Ⅱ (P<0.01) expression. While overexpressing P58IPK not only resulted in PCV2-induced p-eIF2α (P<0.01), ATG5-ATG12 (P<0.01) and LC3-Ⅱ (P<0.01) decrease, but also significantly downregulated activating PCV2 copies number (P<0.01). Results indicate PCV2 infection induces autophagy through activating PERK/eIF2α pathway. Overexpression of P58IPK represses PCV2-induced autophagy through inhibiting PERK pathway, consequently, inhibiting PCV2 replication.

中图分类号:

S852.659.2

杨了寒, 王凯, 许苏童, 张敏, 胡林, 何启盖, 张淑君. P58IPK通过PERK/eIF2α通路调控猪圆环病毒2型所诱导的细胞自噬[J]. 畜牧兽医学报, 2017, 48(11): 2125-2134.

YANG Liao-han, WANG Kai, XU Su-tong, ZHANG Min, HU Lin, HE Qi-gai, ZHANG Shu-jun. P58IPK Regulates Porcine Circovirus Type 2-induced Autophagy through PERK/eIF2α Pathway[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(11): 2125-2134.

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P58IPK通过PERK/eIF2α通路调控猪圆环病毒2型所诱导的细胞自噬

P58IPK Regulates Porcine Circovirus Type 2-induced Autophagy through PERK/eIF2α Pathway

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