猪GRK2蛋白抗口蹄疫病毒作用分析

doi:10.11843/j.issn.0366-6964.2023.10.031

畜牧兽医学报 ›› 2023, Vol. 54 ›› Issue (10): 4350-4361.doi: 10.11843/j.issn.0366-6964.2023.10.031

猪GRK2蛋白抗口蹄疫病毒作用分析

陈文哲¹, 张向乐², 顾峰幸², 赵振翔², 李康丽², 薛钊宁², 郑海学², 张小丽^1*, 朱紫祥^2*

1. 甘肃农业大学动物医学院, 兰州 730070;
2. 中国农业科学院兰州兽医研究所, 兰州大学动物医学与生物安全学院, 家畜疫病病原生物学国家重点实验室, 兰州 730046

收稿日期:2023-01-10 出版日期:2023-10-23 发布日期:2023-10-26
通讯作者: 张小丽,主要从事兽医微生物及免疫学研究,E-mail:zhangxiaol@gsau.edu.cn;朱紫祥,主要从事口蹄疫病毒、非洲猪瘟病毒等感染与免疫机制及疫苗设计创制研究,E-mail:zhuzixiang@caas.cn
作者简介:陈文哲(1998-),男,甘肃天水人,硕士生,主要从事天然免疫及病毒致病机制的研究,E-mail:1215156031@qq.com
基金资助:
国家重点研发计划项目（2021YFD1800300）；甘肃省重大专项（21ZD3NA001，20ZD7NA006，19ZDNA001）；"十四五"广东省农业科技创新十大主攻方向"揭榜挂帅"项目（2022SDZG02）；生猪产业体系（CARS-35）；绒毛羊产业体系（CARS-39-13）；中国农业科学院基本科研业务费（1610312022006）

Evaluation of the Antiviral Effect of Porcine GRK2 against Foot-and-mouth Disease Virus

CHEN Wenzhe¹, ZHANG Xiangle², GU Fengxing², ZHAO Zhenxiang², LI Kangli², XUE Zhaoning², ZHENG Haixue², ZHANG Xiaoli^1*, ZHU Zixiang^2*

1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;
2. State key Laboratory of Veterinary Etiological Biology, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

Received:2023-01-10 Online:2023-10-23 Published:2023-10-26

摘要/Abstract

摘要： 为研究G蛋白偶联受体激酶2（G protein-coupled receptor kinase 2，GRK2）对口蹄疫病毒（foot-and-mouth disease virus，FMDV）复制的影响，从猪的PK-15细胞中提取RNA，通过聚合酶链式反应（polymerase chain reaction，PCR）扩增出猪GRK2的完整CDS序列，并构建了猪GRK2真核表达质粒。Western blot和间接免疫荧光试验表明GRK2可表达并定位于细胞质中。FMDV感染PK-15细胞的GRK2转录水平和蛋白水平呈先上升后下降趋势；过表达GRK2可显著抑制FMDV的复制，下调表达GRK2可显著促进FMDV的复制，表明GRK2在FMDV感染过程中具有抗病毒作用。进一步研究发现FMDV 3C^pro蛋白可与GRK2互作并降解GRK2，促进FMDV的复制。研究结果为进一步分析宿主蛋白GRK2的抗FMDV的分子机制奠定了基础。

关键词: 口蹄疫病毒, G蛋白偶联受体激酶2, 抗病毒作用

Abstract: To investigate the effect of G protein-coupled receptor kinase 2 (GRK2) on the replication of foot-and-mouth disease virus (FMDV), the porcine cellular RNA from PK-15 cells were extracted and the complete CDS region of GRK2 was obtained by Polymerase Chain Reaction (PCR) methods. Subsequently, the eukaryotic expressing plasmid was constructed. The Western blot and indirect immunofluorescence experiments showed that GRK2 was expressed in the transfected cells and located in the cytoplasm. The infection assay suggested that the transcription level and protein level of GRK2 firstly increased and then decreased in FMDV-infected PK-15 cells. Overexpression assay indicated that GRK2 significantly inhibited FMDV replication, and siRNA assay showed that downregulation of GRK2 promoted FMDV replication, revealing the antiviral function of GRK2 against FMDV. Further studies showed that FMDV 3C^pro protein interacted with GRK2 and degraded GRK2 to promote FMDV replication. In conclusion, this study laid a foundation for analysis of the antiviral mechanism of host protein GRK2 against FMDV.

Key words: foot-and-mouth disease virus, G protein-coupled receptor kinase 2, antiviral effect

中图分类号:

S852.659.6

陈文哲, 张向乐, 顾峰幸, 赵振翔, 李康丽, 薛钊宁, 郑海学, 张小丽, 朱紫祥. 猪GRK2蛋白抗口蹄疫病毒作用分析[J]. 畜牧兽医学报, 2023, 54(10): 4350-4361.

CHEN Wenzhe, ZHANG Xiangle, GU Fengxing, ZHAO Zhenxiang, LI Kangli, XUE Zhaoning, ZHENG Haixue, ZHANG Xiaoli, ZHU Zixiang. Evaluation of the Antiviral Effect of Porcine GRK2 against Foot-and-mouth Disease Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4350-4361.

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猪GRK2蛋白抗口蹄疫病毒作用分析

Evaluation of the Antiviral Effect of Porcine GRK2 against Foot-and-mouth Disease Virus

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