热休克蛋白27对水泡性口炎病毒体外增殖的调控作用  

Regulation of Heat Shock Protein 27 on the Proliferation of Vesicular Stomatitis Virus in vitro

作　　者：李殿玉 莫荣纤 赵旭 高铭 李洪珊 白辉盛 马瑞仙 李向茸[1,3] 冯若飞[1,3] LI Dianyu;MO Rongqian;ZHAO Xu;GAO Ming;LI Hongshan;BAI Huisheng;MA Ruixian;LI Xiangrong;FENG Ruofei(Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030,China;Life Science and Engineering College of Northwest Minzu University,Lanzhou 730030,China;Engineering Research Center of Key Technology and Industrialization of Cell-based Vaccine,Ministry of Education,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030,China)

机构地区：[1]西北民族大学生物医学研究中心,生物工程与技术国家民委重点实验室,兰州730030 [2]西北民族大学,生命科学与工程学院,兰州730030 [3]西北民族大学生物医学研究中心,细胞基质疫苗关键技术与产业化教育部工程研究中心,兰州730030

出　　处：《畜牧兽医学报》2024年第6期2540-2549,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：西北民族大学中央高校基本科研业务费资金项目(31920230162,31920230163);国家自然科学基金(32260037);西北民族大学国家级大学生创新创业训练计划项目(202210742022);西北民族大学大学生创新训练项目(X202310742266)。

摘　　要：本研究旨在探究热休克蛋白27(heat shock protein 27, HSP27)对水泡性口炎病毒(vesicular stomatitis virus, VSV)体外增殖及VSV感染介导的维甲酸诱导基因I样受体家族(retinoid acid-inducible gene-I-like receptor, RLR)信号通路的作用及机制。利用实时荧光定量PCR、免疫印迹、免疫共沉淀、病毒滴度测定及免疫荧光等方法,首先检测VSV感染对HSP27 mRNA和蛋白表达的影响,进而验证过表达和干扰HSP27对VSV增殖的影响,进一步探究HSP27对VSV感染介导的RLR信号通路的影响,最后分析HSP27与RLR信号通路靶分子的相互作用及共定位情况。结果表明,VSV感染可促使HSP27基因转录和蛋白表达上调,稳定过表达和瞬时过表达HSP27均能显著抑制VSV的体外增殖;干扰宿主细胞中HSP27表达可促进VSV增殖。HSP27能够增强VSV及维甲酸诱导基因I蛋白(retinoic acid-inducible gene I protein, RIG-I)介导的IFN-β的产生及RIG-I的蛋白表达,而且HSP27与RIG-I相互作用并共定位于细胞质中。本研究揭示了HSP27靶向RIG-I上调其表达,增强RLR信号通路的转导,进而负调控VSV体外增殖,为深入揭示宿主因子HSP27在病毒感染中的作用机制奠定基础。This study was conducted to investigate the effect and mechanism of heat shock protein 27(HSP27)on the proliferation of vesicular stomatitis virus(VSV)in vitro and the retinoid acid-inducible gene-I-like receptor(RLR)signaling pathway mediated by VSV infection.Real-time quantitative PCR,Western blotting,Co-immunoprecipitation,TCID 50 assay,and immunofluorescence methods were employed to reveal the effects of VSV infection on the mRNA and protein expression of HSP27,and then the effects of overexpression and interference with HSP27 on the proliferation of VSV,and further explore the effects of HSP27 on the RLR signaling pathway mediated by VSV infection.Finally,the interaction and co-localization between HSP27 and the target molecules in RLR signaling pathway were analyzed.The results showed that VSV infection could increase the gene transcription and protein expression of HSP27,both stable and transient overexpression of HSP27 significantly inhibited the proliferation of VSV in vitro.Interfering with HSP27 expression in host cells promoted the proliferation of VSV.Further studies showed that HSP27 enhanced both VSV and retinoic acid-inducible gene I protein(RIG-I)mediated IFN-βproduction and the protein expression of RIG-I,and that HSP27 interacted with RIG-I and co-localized in the cytoplasm.This study for the revealed the molecular mechanism by which HSP27 targets RIG-I to upregulate its expression,enhance the transduction of RLR signaling pathways,and then negatively regulate the proliferation of VSV in vitro,laying a foundation for further revealing the mechanism of host factor HSP27 in viral infection.

关 键 词：热休克蛋白27 水泡性口炎病毒 RLR信号通路 维甲酸诱导基因I蛋白 

分 类 号：S852.659.5[农业科学—基础兽医学]