日本脑炎病毒感染宿主后m^(6)A甲基化相关蛋白的表达和定位研究  

Expression and localization of m^(6)A methylation related proteins in Japanese encephalitis virus infected hosts

作　　者：欧宇达 李晓晗 陈婧[1] 赵炳枢 陈健芃 毛玎懿 周江飞 周斌[1] OU Yuda;LI Xiaohan;CHEN Jing;ZHAO Bingshu;CHEN Jianpeng;MAO Dingyi;ZHOU Jiangfei;ZHOU Bin(College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学动物医学院,江苏南京210095

出　　处：《畜牧与兽医》2024年第2期62-68,共7页Animal Husbandry & Veterinary Medicine

基　　金：国家重点研发计划项目(2022YFD1800100)。

摘　　要：为了研究日本脑炎病毒(JEV)感染宿主后在体内外对m^(6)A甲基化相关蛋白表达和定位的影响,本试验建立了JEV感染的C57BL/6小鼠模型及乳仓鼠肾细胞(BHK-21)模型,检测JEV感染后小鼠的脑、肾及BHK-21细胞的m^(6)A修饰水平,通过蛋白免疫印迹检测小鼠脑、肾以及JEV感染不同时间点的细胞中m^(6)A相关蛋白的表达水平,共聚焦显微镜观察JEV感染细胞的m^(6)A相关蛋白的亚细胞定位变化。结果显示:与对照组相比,感染JEV后m^(6)A修饰水平极显著上升(P<0.01);感染JEV的小鼠脑、肾中m^(6)A甲基转移酶样蛋白(METTL)3表达量大幅上升,细胞中METTL3表达量随感染时间推移而逐渐增加;METTL3和METTL14的亚细胞定位发生改变,大量由细胞核转移至细胞质。综上所述,JEV可使体内外的m^(6)A水平和METTL3的表达水平上升,细胞中METTL3和METTL14的亚细胞定位改变,表明JEV的感染与m^(6)A修饰具有较强的关联性,为进一步研究m^(6)A及其相关蛋白调控JEV复制的机制奠定了基础。To investigate the effects of Japanese encephalitis virus(JEV)infection on the expression and localization of m^(6)A methylation related proteins in vivo and in vitro,this experiment established a C57BL/6 mouse model infected with JEV.JEV was infected with baby hamster kidney cells(BHK-21),and the m^(6)A modification levels in the brain,kidneys,and BHK-21 cells of mice infected with JEV were detected.Western blot was used to detect the expression levels of m^(6)A related proteins in the brain and kidneys of the mice,and in the cells at different time points.JEV infection was observed under confocal microscopy,and the subcellular localization changes of m^(6)A related proteins in the JEV infected cells were observed.The results showed that,compared with the control group,the m^(6)A modification level significantly increased after JEV infection in the treated mice(P<0.01);the expression level of m^(6)A methyltransferase like protein(METTL)3 in the brain and kidneys of the JEV infected mice rose substantially;the expression level of METTL3 in cells gradually increased with the passage of infection time;and the subcellular localization of METTL3 and METTL14 has changed,with a significant amount of transfer from the nucleus to the cytoplasm.In summary,infection of JEV promoted the expression of m^(6)A and METTL3 in vitro and in vivo,and changed the subcellular localization of METTL3 and METTL14 in cells.All this indicated a strong correlation between JEV infection and m^(6)A modification,which laid a foundation for further research on the mechanism of m^(6)A and its related proteins regulating JEV replication.

关 键 词：日本脑炎病毒 m^(6)A甲基化 甲基转移酶样蛋白3 亚细胞定位 

分 类 号：S852[农业科学—基础兽医学]