CVA16感染对m^6A甲基化相关蛋白表达和定位的影响  

Effects of CVA16 infection on m^6A methylation-related protein expression and localization

作　　者：李卫宇 蒋曦 郑雪麟 朱文兵[1] 刘卓航 李慧[1] 张雪梅[1] 吴忠香[1] 宋杰[1] 董少忠[1] Li Weiyu;Jiang Xi;Zheng Xuelin;Zhu Wenbing;Liu Zhuohang;Li Hui;Zhang Xuemei;Wu Zhongxiang;Song Jie;Dong Shaozhong(Yunnan Key Laboratory of Vaccine Research and Development on Severe Infectious Disease,Institute of Medical Biology,Chinese Academy of Medical Sciences&Peking Union Medical College,Kunming 650118,China)

机构地区：[1]中国医学科学院北京协和医学院,医学生物学研究所,云南省重大传染病疫苗研发重点实验室,昆明650118

出　　处：《中华微生物学和免疫学杂志》2020年第7期530-537,共8页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金项目(31700153);中央高校基本科研业务费(3332019004)。

摘　　要：目的本研究旨在探究柯萨奇病毒A组16型(Coxsackievirus A 16,CVA16)感染后是否会影响N6-甲基腺嘌呤(N6-methyladenosine,m6A)甲基化相关蛋白在人呼吸道上皮细胞(16HBE)、ICR乳鼠和SCARB2人源化小鼠中的表达,以及在细胞中的定位。方法病毒分别以感染复数(MOI)=0.1感染16HBE和107 CCID50/ml感染小鼠,Western blot分析甲基转移酶、去甲基化酶和甲基化阅读蛋白的表达变化,免疫荧光观察这些蛋白质在细胞中的定位。结果研究结果发现,随着CVA16病毒感染时间的增加,m6A甲基化相关蛋白在细胞中表达水平逐渐下调,在ICR乳鼠和SCARB2小鼠中无明显变化;病毒感染后,m6A甲基化相关蛋白在细胞核与细胞质中重新分布,甚至发生降解。结论CVA16在宿主细胞中复制时可以改变m6A甲基化修饰相关蛋白的表达和细胞定位。本研究结果提示m6A修饰可能是肠道病毒治疗的潜在新靶点。Objective To investigate whether coxsackievirus A 16(CVA16)infection would affect the expression of N6-methyladenosine(m6A)methylation-related proteins in human bronchial epithelial cells(16HBE),ICR suckling mice and SCRBA2 humanized mice and influence their subcellular localization.Methods CVA16 was used to infect 16HBE cells at a multiplicity of infection(MOI)of 0.1 and mice at 107 CCID50/ml.Changes in the expression of methyltransferases,demethylases and methylated reading proteins were analyzed by Western blot.Cellular localization of these proteins was observed using immunofluorescence.Results The expression of m6A methylation-related proteins was gradually reduced in CVA16-infected cells with time,but showed no obvious change in ICR suckling mice or SCRBA2 humanized mice.After infection,m6A methylation-related proteins were redistributed in both the nucleus and cytoplasm and even degraded.Conclusions CVA16 replication in host cells altered the expression and cellular localization of m6A methylation-related proteins,which indicated that m6A modification might be a new potential target for enterovirus therapy.

关 键 词：柯萨奇病毒A组16型(CVA16) m^6A甲基化相关蛋白 

分 类 号：R373.23[医药卫生—病原生物学]