甲基转移酶样蛋白3及去甲基化酶肥胖相关蛋白对H7N9病毒复制的影响及转录组学分析  

Effects of methyltransferase like 3 and fat mass and obesity-associated protein on H7N9 virus replication and transcriptome analysis

作　　者：罗颖 孙颖 刘慧 彭博 武伟华 王昕 雷雨璇 郑青[1] 房师松 Luo Ying;Sun Ying;Liu Hui;Peng Bo;Wu Weihua;Wang Xin;Lei Yuxuan;Zheng Qing;Fang Shisong(College of Pharmacy,Jinan University,Guangzhou 510632,China;Shenzhen Center for Disease Control and Prevention,Shenzhen 518055,China;School of Public Health,Sun Yat-sen University,Guangzhou 510080,China)

机构地区：[1]暨南大学药学院,广州510632 [2]深圳市疾病控制预防中心,518055 [3]中山大学公共卫生学院,广州510080

出　　处：《中华实验和临床病毒学杂志》2020年第4期385-390,共6页Chinese Journal of Experimental and Clinical Virology

基　　金：国家自然科学基金(81871631)。

摘　　要：目的探究甲基转移酶样蛋白3(methyltransferase like 3,METTL3)和去甲基化酶肥胖相关蛋白(fat mass and obesity-associated protein,FTO)在H7N9病毒感染中发挥的作用及可能的分子机制。方法利用Western blot(WB)检测H7N9病毒感染A549细胞后METTL3和FTO的表达。敲低和过表达METTL3和FTO,H7N9病毒感染后用WB和TCID50检测其对病毒复制的影响。利用转录组测序方法对METTL3或FTO敲低及野生型对照细胞进行了转录组水平基因差异表达分析,对发挥作用的潜在靶标进行初步分析。结果H7N9病毒感染A549细胞24 h后METTL3的表达上调,FTO无明显变化。敲低METTL3和FTO后,病毒核蛋白(nucleoprotein,NP)表达水平和病毒滴度显著下降,过表达METTL3和FTO后,病毒滴度显著增加。转录组测序结果显示在METTL3或FTO敲低组与对照组之间分别发现314和555个差异表达基因,GO功能富集分析和KEGG通路富集分析结果表明这些基因与宿主免疫应答相关。结论METTL3和FTO可能通过调节宿主-病毒相互作用在H7N9病毒感染中起关键作用。Objective To investigate the role and possible molecular mechanisms of m6A methyltransferase METTL3 and demethylase FTO in H7N9 virus infection.Methods Western blot(WB)was used to detect the expression of METTL3 and FTO after H7N9 virus infected A549 cells.After knocking down and over-expressing METTL3 and FTO,H7N9 virus infection was tested by WB and TCID50 for its effect on virus replication.Transcriptome sequencing method were used to analyze differentially expressed genes at the transcriptome level in METTL3 or FTO knockdown and wild-type control cells,and preliminary analysis of potential targets was performed.Results After 24 h of H7N9 virus infection in A549 cells,METTL3 expression was up-regulated,and FTO did not significantly change.After knocking down METTL3 and FTO,the virus nucleoprotein(NP)expression level and virus titer decreased significantly.Consistent with the result of the knockdown experiments,we found that the viral titer was increased by METTL3 and FTO overexpression.Transcriptome sequencing result showed that 314 and 555 differentially expressed genes were found between the METTL3 or FTO knockdown group and the control group,respectively.GO functional enrichment analysis and KEGG pathway enrichment analysis showed that these genes were related to the host immune response.Conclusions METTL3 and FTO may play a key role in H7N9 virus infection by regulating host-virus interactions.

关 键 词：METTL3 FTO H7N9 转录组分析 

分 类 号：R373.13[医药卫生—病原生物学]