CAJ | 학술논문

目的探讨肠道病毒71型(EV71)编码的3C蛋白酶对于肿瘤坏死因子受体相关因子3(TRAF3)的切割作用.方法采用Western blot的方法检测EV71感染对于TRAF3蛋白表达的影响.体外转染TRAF3和3C蛋白,通过Western blot检测3C对于TRAF3蛋白的影响.通过构建3C突变体,检测其蛋白酶活性对于切割的影响.通过免疫共沉淀检测3C对于TRAF3和TRIF相互作用的影响.结果 EV71感染细胞后可引起TRAF3蛋白的减少,其3C蛋白酶可以引起TRAF3的切割,蛋白酶活性位点突变后则不引起TRAF3的切割.位点突变试验显示TRAF3的Q242位突变后则不能被3C切割,且3C可以减少TRAF3与TRIF的结合.结论 EV71 3C蛋白酶可通过切割TRAF3机制在天然免疫逃逸中发挥作用.
목적 탐토장도병독71형(EV71)편마적3C단백매대우종류배사인자수체상관인자3(TRAF3)적절할작용.방법 채용Western blot적방법검측EV71감염대우TRAF3단백표체적영향.체외전염TRAF3화3C단백,통과Western blot검측3C대우TRAF3단백적영향.통과구건3C돌변체,검측기단백매활성대우절할적영향.통과면역공침정검측3C대우TRAF3화TRIF상호작용 적영향.결과 EV71감염세포후가인기TRAF3단백적감소,기3C단백매가이인기TRAF3적절할,단백매활성위점돌변후칙불인기TRAF3적절할.위점돌변시험현시TRAF3적Q242위돌변후칙불능피3C절할,차3C가이감소TRAF3여TRIF적결합.결론 EV71 3C단백매가통과절할TRAF3궤제재천연면역도일중발휘작용.
Objective To clarify whether Enterovirus 71 3C protease,a key player in immunity evasion in EV71 infection,can cleave TNF receptor-associated factor 3 (TRAF3).Methods Western blot was used to explore the TRAF3 expression in EV71-infected cells.Plasmids expressing TRAF3 and 3C were then cotransfected into cells to detect whether 3C induces TRAF3 cleavage.A mutational analysis was carried out to explore the sites in TRAF3 for 3C cleavage.The effect of 3C on the interaction between TRAF3 and TRIF was analyzed by immunoprecipitation analysis.Results The TRAF3 expression was reduced by EV71 infection,and 3C induced the cleavage of TRAF3.The cleavage depended on 3C protease activity.The Q242-G243 is the cleavage site of 3C in TRAF3.3C also impeded the interaction between TRAF3 and TRIF.Conclusions 3C induces TRAF3 cleavage in a protease activity dependent manner,which may in turn antagonizes innate immunity.