ZWINT在RLR抗病毒先天免疫信号通路中的作用

Effects of ZWINT on RLR antiviral innate immune signaling

目的探究ZWINT(ZW10 interacting kinetochore protein)对TBK1(TANK-binding kinase 1)或仙台病毒(Sendai virus,SeV)诱导的RLR(RIG-I like receptor)抗病毒先天免疫信号通路的影响。方法应用免疫共沉淀的方法检测ZWINT与RLR信号通路分子TBK1、VISA、RIG-I之间的相互作用;用非变性聚丙烯酰胺凝胶电泳实验验证ZWINT对TBK1或SeV诱导的IRF3二聚化的作用;利用双荧光素酶报告基因实验检测TBK1或SeV介导下ZWINT对干扰素启动子IFN-β的激活效果;通过实时荧光定量PCR实验检测ZWINT对SeV诱导的IFN-β转录的影响。结果 ZWINT与TBK1、VISA和RIG-I都有相互作用,过表达ZWINT增强了TBK1或SeV诱导的IRF3的二聚化水平以及IFN-β启动子的活性,此外,过表达ZWINT也加强了SeV诱导的IFN-β的转录。结论ZWINT是RLR抗病毒先天免疫信号通路中的正调控因子。

This study was performed to explore the effect of ZWINT on RLR antiviral innate immune signaling pathway induced by TBK1 or SeV.The interaction between ZWINT and RLR signaling pathway molecules TBK1, VISA and RIG-I was detected by co-immunoprecipitation;the effect of ZWINT on TBK1 or SeV-triggered IRF3 dimerization was confirmed by non-denatured polyacrylamide gel electrophoresis experiment.Furthermore,the activated effect of ZWINT on interferon promoter IFN-β induced by TBK1 or SeV was detected by dual luciferase reporter gene assay;the effect of ZWINT on IFN-β transcription induced by SeV was analyzed by real-time PCR assay.Data showed that ZWINT interacted with TBK1,VISA and RIG-I.Overexpression of ZWINT enhanced the dimerization of IRF3 induced by TBK1 or SeV,and also promoted the activity of IFN-β promoter.In addition, overexpression of ZWINT also reinforced the transcription of IFN-β triggered by SeV.In conclusion,ZWINT is a positive regulator of the RLR antiviral innate immune signaling pathway.