Ubiquitination Of CGAS By TRAF6 Regulates Anti-DNA Viral Innate Immune Responses

Objective:To explore tumor necrosis factor receptor-associated factor 6(TRAF6)regulates anti-DNA viral innate immune response via interacting with cyclic GMP-AMP(cGAMP)synthase(cGAS).To explore the detailed mechanism,which provide a new therapeutic target for diseases caused by DNA virus infection and contribute to promoting the research from bench to bedside.Methods:(1)Plasmid co-transfection,immunoprecipitation and Western Blot were used to analyze the results,so that we explore the interaction between TRAF6 and cGAS.(2)Cells were transfected with indicated plasmids and IFN? luciferase activity was measured.And then we explore the function of TRAF6 in innate immunity.(3)The ubiquitination of cGAS by TRAF6 were detected by denatured immunoprecipitation assay.(4)Western Blot assay was used to detect the changes of protein phosphorylation in cGAS-STING signaling pathway.(5)Gene interference technique was used to knock down TRAF6 expression in cells and qPCR technique was used to detect the production of type I interferon I,the level of inflammatory factors and the replication of HSV-1 virus after virus infection and double strands DNA mimics stimulation..Results:In our study,we found that TRAF6 specifically binded to cGAS and promoted the ubiquitination of cGAS in an activity-dependent manner,thus activating the cGAS-STING signaling pathway.The levels of phosphorylated TANK-binding kinase 1(TBKl)and interferon-regulated transcription factor 3(IRF3)were increased.The production of IFN? and anti-DNA virus innate immune response were enhanced,TRAF6 knockdown or the inhibition of TRAF6 activity significantly reduced the production of IFN? and inhibit the innate immune response.Conclusion:It has been reported that TRAF6 regulated the activation of NF-kB signaling pathway through auto-ubiquitination,then affected the innate immune response.Here,we found a new mechanism of TRAF6 regulating innate immunity against DNA viruses infection.The research of cGAS is the hot topic of immunology.and the function of cGAS has been widely recognized in innate immunity and tumor immunity.We found TRAF6 is a new regulator of cGAS.Therefore,TRAF6 is likely to be involved in the process of tumor immunity,and our study provides a theoretical basis for the design of innate and tumor immune drugs.Figure[5]table[0]reference[68]...