Black carp IKKε collaborates with IRF3 in the antiviral signaling.
Black carp IKKε collaborates with IRF3 in the antiviral signaling.
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DOI：
10.1016/j.fsi.2021.08.034
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发表时间：
2021-09
影响因子：
4.7
通讯作者：
Chanyuan Wang;Jun Li;Xiao Yang;Qun Wang;Huijuan Zhong;Yankai Liu;Weiyi Yan;Yunfan He;
中科院分区：
文献类型：
--
作者：
Chanyuan Wang;Jun Li;Xiao Yang;Qun Wang;Huijuan Zhong;Yankai Liu;Weiyi Yan;Yunfan He;
干扰素调节因子3（Interferon Regulatory Factor 3，IRF 3）由IκB激酶ε（I κ K ε）和Tank-结合激酶1（Tank-binding kinase 1，TBK 1）激活，在脊椎动物干扰素信号转导中起重要作用。然而，IKKε对硬骨鱼IRF 3的调控在很大程度上仍然未知。本研究克隆了青鱼IRF 3的同源基因（bcIRF 3），并对其进行了结构分析。bcIRF 3在宿主细胞中的转录水平在不同的刺激条件下均有所增加。bcIRF 3主要分布在胞浆区，但病毒感染后，bcIRF 3易位到细胞核内。报告基因检测表明，bcIRF 3具有诱导IFN活性，表达bcIRF 3的EPC细胞对草鱼呼肠孤病毒（GCRV）和鲤鱼春季病毒血症病毒（SVCV）均有较强的抗病毒活性。此外，bcIRF 3基因敲低降低了宿主细胞的抗病毒能力，并且bcIRF 3基因敲低的宿主细胞中抗病毒相关细胞因子的转录水平明显低于对照细胞。报告基因实验和空斑实验结果表明，bcIKKε能显著增强bcIRF 3介导的IFN产生和抗病毒活性。免疫荧光染色和免疫共沉淀实验表明bcIKKε与bcIRF 3相互作用。结果表明，bcIRF 3和bcIKK ε共表达时，两者的核转位相互促进，而蛋白水平则相互降低。因此，我们的数据支持这样的结论，即bcIKKε与bcIRF 3相互作用，并在宿主先天免疫激活过程中增强bcIRF 3介导的抗病毒信号传导。
Interferon regulatory factor 3 (IRF3) is activated by IκB kinase ε (IKKε) and Tank-binding kinase 1 (TBK1), which plays a crucial role in the interferon signaling in vertebrates. However, the regulation of teleost IRF3 by IKKε remains largely unknown. In this study, the IRF3 homologue (bcIRF3) of black carp (Mylopharyngodon piceus) has been cloned and characterized. The transcription of bcIRF3 was detected to increase in host cells in response to different stimuli. bcIRF3 distributed predominantly in the cytosolic area; however, translocated into nuclei after virus infection. bcIRF3 showed IFN-inducing ability in reporter assay and EPC cells expressing bcIRF3 showed enhanced antiviral ability against both grass carp reovirus (GCRV) and spring viremia of carp virus (SVCV). Moreover, knockdown of bcIRF3 reduced the antiviral ability of the host cells, and the transcription of antiviral-related cytokines was obviously lower in bcIRF3-deficient host cells than that of control cells. The data of reporter assay and plaque assay demonstrated that bcIKKε obviously enhanced bcIRF3-mediated IFN production and antiviral activity. Immunofluorescent staining and co-immunoprecipitation assay revealed that bcIKKε interacted with bcIRF3. It was interesting that the nuclear translocation of bcIRF3 and bcIKKε was enhanced by each other when these two molecules were co-expressed in the cells, however, the protein levels of bcIRF3 and bcIKKε were decreased mutually. Thus, our data support the conclusion that bcIKKε interacts with bcIRF3 and enhances bcIRF3-mediated antiviral signaling during host innate immune activation.