Receptor-interacting serine/threonine protein kinase 1 (RIPK1) is an important regulator of necroptosis and involved in innate immune response in human and mammal; however, its function in teleost fish mains largely unknown. In this paper, the RIPK1 homologue of black carp (Mylopharyngodon piceus) has been cloned and characterized to explore its role in immunity. Black carp RIPK1 (bcRIPK1) possesses the similar structure to its mammalian counterpart, which has been identified as a cytosolic protein by immunofluorescence staining. Overexpressed bcRIPK1 in host cells led to the decreased transcription of interferon (IFN) and interferon stimulated genes, and exogenous bcRIPK1 in EPC cells led to the decreased transcription of interferon promoters in reporter assay. Our previous study has identified that black carp MAVS (bcMAVS) functions as an antiviral adaptor protein against both grass carp reovirus (GCRV) and spring viremia of carp virus (SVCV). The reporter assay showed that the IFN-inducing ability of bcMAVS was dampened by bcRIPK1 and the plaque assay demonstrated that the antiviral activity of bcMAVS was inhibited by bcRIPK1. The immunofluorescent staining and co-immunoprecipitation identified the interaction between these two molecules. Thus, the data generated in this paper support the conclusion that bcRIPK1 interacts with bcMAVS and negatively regulates bcMAVS-mediated antiviral signaling.

中文翻译：

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黑鲤鱼 RIPK1 在先天免疫激活过程中负向调节 MAVS 介导的抗病毒信号。

受体相互作用的丝氨酸/苏氨酸蛋白激酶 1 (RIPK1) 是坏死性凋亡的重要调节因子，参与人类和哺乳动物的先天免疫反应；然而，它在硬骨鱼干线中的功能很大程度上未知。在本文中，已经克隆并表征了青鱼（Mylopharyngodon piceus）的 RIPK1 同源物，以探索其在免疫中的作用。黑鲤 RIPK1 (bcRIPK1) 具有与其哺乳动物对应物相似的结构，已通过免疫荧光染色鉴定为胞质蛋白。宿主细胞中过表达的 bcRIPK1 导致干扰素 (IFN) 和干扰素刺激基因的转录减少，EPC 细胞中的外源 bcRIPK1 导致报告基因检测中干扰素启动子的转录减少。我们之前的研究已经确定，青鱼 MAVS (bcMAVS) 可作为抗草鱼呼肠孤病毒 (GCRV) 和春季鲤病毒血症病毒 (SVCV) 的抗病毒衔接蛋白发挥作用。报告基因检测表明 bcMAVS 的 IFN 诱导能力被 bcRIPK1 抑制，斑块检测表明 bcMAVS 的抗病毒活性被 bcRIPK1 抑制。免疫荧光染色和免疫共沉淀确定了这两种分子之间的相互作用。因此，本文产生的数据支持 bcRIPK1 与 bcMAVS 相互作用并负调节 bcMAVS 介导的抗病毒信号传导的结论。报告基因检测表明 bcMAVS 的 IFN 诱导能力被 bcRIPK1 抑制，斑块检测表明 bcMAVS 的抗病毒活性被 bcRIPK1 抑制。免疫荧光染色和免疫共沉淀确定了这两种分子之间的相互作用。因此，本文产生的数据支持 bcRIPK1 与 bcMAVS 相互作用并负调节 bcMAVS 介导的抗病毒信号传导的结论。报告基因检测表明 bcMAVS 的 IFN 诱导能力被 bcRIPK1 抑制，斑块检测表明 bcMAVS 的抗病毒活性被 bcRIPK1 抑制。免疫荧光染色和免疫共沉淀确定了这两种分子之间的相互作用。因此，本文产生的数据支持 bcRIPK1 与 bcMAVS 相互作用并负调节 bcMAVS 介导的抗病毒信号传导的结论。