RIPK3 collaborates with RIPK1 to inhibit MAVS-mediated signaling during black carp antiviral innate immunity
RIPK3 collaborates with RIPK1 to inhibit MAVS-mediated signaling during black carp antiviral innate immunity
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DOI：
10.1016/j.fsi.2021.06.011
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发表时间：
2021-06-20
影响因子：
4.7
通讯作者：
Feng, Hao
中科院分区：
文献类型：
--
作者：
Dai, Yuhan;Cao, Yingyi;Feng, Hao
MAVS/IFN信号的激活和减弱对于宿主防御病毒感染至关重要，因此导致MAVS介导的信号的精细调节。然而，有关MAVS/IFN信号在硬骨鱼的调节机制还没有很好地理解。RIPK 3已被鉴定为人类和哺乳动物中坏死性凋亡、细胞凋亡和炎症信号传导的关键调节因子。在这里，我们报告的RIPK 3同源物从青鱼青鱼（bcRIPK 3）的鉴定和描述其在调节MAVS/IFN信号转导的作用。qPCR结果表明，bcRIPK 3在poly（I：C）或LPS刺激下被转录激活。免疫印迹和免疫荧光染色结果表明，bcRIPK 3是一种分子量为47 kDa的胞浆蛋白。与其哺乳动物对应物一样，bcRIPK 3在诱导细胞死亡方面表现出保守的功能。报告基因和噬斑实验表明，bcRIPK 3的过表达限制了bcMAVS激活的青鱼和斑马鱼干扰素启动子的转录，抑制了bcMAVS介导的抗病毒活性。值得注意的是，与共表达bcRIPK 3和bcMAVS的细胞相比，共表达bcRIPK 3、bcRIPK 1和bcMAVS的EPC细胞表现出明显减弱的抗病毒活性;随后的co-IP测定鉴定了bcRIPK 3和bcRIPK 1之间的相互作用。我们的研究结果首次在硬骨鱼中阐明了青鱼RIPK 3与RIPK 1相互作用抑制MAVS介导的抗病毒信号传导。
Both the activation and attenuation of MAVS/IFN signaling are critical for host defensing against viral infection and thus lead to an elaborate regulation of MAVS-mediated signaling. However, the regulatory mechanisms concerning MAVS/IFN signaling in teleost fish are not well understood. RIPK3 has been identified as a key regulator of necroptosis, apoptosis, and inflammatory signaling in human and mammals. Here we report the identification of the RIPK3 homologue from black carp Mylopharyngodon piceus (bcRIPK3) and describe its role in regulating MAVS/IFN signaling. qPCR results demonstrated that bcRIPK3 was transcriptionally activated in response to poly (I:C) or LPS stimulation. Immunoblot assay and immunofluorescent staining assay showed that bcRIPK3 was a cytosolic protein with molecular weights of 47 kDa. Like its mammalian counterparts, bcRIPK3 exhibited a conserved function in inducing cell death. The reporter assay and plaque assay showed that overexpression of bcRIPK3 restricted bcMAVS-activated transcription of the interferon promoters of black carp and zebrafish, and suppressed bcMAVS-mediated antiviral activity. Notably, EPC cells co-expressing bcRIPK3, bcRIPK1 and bcMAVS presented much attenuated antiviral activity than the cells co-expressing bcRIPK3 and bcMAVS; and the subsequent co-IP assay identified the interaction between bcRIPK3 and bcRIPK1. Our findings collectively elucidate for the first time in teleost that black carp RIPK3 interacts with RIPK1 to inhibit MAVSmediated antiviral signaling.