PRMT6 inhibits K63-linked ubiquitination and promotes the degradation of IRF3 in the antiviral innate immunity of black carp Mylopharyngodon piceus
PRMT6 inhibits K63-linked ubiquitination and promotes the degradation of IRF3 in the antiviral innate immunity of black carp Mylopharyngodon piceus
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DOI：
10.1016/j.aquaculture.2022.738872
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发表时间：
2022-09
期刊：
影响因子：
4.5
通讯作者：
Can Yang;Shisi Yang;Yujia Miao;Juanjuan Shu;Yu Peng;Jun Yu Li;Hui-na Wu;Jun Zou;Hao Feng
中科院分区：
文献类型：
--
作者：
Can Yang;Shisi Yang;Yujia Miao;Juanjuan Shu;Yu Peng;Jun Yu Li;Hui-na Wu;Jun Zou;Hao Feng
蛋白质精氨酸甲基转移酶（PRMT 6）已被证明负调节干扰素（IFN）的生产在哺乳动物中，然而，硬骨鱼PRMT 6在IFN信号转导中的作用仍然在很大程度上未知。在我们之前的研究中，青鱼PRMT 6（bcPRMT 6）被鉴定为抑制bcTBK 1/bcIRF 3级联反应。在这项研究中，共表达的bcPRMT 6减弱了bcIRF 3诱导的宿主细胞中bcIFNa、bcIFNb、bcp 50和bcI 11 β的转录。因此，bcPRMT 6明显抑制了bcIRF 3介导的抗草鱼呼肠孤病毒（GCRV）和鲤鱼春季病毒血症病毒（SVCV）的活性。bcPRMT 6基因的敲除增强了宿主细胞的抗病毒能力，并提高了bcIFNa、bcViperin和bcI 11 β的转录水平。当与bcPRMT 6在哺乳动物或鱼类细胞中共表达时，bcIRF 3的蛋白水平降低，这意味着bcPRMT 6触发bcIRF 3的降解。有趣的是，这种降低的bcIRF 3蛋白表达被氯喹而不是MG 132所拯救，表明这种分子的溶酶体依赖性降解。随后的免疫沉淀和免疫印迹试验表明，bcPRMT 6明显减弱了bcIRF 3的K63连接的泛素化，而不是K48连接的泛素化。免疫荧光染色结果表明，bcPRMT 6可抑制bcIRF 3的核转位。综上所述，我们的数据得出结论，bcPRMT 6通过溶酶体途径促进bcIRF 3的降解，并抑制bcIRF 3的K63连接的泛素化，从而抑制bcIRF 3介导的抗病毒信号传导。
Protein arginine methyltransferases (PRMT6) has been shown to negatively regulate interferon (IFN) production in mammals, however, the role of teleost PRMT6 in IFN signaling remains largely unknown. In our previous study, black carp (Mylopharyngodon piceus) PRMT6 (bcPRMT6) has been identified to inhibit bcTBK1/bcIRF3 cascade. In this study, bcIRF3-induced transcription ofbcIFNa, bcIFNb, bcp50 and bcIl1βin host cells was attenuated by co-expressed bcPRMT6. Accordingly, bcIRF3-mediated antiviral activity against grass carp reovirus (GCRV) and spring viremia of carp virus (SVCV) was obviously dampened by bcPRMT6. Knockdown of bcPRMT6 enhanced the antiviral ability of host cells and improved the transcription ofbcIFNa, bcViperin and bcIl1β. When co-expressed with bcPRMT6 in either mammalian or fish cells, the protein level of bcIRF3 was decreased, which implied the triggered degradation of bcIRF3 by bcPRMT6. And interestingly, this decreased protein expression of bcIRF3 was rescued by chloroquine but not MG132, indicating the lysosome-dependent degradation of this molecule. The subsequent immunoprecipitation and immunoblot assay demonstrated that K63-linked ubiquitination but not K48-linked of bcIRF3 was obviously attenuated by bcPRMT6. And the immunofluorescent staining data suggested that the nuclear translocation of bcIRF3 was inhibited by bcPRMT6. Taken together, our data concludes that bcPRMT6 promotes the degradation of bcIRF3 through lysosome pathway and inhibits K63-linked ubiquitination of bcIRF3, leading to the dampened antiviral signaling mediated by bcIRF3.