Negative regulation of interferon regulatory factor 6 (IRF6) in interferon and NF-κB signalling pathways of common carp (Cyprinus carpio L.).
Negative regulation of interferon regulatory factor 6 (IRF6) in interferon and NF-κB signalling pathways of common carp (Cyprinus carpio L.).
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DOI：
10.1186/s12917-022-03538-4
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发表时间：
2022-12-12
影响因子：
2.6
通讯作者：
Li, Hua
中科院分区：
文献类型：
--
作者：
Liang, Yaxin;Liu, Rongrong;Zhang, Jiahui;Chen, Yixin;Shan, Shijuan;Zhu, Yaoyao;Yang, Guiwen;Li, Hua
关键词：
干扰素(IFN)调节因子(IRFs)是一种转录因子，在调节I型干扰素及相关基因的表达中发挥重要作用。在哺乳动物中，IRF6 与 IFN 表达无关，而斑马鱼 IRF6 据报道是 IFN 表达的正调节因子，并且可以被 MyD88 和 TBK1 磷酸化。然而，IRF6在鲤鱼免疫反应和IFN转录中的作用尚不清楚。本研究采用RACE技术从鲤鱼中克隆了IRF6基因(CcIRF6)的cDNA，全长1905bp，编码471个氨基酸残基，具有DBD和IAD两个功能域。相似性分析表明，CcIRF6与其他脊椎动物的IRF相似性超过50%，与草鱼和斑马鱼的相似性最高，其中DBD结构域更为保守。系统发育分析表明，CcIRF6属于硬鱼纲，与草鱼亲缘关系最近。在健康鲤鱼中，CcIRF6在所有检测组织中均有表达，其中口腔上皮细胞中表达量最高，头肾中表达量最低。腹腔注射poly(I:C)或嗜水气单胞菌后，鲤鱼脾脏、头肾、前肠和后肠中CcIRF6的表达增加。此外，poly(I:C)、LPS、PGN和鞭毛蛋白在体外诱导鲤鱼外周血白细胞和头肾白细胞中CcIRF6的表达。在EPC细胞中，CcIRF6抑制一些IFN相关基因和促炎细胞因子的表达，双荧光素酶报告基因检测显示CcIRF6降低了IFN和NF-κB报告基因的活性。本研究提示CcIRF6参与鲤鱼的抗病毒和抗菌免疫反应，并负向调节IFN和NF-κB信号通路的表达，为鱼病发病机制的研究和预防提供理论依据。
Interferon (IFN) regulatory factors (IRFs) is a kind of transcription factors, which play an important role in regulating the expression of type I IFN and related genes. In mammals, IRF6 is not relevant with IFN expression, while zebrafish IRF6 was reported to be a positive regulator of IFN expression and could be phosphorylated by both MyD88 and TBK1. However, the role of IRF6 in the immune response and IFN transcription of common carp is unknown. In the present study, the cDNA of IRF6 gene (CcIRF6) was cloned from common carp using RACE technique, with a total length of 1905 bp, encoding 471 amino acid residues, which possesses two functional domains of DBD and IAD. Similarity analysis showed that CcIRF6 had more than 50% similarity with IRFs of other vertebrates, and had the highest similarity with grass carp and zebrafish, among which the DBD domain was much more conserved. The phylogenetic analysis showed that CcIRF6 is in the branch of Osteichthyes and has the closest relationship with grass carp. In healthy common carp, the CcIRF6 was expressed in all the examined tissues, with the highest level in the oral epithelium, and the lowest level in the head kidney. After intraperitoneal injection of poly(I:C) or Aeromonas hydrophila, the expression of CcIRF6 increased in spleen, head kidney, foregut and hindgut of common carp. Moreover, poly(I:C), LPS, PGN and flagellin induced the expression of CcIRF6 in peripheral leukocytes and head kidney leukocytes of common carp in vitro. In EPC cells, CcIRF6 inhibited the expression of some IFN-related genes and pro-inflammatory cytokines, and dual luciferase reporter assay showed that CcIRF6 reduced the activity of IFN and NF-κB reporter genes. The present study suggests that CcIRF6 is involved in the antiviral and antibacterial immune response of common carp, and negatively regulate the expression of IFN and NF-κB signalling pathways, which provides a theoretical basis for the study and prevention of fish disease pathogenesis.
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