Identification, characterisation and preliminary functional analysis of IRAK-M in grass carp (Ctenopharyngodon idella)
Identification, characterisation and preliminary functional analysis of IRAK-M in grass carp (Ctenopharyngodon idella)
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草鱼IRAK-M的鉴定、表征及初步功能分析
DOI：
10.1016/j.fsi.2018.09.080
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发表时间：
2019
影响因子：
4.7
通讯作者：
Wang Yaping
中科院分区：
文献类型：
--
作者：
Chu Pengfei;He Libo;Zhu Denghui;Chen Liangming;Huang Rong;Liao Lanjie;Li Yongming;Zhu Zuoyan;Wang Yaping
白细胞介素-1受体相关激酶（IRAK）家族成员在髓样分化初级反应88 （MyD88）依赖toll样受体（TLR）信号传导中发挥重要作用，这是脊椎动物重要的先天免疫途径。本研究对草鱼IRAK家族基因IRAK- m（也称为dirak3）进行了克隆和鉴定。irak - m主要富集于脾脏，在草鱼呼肠孤病毒（GCRV）感染后表达显著改变。亚细胞定位结果显示，IRAK-M蛋白在整个细胞内分布均匀，并与MyD88在转染细胞的细胞质内共定位。此外，通过双分子荧光互补（BiFC）系统证实了IRAK-M与MyD88之间的相互作用。此外，缺乏IRAK-M的小干扰RNA （siRNA）的idellakidney细胞系（CIK）上调多肌苷：多胞酸（poly(I:C)）诱导的炎症细胞因子的产生，包括白细胞介素8 （IL-8）、IL-6和肿瘤坏死因子α(TNF-α)，这表明IRAK-M是炎症细胞因子的负调节因子。综上所述，我们的研究结果表明，irak - mgene在固有免疫调节中起着重要作用，并为理解IRAK-M在硬骨鱼中的功能特征提供了新的见解。
Interleukin-1 receptor-associated kinase (IRAK) family members play important roles in myeloid differentiation primary response 88 (MyD88)-dependent toll-like receptor (TLR) signaling, the crucial innate immune pathway in vertebrates. In the present study, the IRAK family geneIRAK-M(also calledIRAK3) from grass carp (Ctenopharyngodon idella) was cloned and characterised.IRAK-Mwas mainly enriched in the spleen, and the significantly altered expression was observed after grass carp reovirus (GCRV) infection. Subcellular localisation showed that IRAK-M protein distributed uniformly in the entire cell and co-localised with MyD88 in the cytoplasm of transfected cells. Additionally, the interaction between IRAK-M and MyD88 was confirmed by bimolecular fluorescence complementation (BiFC) system. Moreover, deficient of IRAK-M inC. idellakidney cell line (CIK) with small interference RNA (siRNA) upregulated polyinosinic:polycytidylic acid (poly(I:C))-induced inflammatory cytokines production, including interleukin 8 (IL-8),IL-6, and tumour necrosis factorα(TNF-α), which reveals that IRAK-M functions as a negative regulator of inflammatory cytokines. Taken together, our results demonstrate thatIRAK-Mgene plays an important role in innate immune regulation and provide new insights into understanding the functional characteristics of the IRAK-M in teleosts.