Biological characterization, expression, and functional analysis of tumor necrosis factor receptor-associated factor 6 in Nile tilapia (Oreochromis niloticus)
Biological characterization, expression, and functional analysis of tumor necrosis factor receptor-associated factor 6 in Nile tilapia (Oreochromis niloticus)
复制标题
尼罗罗非鱼 (Oreochromis niloticus) 肿瘤坏死因子受体相关因子 6 的生物学特性、表达和功能分析
DOI：
10.1016/j.fsi.2018.06.036
复制
发表时间：
2018
影响因子：
4.7
通讯作者：
Jian JiChang
中科院分区：
文献类型：
--
作者：
Wang Zhiwen;Huang Yu;Li Yuan;Wang Bei;Lu Yishan;Xia Liqun;Tang Jufen;Jian JiChang
尼罗罗非鱼（Oreochromis niloticus）是我国重要的经济鱼类，无乳链球菌感染一直困扰着我国。肿瘤坏死因子受体相关因子6（tumor necrosis factor receptor associated factor 6，TRAF 6）是白细胞介素1受体/Toll样受体（interleukin-1 receptor/Toll-like receptor，IL-1/TLR）超家族中的一个重要的衔接分子，可触发参与先天免疫的下游信号级联反应。本研究从O. niloticus（命名为On-TRAF 6），其开放阅读框为1716 bp，编码571个氨基酸的多肽。On-TRAF 6的氨基酸序列包含TRAF蛋白的特征性基序，包括一个RING型锌指结构域、两个TRAF型锌指结构域和一个MATH（meprin and TRAF homology）结构域。多序列比对显示On-TRAF 6与其他鱼类的同源性较高（64-98%）。On-TRAF 6 mRNA在健康罗非鱼各组织中均有表达，其中脾脏中的表达量最高。此外，通过qPCR检测发现On-TRAF 6参与了尼罗罗非鱼在无乳链球菌和聚肌胞刺激下的免疫应答。亚细胞定位结果显示On-TRAF 6主要分布于细胞质中，过表达On-TRAF 6可显著激活NF-кB通路。这些结果表明On-TRAF 6可能在尼罗罗非鱼对胞内细菌的免疫应答中起重要作用。
Nile tilapia (Oreochromis niloticus) is a pivotal economic fish that has been plagued byStreptococcus agalactiaeinfections for many years. Tumor necrosis factor receptor-associated factor 6 (TRAF6) is a crucial adaptor molecule of the interleukin-1 receptor/Toll-like receptor (IL-1/TLR) superfamily, which can trigger downstream signaling cascades involved in innate immunity. In this study, the full-length cDNA ofTRAF6was cloned fromO. niloticus(namedOn-TRAF6), which has an open reading frame of 1716 bp, and encode a polypeptide of 571 amino acids. The predicted amino acid sequence of On-TRAF6 contained the characteristic motifs of TRAF proteins, including a Zinc finger of RING-type, two Zinc fingers of TRAF-type, and a MATH (meprin and TRAF homology) domain. Multiple sequence alignment revealed that On-TRAF6 shares a relatively high level of identity with those of other fishes (64–98%). In healthy tilapia, mRNA expression ofOn-TRAF6could be detected in all the examined tissues and the highest expression occurred in the spleen. Moreover, we found thatOn-TRAF6was involved immune response of Nile tilapia following the stimulation withStreptococcus agalactiaeand polyinosinic: polycytidylic acid (Poly I:C) when determined by using qPCR. The result of subcellular localization showed that On-TRAF6 distributed in the cytoplasm, and over-expression of On-TRAF6 could strongly activated NF-кB pathway in HEK293T cells. These findings indicated that On-TRAF6 may play an important role in the immune response to intracellular bacteria in Nile tilapia.