摘要
肿瘤坏死因子受体相关因子2 (tumor necrosis factor receptor-associated factor 2, TRAF2)是一种重要的胞内信号接头蛋白,参与激活NF-κB和MAPK信号通路,在免疫防御、炎症反应和细胞凋亡等过程发挥关键作用。为了探索杂交鳢(Channa maculate♀×Channa argus♂) TRAF2功能及其对两种病原菌感染的应答机制,本研究克隆获得杂交鳢TRAF2 (命名为CmaTRAF2)基因的完整开放阅读框(ORF),长度为1 566 bp,预测编码521个氨基酸,与龟壳攀鲈(Anabas testudineus)、杜氏鰤(Seriola dumerili) TRAF2蛋白序列相似性最高,都达到91%。与其他TRAF2相似,CmaTRAF2具有一个RING结构域、一个锌指结构域、一个TRAF结构域。系统发育分析表明CmaTRAF2与其他硬骨鱼类TRAF2聚为一支,其中与龟壳攀鲈TRAF2的亲缘性关系最近。实时荧光定量PCR检测分析显示CmaTRAF2在健康杂交鳢所检测的9种组织中广泛分布,其中脾脏中表达量最高,头肾中的表达量次之,肝脏中的表达量最低。采用鰤诺卡氏菌(Nocardia seriolae)腹腔注射感染杂交鳢后,肝脏、脾脏和头肾中CmaTRAF2的表达在感染后不同时间点总体呈现为上调趋势,且分别在10 h、1 d和2 d时达到最大值(P<0.05)。舒伯特气单胞菌(Aeromonas schubertii)注射感染杂交鳢后,CmaTRAF2的表达变化相似,也主要呈现上调趋势,且分别在肝脏、脾脏和头肾中于12 h、5 d和2 d时达到峰值(P<0.05)。以上结果表明,CmaTRAF2参与了杂交鳢抵御鰤诺卡氏菌和舒伯特气单胞菌感染的免疫应答反应。本研究可为进一步探索鱼类TRAF2的免疫防御和信号转导机制提供基础资料。
Tumor necrosis factor receptor-associated factor 2(TRAF2) is an important intracellular signal adaptor protein, which participates in the activation of NF-κB and MAPK signaling pathways, and plays a crucial role in immune defense, inflammatory response and cell apoptosis. In order to explore the function of TRAF2 in hybrid snakehead(Channa maculate ♀ × Channa argus ♂) and its response to infection by two pathogens, the open reading frame(ORF) of the TRAF2(named: CmaTRAF2) was cloned from hybrid snakehead, which was 1 566 bp in full length and encoded 521 amino acids. The most similarity of CmaTRAF2 was found with TRAF2 proteins of Anabas testudineus, Seriola dumerili, reaching 91%. Similar to other TRAF2 proteins, CmaTRAF2 had one RING domain, one zinc finger domain, and one TRAF domain. In addition, phylogenetic analysis showed that CmaTRAF2 was clustered into one group with other teleost and had the closest relationship with TRAF2 of Anabas testudineus.The RT-qPCR results revealed that CmaTRAF2 was widely expressed in all nine tested tissues of healthy hybrid snakehead, with the highest expression in spleen, followed by head kidney, and the lowest in liver. The expressions of CmaTRAF2 in liver, spleen and head kidney were almost all up-regulated at different time points after infection with Nocardia seriolae through intraperitoneal injection, reaching maximum at 10 h, 1 d and 2 d, respectively(P<0.05). Similarly, the expressions of CmaTRAF2 in the liver, spleen, and head kidney of hybrid snakehead infected by Aeromonas schubertii were mainly up-regulated, reaching peak at 12 h, 5 d and 2 d, respectively(P<0.05). These results indicated that CmaTRAF2 might be involved in the immune response of hybrid snakehead and play a key role in the defense against the infection of two pathogens. This research presented basic data for further exploring the mechanism of immune defense and signal transduction of TRAF2 gene in fish.
作者
孔璐璐
赵飞
谭爱萍
刘付翠
邓玉婷
姜兰
Kong Lulu;Zhao Fei;Tan Aiping;Liu Fucui;Deng Yuting;Jiang Lan(Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province,Key Laboratory of Fishery Drug Development,of Ministry of Agriculture and Rural Affairs,Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou,510380;College of Fishery and Life Science,Shanghai Ocean University,Shanghai,201306)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2021年第1期101-108,共8页
Genomics and Applied Biology
基金
国家自然科学基金(31402338)
现代农业产业技术体系专项(CARS-46)
广东省自然科学基金(2015A030-310137)共同资助。
关键词
杂交鳢
肿瘤坏死因子受体相关因子2
应答分析
鰤诺卡氏菌
舒伯特气单胞菌
Hybrid snakehead(Channa maculate♀×Channa argus♂)
Tumor necrosis factor receptor-associated factor 2
Response analysis
Nocardia seriolae
Aeromonas schubertii
