青岛文昌鱼AHR和ARNT的鉴定和表达分析及相互作用研究  

Identification,Expression and Interaction Analysis of AHR and ARNT in Branchiostoma japonicum

作　　者：刘颖超 程潇 宋丽丽 张宇 Liu Yingchao;Cheng Xiao;Song Lili;Zhang Yu(College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China)

机构地区：[1]中国海洋大学海洋生命学院,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2022年第6期79-91,共13页Periodical of Ocean University of China

基　　金：国家自然科学基金项目(31501856);山东省自然科学基金项目(ZR2015CQ004);青岛市应用基础研究计划项目(18-2-2-29-jch)资助。

摘　　要：为了探究文昌鱼中芳香烃受体(Aryl hydrocarbon receptor,AHR)和芳香烃受体核转运蛋白(Aryl hydrocarbon receptor nuclear translocator,ARNT)是否参与化学防御以及是否存在类似脊椎动物的AHR-CYP信号通路,本研究通过基因克隆、系统进化分析、表达分析和蛋白互作分析等方法进行探究。本研究从青岛文昌鱼(Branchiostoma japonicum)中鉴定出Bjahr与Bjarnt基因,系统进化分析显示Bjahr与Bjarnt较为原始。组织表达分析显示Bjahr与Bjarnt在检测的7种组织中均有表达,且均在卵巢、精巢和脊索中表达量较高。类二噁英PCB126胁迫文昌鱼后,Bjahr、Bjarnt及生物转化酶基因cyp1、cyp3、gst和sult1a在肝盲囊中的表达都有显著升高,说明这些基因参与文昌鱼的化学防御。此外,研究中还获得了文昌鱼AHR功能域部分和ARNT的体外重组蛋白。ELISA结合实验显示,文昌鱼AHR功能域能够与ARNT结合,且脊椎动物AHR的配体PCB126能够促进这两个蛋白的结合。这些结果说明,文昌鱼AHR和ARNT可能具有脊椎动物同源蛋白类似的化学防御功能,即文昌鱼中可能存在AHR-CYP信号通路。In order to explore whether Aryl hydrocarbon receptor(AHR)and Aryl hydrocarbon receptor nuclear translocation(ARNT)in amphioxus are involved in chemical defense and whether there is a vertebrate like AHR-CYP signal pathway,this study was carried out by gene cloning,phylogenetic analysis,expression analysis and protein interaction analysis.In this study,Bjahr and Bjarnt genes are identified from amphioxus(Branchiostoma japonicum).Phylogenetic analysis showed that Bjahr and Bjarnt represented ancestor type.The expression analysis showed that both Bjahr and Bjarnt were expressed in all examined tissues and predominantly expressed in the ovary,testis and notochord.The expressions of Bjahr,Bjarnt and biotransformation enzyme genes cyp1,cyp3,gst and sult1a were significantly increased in the hepatic cecum after exposed to PCB126,suggesting that these genes are involved in the chemical defense.In addition,we obtained the recombinant proteins of AHR domain and ARNT in vitro by prokaryotic expression.ELISA binding analysis showed that amphioxus AHR domain could bind to ARNT.PCB126,the ligand of vertebrates AHR,could promote the affinity of the two proteins.These results suggested that the function of amphioxus AHR and ARNT are probably similar to vertebrate homologues in response to xenobiotic chemical,and AHR-CYP pathway may exist in amphioxus.

关 键 词：青岛文昌鱼 芳香烃受体 芳香烃受体核转运蛋白 AHR-CYP信号通路 化学防御系统 

分 类 号：Q173[生物学—水生生物学]