Zhou Wei, Zhou Jujun, Lv Ying, Qu Yixiao, Chi Mengdie, Li Jun, Feng Hao
Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education of China, College of Life Science, Hunan Normal University, Changsha, 410081, China.
Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education of China, College of Life Science, Hunan Normal University, Changsha, 410081, China; State Key Laboratory of Freshwater Ecology and Biotechnology, Wuhan, 430072, China.
Fish Shellfish Immunol. 2015 Apr;43(2):460-8. doi: 10.1016/j.fsi.2015.01.016. Epub 2015 Feb 3.
MAVS (mitochondria antiviral signaling protein) plays an important role in the host cellular innate immune response against microbial pathogens. In this study, MAVS has been cloned and characterized from black carp (Mylopharyngodon piceus). The full-length cDNA of black carp MAVS (bcMAVS) consists of 2352 nucleotides and the predicted bcMAVS protein contains 579 amino acids. Structural analysis showed that bcMAVS is composed of functional domains including an N-terminal CARD, a central proline-rich domain, a putative TRAF2-binding motif and a C-terminal TM domain, which is similar to mammalian MAVS. bcMAVS is constitutively transcribed in all the selected tissues including gill, kidney, heart, intestine, liver, muscle, skin and spleen; bcMAVS mRNA level in intestine, liver, muscle increased but decreased in spleen right after GCRV or SVCV infection. Multiple bands of bcMAVS were detected in western blot when it was expressed in tissue culture, which is similar to mammalian MAVS. Immunofluorescence assay determined that bcMAVS is a mitochondria protein and luciferase reporter assay demonstrated that bcMAVS could induce zebrafish IFN and EPC IFN expression in tissue culture. Data generated in this manuscript has built a solid foundation for further elucidating the function of bcMAVS in the innate immune system of black carp.
线粒体抗病毒信号蛋白（MAVS）在宿主细胞针对微生物病原体的固有免疫反应中发挥着重要作用。在本研究中，已从草鱼（Mylopharyngodon piceus）中克隆并鉴定了MAVS。草鱼MAVS（bcMAVS）的全长cDNA由2352个核苷酸组成，预测的bcMAVS蛋白包含579个氨基酸。结构分析表明，bcMAVS由包括N端CARD、中央富含脯氨酸结构域、一个假定的TRAF2结合基序和一个C端跨膜结构域的功能结构域组成，这与哺乳动物的MAVS相似。bcMAVS在所有选定的组织中组成性转录，包括鳃、肾、心脏、肠道、肝脏、肌肉、皮肤和脾脏；在感染GCRV或SVCV后，肠道、肝脏、肌肉中的bcMAVS mRNA水平升高，但脾脏中的bcMAVS mRNA水平下降。当bcMAVS在组织培养中表达时，在蛋白质免疫印迹中检测到多条带，这与哺乳动物的MAVS相似。免疫荧光分析确定bcMAVS是一种线粒体蛋白，荧光素酶报告基因分析表明bcMAVS可以在组织培养中诱导斑马鱼IFN和EPC IFN表达。本研究中产生的数据为进一步阐明bcMAVS在草鱼固有免疫系统中的功能奠定了坚实的基础。
