摘要
在脊椎动物中viperin已被证明是一种能抵御大多数DNA和RNA病毒的抗病毒蛋白质。本文克隆及鉴定了青鱼viperin。青鱼viperin(bc Viperin)的c DNA由1 828个核苷酸组成,编码360个氨基酸。预测的bc Viperin蛋白包含N端低保守的两性α-螺旋结构域、中间自由基S-腺苷甲硫氨酸(S-adenosylmethionine,SAM)结构域及高保守C端结构域。bc Viperin m RNA在青鱼心脏、肝脏、脾脏、肾脏、肠、肌肉、皮肤和鳃等组织中均有表达。在草鱼呼肠孤病毒(grass carp reovirus,GCRV)和鲤春病毒血症病毒(spring viremia of carp virus,SVCV)感染后,bc Viperin m RNA在上述组织中的表达水平提高。在青鱼尾鳍(Mylopharyngodon piceus fin,MPF)细胞中,bc Viperin m RNA表达水平同样会在GCRV和SVCV感染后发生上升。蛋白质免疫印迹表明bc Viperin蛋白的相对分子质量约为42 k D;He La和EPC细胞中的免疫荧光染色实验表明bc Viperin是细胞质蛋白。编码bc Viperin的质粒在EPC细胞中过表达后能够显著提高细胞抵御SVCV和GCRV的抗病毒能力。以上研究表明bc Viperin是青鱼天然免疫过程中重要的抗病毒蛋白质。
Viperin has been shown to be an antiviral protein against a wide range of DNA and RNA viruses in vertebrates. Herein, the viperin of black carp Mylopharyngodon piceus has been cloned and characterized. The full-length c DNA of black carp viperin（bc Viperin） consists of 1 828 nucleotides and the predicted bc Viperin protein contains 360 amino acids, which is composed of a less conserved N-terminal amphipathic α-helix, a central radical S-adenosylmethionine（SAM） and a highly conserved C-terminal region. bc Viperin m RNA has been constitutively detected in all the selected tissues of black carp, including the heart, liver, spleen, kid-ney, intestine, muscle, skin and gills. The m RNA level of bc Viperin increased in all these tissues after both GCRV and SVCV infections. q-PCR data demonstrated that bc Viperin m RNA level in Mylopharyngodon piceus fin（MPF） cells also increased in response to both GCRV and SVCV infections. Western-blot demonstrated that the relative molecular mass of bc Viperin is around 42 k D; and the immunofluorescence staining of both He La and EPC cells showed that bc Viperin is a cytosolic protein. In addition, EPC cells transfected with plasmid expressing bc Viperin presented obviously improved antiviral ability against both SVCV and GCRV. Taken together, these findings support the conclusion that bc Viperin is an antiviral protein functioning in the innate immunity of black carp.
作者
田彧
陈慧
周旻昱
李俊
肖俊
冯浩
TIAN Yu;CHEN Hui;ZHOU nin-yu;LI Jun;XIAO Jun;FENG Hao(State Key Laboratory of Developmental Biology of Freshwater Fish,College of Life Science,Hunan Normal University,Changsha 410081,Hunan,China;The Second Xiangya Hospital,Central South University,Changsha 410011,Hunan,China;College of Bioscience and Biatechnology,Hunan Agricultural University,Changsha 410128,Hunan,China)
出处
《生命科学研究》
CAS
CSCD
2018年第4期259-270,共12页
Life Science Research
基金
The National Natural Science Foundation of China(81471963)
Hunan Provincial Innovation Foundation for Postgraduate(CX2017B185)~~
