Mx1 of black carp functions importantly in the antiviral innate immune response
Mx1 of black carp functions importantly in the antiviral innate immune response
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青鱼的MX1在抗病毒先天免疫反应中发挥重要作用
DOI：
10.1016/j.fsi.2016.10.002
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发表时间：
2016
影响因子：
4.7
通讯作者：
Tang Luosheng
中科院分区：
文献类型：
--
作者：
Xiao Jun;Yan Jun;Chen Hui;Li Jun;Tian Yu;Feng Hao;Tang Luosheng
粘病毒抗性(MX)是脊椎动物对微生物病原体的先天免疫反应中的一种重要的抗病毒蛋白。在本研究中，我们克隆并鉴定了青鱼Mx1基因。青鱼Mx1(BcMx1)基因全长由2781个核苷酸组成，编码的bcMx1蛋白由631个氨基酸组成。BcMx1在N-末端含有一个GTP酶结构域，在中间有一个“中央相互作用结构域”，在C-末端有一个GTP酶效应结构域。BcMx1基因在心、肝、脾、肾、肠、肌肉、皮肤和鳃等组织中均有转录，感染草鱼呼肠孤病毒(GCRV)或病毒血症(SVCV)后，除鳃组织外，其余组织中bcMx1基因表达水平均升高。Mylphyngodon piceusfin(MPF)细胞的定量PCR分析表明，GCRV或SVCV感染MPF细胞后，bcMx1mRNA水平在不同的感染复数(MOI)下均升高。Western blotting表明，bcMx1的相对分子质量为∼75kDa，免疫荧光染色结果表明，bcMx1是一种胞浆蛋白。表达bcMx1基因的EPC细胞对SVCV和GCRV的抗病毒活性增强。我们的所有数据表明，bcMx1是青鱼先天免疫反应中的一种抗病毒蛋白。
Mx (myxovirus resistance) is an important antiviral protein in the innate immune responses of vertebrates to microbial pathogens. In this study, we cloned and characterized Mx1 of black carp (Mylopharyngodon piceus). The full-length cDNA of black carp Mx1 (bcMx1) consists of 2781 nucleotides and the predicted bcMx1 protein contains 631 amino acids. bcMx1 contains a GTPase domain at the N-terminnus, a “central interactive domain” in the middle and a GTPase effector domain at the C-terminus. bcMx1 mRNA was constitutively transcribed in all tissues tested, including the heart, liver, spleen, kidney, intestine, muscle, skin and gill; and bcMx1 mRNA levels increased in all but the gill after grass carp reovirus (GCRV) or viraemia of carp virus (SVCV) infection. Quantitative PCR analysis ofMylopharyngodon piceusfin (MPF) cells indicated that bcMx1 mRNA levels increased after GCRV or SVCV infection at different multiplicities of infection (MOI). Western blotting demonstrated that the molecular weight of bcMx1 is ∼75 kDa and immunofluorescent staining data of both HeLa cells and EPC cells showed that bcMx1 is a cytosolic protein. EPC cells transfected with plasmid expressing bcMx1 showed increased antiviral activity against SVCV and GCRV. All our data suggest that bcMx1 is an antiviral protein in the innate immune response of the black carp.