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小麦黄花叶病毒编码VPg蛋白N端结构是VPg与核仁蛋白Fibrillarin互作区域
引用本文:边靓,向荣,孙丽英,陈剑平.小麦黄花叶病毒编码VPg蛋白N端结构是VPg与核仁蛋白Fibrillarin互作区域[J].浙江农业学报,2014,26(1):0.
作者姓名:边靓  向荣  孙丽英  陈剑平
作者单位:1.浙江师范大学 化学与生命科学学院,浙江 金华321004;2浙江省农业科学院 病毒学与生物技术研究所/浙江省植物有害生物防控国家重点实验室培育基地/农业部植物保护与生物技术重点开放实验室/浙江省植物病毒重点开放实验室,浙江 杭州310021
摘    要:小麦黄花叶病毒(Wheat yellow mosaic virus,WYMV)属于马铃薯Y病毒科大麦黄花叶病毒属成员,其基因组是由两条正义单链RNA组成,共编码10个蛋白。其中,VPg(Viral protein genome\|linked)作为病毒末端结合蛋白,与病毒基因组RNA 5’端共价连接。利用软件分析WYMV VPg蛋白氨基酸序列发现其N端具有核定位信号(NLS)序列,C端具有核输出信号(NES)序列。通过激光共聚焦显微镜观察VPg与GFP融合蛋白在烟草细胞中的表达情况发现,GFP\|VPg主要定位于细胞核中。利用eYFP(n)标记核仁结构蛋白Fibrillarin与VPg进行双分子荧光互补实验发现,VPg与Fibrillarin互作且定位于细胞核中。根据VPg结构特点构建一系列缺失突变体与Fibrillarin的互作实验验证了它们之间的互作区域发生在N端NLS区域。

关 键 词:小麦黄花叶病毒  VPg蛋白  细胞核定位  Fibrillarin蛋白  蛋白质互作  

N\|terminal domain of VPg encoded by Wheat yellow mosaic virus is involved in the interaction with the nucleolar protein Fibrillarin
BIAN Jing,XIANG Rong,SUN Li\|ying,CHEN Jian\|ping.N\|terminal domain of VPg encoded by Wheat yellow mosaic virus is involved in the interaction with the nucleolar protein Fibrillarin[J].Acta Agriculturae Zhejiangensis,2014,26(1):0.
Authors:BIAN Jing  XIANG Rong  SUN Li\|ying  CHEN Jian\|ping
Institution:1College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China;2State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control/Ministry of Agriculture Key Laboratory of Biotechnology in Plant Protection/Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China)
Abstract:Wheat yellow mosaic virus (WYMV) is a member of the genus Bymovirus in the family Potyviridae. WYMV has two single\|stranded positive sense RNA genomes which produce 10 viral proteins. Among those proteins, viral genome\|linked protein (VPg), which binds to the 5′ end of the viral RNA genome, is predicted to contain a nuclear localization signal (NLS) and nuclear export signal (NES) in the N\| and C\|terminal portions, respectively. WYMV(Wheat yellow mosaic virus) VPg fused to C\|terminus of enhanced green florescent protein (eGFP\|VPg) exclusively accumulated in the nucleus of epidermal cells of Nicotiana benthamiana plants. Fibrillarin, a nucleolus resident protein, was introduced into eYFP(n) plasmid. To investigate the regions of the VPg domain binding fibrillarin, a series of VPg mutant and fibrillarin performed BiFC assay. The result display VPg localized and interact with fibrillarin in the nucleus. NLS but not NES is required for interact with fibrillarin.
Keywords:Wheat yellow mosaic virus  VPg protein  nuclear localization  fibrillarin  protein interaction  
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