摘要
蓝舌病病毒(bluetongue virus,BTV)编码4种非结构蛋白(NS1~NS4),其中NS4是新发现的非结构蛋白。为深入探索该新型非结构蛋白的功能,本研究首先采用生物信息学分析其序列结构特征及可能的功能域,结果发现NS4蛋白的N端α-螺旋结构域中具有富含碱性氨基酸的核定位信号(NLS),而C端α-螺旋结构域中具有亮氨酸拉链结构的核输出信号(NES)。为进一步证实NS4的亚细胞定位,本试验构建了表达NS4全长及其缺失突变体的重组质粒,结果显示缺失NLS的NS4重组蛋白弥散在胞浆,而缺失NES后NS4重组蛋白只定位于细胞核,表明NS4蛋白亚细胞定位具有核—胞浆穿梭过程。最后结合3D同源建模发现NS4与CCAAT增强子结合蛋白(C/EBPβ)等具有较高的同源性,据此推测NS4可能为转录调控蛋白,与DNA或其他转录因子存在相互作用。总之,本研究为NS4蛋白的功能研究提供了更多新线索和新思路,填补了BTV基础研究的空白。
Bluetongue virus(BTV)could code four non-structural proteins(NS1to NS4),in which NS4 was a novel one.To explore the function of NS4 protein of BTV,bioinformatics was firstly used to analyze the structure characteristics and possible function motifs of NS4 protein,the results showed that a basic area at the N-terminalα-coil contained a nuclear localization signal(NLS)and a leucine zipper area at the C-terminalα-coil contained a nuclear export signal(NES).To further confirm the subcellular localization of NS4 protein,the full-length and mutant NS4 lacking NLS or NES were expressed fusing with enhanced green fluorescence protein(EGFP).The results showed that NS4 recombinant protein lacking NLS was dispersed in the cytoplasm,while NS4 was located in the nucleus when NES was missing,which suggested that NS4 protein might have nuclear-cytoplasm shuttling process.Additionally,3Dhomology modeling showed that NS4 protein was similar to transcription factors CCAAT enhancer binding protein beta(C/EBPβ)and so on.So we speculated that NS4 protein was a transcription regulatory protein interacting with DNA or other transcription factors.In conclusion,this study provided new clues and ideas for function research of NS4 protein,and filled the gap of basic research areas of BTV.
作者
吕爽
杨涛
孙恩成
徐青元
张继凯
吴东来
LV Shuang;YANG Tao;SUN En-cheng;XU Qing-yuan;ZHANG Ji-kai;WU Dong-ai(Key Laboratory of Veterinary Public Health,Ministry of Agriculture,State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agriculture Sciences,Harbin 150001,China)
出处
《中国畜牧兽医》
CAS
北大核心
2016年第1期1-7,共7页
China Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金(31302065)
国家科技支撑计划(2013BAD12B05)
公益性行业(农业)科研专项资助(201203056)
中央级公益性科研院所基本科研业务费(0302015008)
关键词
蓝舌病病毒
NS4蛋白
亚细胞定位
同源建模
bluetongue virus(BTV)
non-structural protein 4(NS4)
subcellular localization
homology modeling
