Characterization of the subcellular localization of Epstein-Barr virus encoded proteins in live cells
Characterization of the subcellular localization of Epstein-Barr virus encoded proteins in live cells
复制标题
活细胞中 Epstein-Barr 病毒编码蛋白的亚细胞定位特征
DOI：
10.18632/oncotarget.19549
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发表时间：
2017-09-19
期刊：
影响因子：
--
通讯作者：
Li, Meili
中科院分区：
文献类型：
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作者：
Cai, Mingsheng;Liao, Zongmin;Li, Meili
EB病毒（Epstein-Barr virus，EBV）是多种人类肿瘤的致病因子，但其编码的蛋白质尚未被研究。作为功能鉴定的第一步，我们提出了大多数EBV编码蛋白的表达构建体文库的构建和哺乳动物细胞中EBV编码的81种蛋白的明确亚细胞定位图。将病毒开放阅读框与增强型黄色荧光蛋白（enhanced yellow fluorescent protein，EYFP）标签融合构建真核表达质粒，在COS-7细胞中表达，荧光显微镜下观察蛋白定位。结果表明，有34.57%（28个蛋白）的蛋白质定位于全核或亚核，39.51%（32个蛋白质）定位于胞质或亚胞质，25.93%（21个蛋白质）同时定位于胞核和胞质。有趣的是，大多数包膜蛋白呈现泛胞质或膜定位，而大多数衣壳蛋白在细胞核中显示富集或完全定位，表明特定蛋白的亚细胞定位与其在病毒复制过程中的作用有关。这些结果为进一步阐明EBV编码基因在人类疾病尤其是相关肿瘤中的功能奠定了基础。
Epstein-Barr virus (EBV) is the pathogenic factor of numerous human tumors, yet certain of its encoded proteins have not been studied. As a first step for functional identification, we presented the construction of a library of expression constructs for most of the EBV encoded proteins and an explicit subcellular localization map of 81 proteins encoded by EBV in mammalian cells. Viral open reading frames were fused with enhanced yellow fluorescent protein (EYFP) tag in eukaryotic expression plasmid then expressed in COS-7 live cells, and protein localizations were observed by fluorescence microscopy. As results, 34.57% (28 proteins) of all proteins showed pan-nuclear or subnuclear localization, 39.51% (32 proteins) exhibitted pan-cytoplasmic or subcytoplasmic localization, and 25.93% (21 proteins) were found in both the nucleus and cytoplasm. Interestingly, most envelope proteins presented pan-cytoplasmic or membranous localization, and most capsid proteins displayed enriched or complete localization in the nucleus, indicating that the subcellular localization of specific proteins are associated with their roles during viral replication. Taken together, the subcellular localization map of EBV proteins in live cells may lay the foundation for further illustrating the functions of EBV-encoded genes in human diseases especially in its relevant tumors.