Cai Mingsheng, Liao Zongmin, Chen Tao, Wang Ping, Zou Xingmei, Wang Yuanfang, Xu Zuo, Jiang Si, Huang Jinlu, Chen Daixiong, Peng Tao, Hong Gengde, Li Meili
Department of Pathogenic Biology and Immunology, Sino-French Hoffmann Institute, School of Basic Medical Science, Guangzhou Medical University, Guangzhou 511436, Guangdong, China.
Guangdong Haid Group Co., Ltd., Guangzhou 511400, Guandong, China.
Oncotarget. 2017 Jul 25;8(41):70006-70034. doi: 10.18632/oncotarget.19549. eCollection 2017 Sep 19.
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.
爱泼斯坦-巴尔病毒（EBV）是多种人类肿瘤的致病因素，但其某些编码蛋白尚未得到研究。作为功能鉴定的第一步，我们构建了一个包含大多数EBV编码蛋白的表达构建体文库，并明确了EBV在哺乳动物细胞中编码的81种蛋白的亚细胞定位图谱。将病毒开放阅读框与真核表达质粒中的增强型黄色荧光蛋白（EYFP）标签融合，然后在COS-7活细胞中表达，通过荧光显微镜观察蛋白定位。结果显示，所有蛋白中34.57%（28种蛋白）表现为全核或核内定位，39.51%（32种蛋白）表现为全细胞质或细胞质内定位，25.93%（21种蛋白）在细胞核和细胞质中均有发现。有趣的是，大多数包膜蛋白表现为全细胞质或膜定位，大多数衣壳蛋白在细胞核中显示富集或完全定位，这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相关。综上所述，EBV蛋白在活细胞中的亚细胞定位图谱可能为进一步阐明EBV编码基因在人类疾病尤其是相关肿瘤中的功能奠定基础。
