Section of Virology, Imperial College Faculty of Medicine, St. Mary's Campus, Norfolk Place, London W2 1PG, United Kingdom.
J Virol. 2011 Apr;85(7):3535-45. doi: 10.1128/JVI.02086-10. Epub 2011 Jan 19.
Novel Epstein-Barr Virus (EBV) strains with deletion of either EBER1 or EBER2 and corresponding revertant viruses were constructed and used to infect B lymphocytes to make lymphoblastoid cell lines (LCLs). The LCLs were used in microarray expression profiling to identify genes whose expression correlates with the presence of EBER1 or EBER2. Functions of regulated genes identified in the microarray analysis include membrane signaling, regulation of apoptosis, and the interferon/antiviral response. Although most emphasis has previously been given to EBER1 because it is more abundant than EBER2, the differences in cell gene expression were greater with EBER2 deletion. In this system, deletion of EBER1 or EBER2 had little effect on the EBV transformation frequency of primary B cells or the growth of the resulting LCLs. Using the recombinant viruses and novel EBER expression vectors, the nuclear redistribution of rpL22 protein by EBER1 in 293 cells was confirmed, but in LCLs almost all of the cells had a predominantly cytoplasmic expression of this ribosomal protein, which was not detectably changed by EBER1. The changes in LCL gene expression identified here will provide a basis for identifying the mechanisms of action of EBER RNAs.
新型 Epstein-Barr 病毒 (EBV) 株缺失 EBER1 或 EBER2,并构建了相应的回复病毒,用于感染 B 淋巴细胞制作淋巴母细胞系 (LCL)。利用这些 LCL 进行微阵列表达谱分析,以鉴定与 EBER1 或 EBER2 存在相关的表达基因。在微阵列分析中鉴定的调控基因的功能包括膜信号转导、细胞凋亡调控和干扰素/抗病毒反应。尽管以前大多数研究都集中在 EBER1 上,因为它比 EBER2 更丰富,但 EBER2 缺失导致的细胞基因表达差异更大。在这个系统中,EBER1 或 EBER2 的缺失对原代 B 细胞的 EBV 转化频率或产生的 LCL 的生长几乎没有影响。使用重组病毒和新型 EBER 表达载体,在 293 细胞中证实了 EBER1 对 rpL22 蛋白的核重分布,但在 LCL 中,几乎所有细胞的这种核糖体蛋白都主要表达在细胞质中,EBER1 对其表达没有明显改变。这里鉴定的 LCL 基因表达变化将为鉴定 EBER RNA 的作用机制提供基础。