Channing Laboratory, Brigham and Women's Hospital and Harvard Medical School, 181 Longwood Avenue, Boston, MA 02115, USA.
J Virol. 2011 Jul;85(13):6764-73. doi: 10.1128/JVI.00422-11. Epub 2011 May 4.
Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) transforms rodent fibroblasts and is expressed in most EBV-associated malignancies. LMP1 (transformation effector site 2 [TES2]/C-terminal activation region 2 [CTAR2]) activates NF-κB, p38, Jun N-terminal protein kinase (JNK), extracellular signal-regulated kinase (ERK), and interferon regulatory factor 7 (IRF7) pathways. We have investigated LMP1 TES2 genome-wide RNA effects at 4 time points after LMP1 TES2 expression in HEK-293 cells. By using a false discovery rate (FDR) of <0.001 after correction for multiple hypotheses, LMP1 TES2 caused >2-fold changes in 1,916 mRNAs; 1,479 RNAs were upregulated and 437 were downregulated. In contrast to tumor necrosis factor alpha (TNF-α) stimulation, which transiently upregulates many target genes, LMP1 TES2 maintained most RNA effects through the time course, despite robust and sustained induction of negative feedback regulators, such as IκBα and A20. LMP1 TES2-regulated RNAs encode many NF-κB signaling proteins and secondary interacting proteins. Consequently, many LMP1 TES2-regulated RNAs encode proteins that form an extensive interactome. Gene set enrichment analyses found LMP1 TES2-upregulated genes to be significantly enriched for pathways in cancer, B- and T-cell receptor signaling, and Toll-like receptor signaling. Surprisingly, LMP1 TES2 and IκBα superrepressor coexpression decreased LMP1 TES2 RNA effects to only 5 RNAs, with FDRs of <0.001-fold and >2-fold changes. Thus, canonical NF-κB activation is critical for almost all LMP1 TES2 RNA effects in HEK-293 cells and a more significant therapeutic target than previously appreciated.
EB 病毒（EBV）潜伏膜蛋白 1（LMP1）转化啮齿动物成纤维细胞，并在大多数 EBV 相关恶性肿瘤中表达。LMP1（转化效应元件 2 [TES2]/C 端激活区 2 [CTAR2]）激活 NF-κB、p38、Jun N 端蛋白激酶（JNK）、细胞外信号调节激酶（ERK）和干扰素调节因子 7（IRF7）途径。我们在 HEK-293 细胞中表达 LMP1 TES2 后 4 个时间点，对 LMP1 TES2 基因组范围的 RNA 效应进行了研究。通过对多个假设进行修正后，FDR<0.001，LMP1 TES2 导致 1916 个 mRNA 的倍数变化>2 倍；1479 个 RNA 上调，437 个 RNA 下调。与肿瘤坏死因子-α（TNF-α）刺激短暂地上调许多靶基因不同，尽管负反馈调节剂（如 IκBα 和 A20）的诱导强烈且持续，但 LMP1 TES2 通过时间过程维持了大多数 RNA 效应。LMP1 TES2 调节的 RNA 编码许多 NF-κB 信号蛋白和二级相互作用蛋白。因此，许多 LMP1 TES2 调节的 RNA 编码形成广泛相互作用组的蛋白质。基因集富集分析发现，LMP1 TES2 上调的基因在癌症、B 和 T 细胞受体信号以及 Toll 样受体信号通路中显著富集。令人惊讶的是，LMP1 TES2 和 IκBα 超级阻遏物共表达将 LMP1 TES2 RNA 效应降低至仅 5 个 RNA，FDR<0.001 倍和>2 倍变化。因此，在 HEK-293 细胞中，经典 NF-κB 激活对于 LMP1 TES2 的几乎所有 RNA 效应都是至关重要的，并且是比以前认识到的更重要的治疗靶标。
