Epstein-Barr virus (EBV) transforms B cells to continuously proliferating lymphoblastoid cell lines (LCLs), which represent an experimental model for EBV-associated cancers. EBV nuclear antigens (EBNAs) and LMP1 are EBV transcriptional regulators that are essential for LCL establishment, proliferation, and survival. Starting with the 3D genome organization map of LCL, we constructed a comprehensive EBV regulome encompassing 1,992 viral/cellular genes and enhancers. Approximately 30% of genes essential for LCL growth were linked to EBV enhancers. Deleting EBNA2 sites significantly reduced their target gene expression. Additional EBV super-enhancer (ESE) targets included MCL1, IRF4, and EBF. MYC ESE looping to the transcriptional stat site of MYC was dependent on EBNAs. Deleting MYC ESEs greatly reduced MYC expression and LCL growth. EBNA3A/3C altered CDKN2A/B spatial organization to suppress senescence. EZH2 inhibition decreased the looping at the CDKN2A/B loci and reduced LCL growth. This study provides a comprehensive view of the spatial organization of chromatin during EBV-driven cellular transformation.

爱泼斯坦-巴尔病毒（EBV）将B细胞转化为持续增殖的淋巴母细胞样细胞系（LCL），这是与EBV相关的癌症的实验模型。EBV核抗原（EBNA）和LMP1是EBV转录调节因子，对LCL的建立，增殖和存活至关重要。从LCL的3D基因组组织图开始，我们构建了一个包含1,992个病毒/细胞基因和增强子的综合EBV规则组。LCL生长必需的基因中约有30％与EBV增强子相关。删除EBNA2位点显着降低了其靶基因表达。附加的EBV超级增强器（ESE）目标包括MCL1，IRF4和EBF。MYC ESE循环到MYC的转录统计位点取决于EBNA。删除MYC ESE会大大降低MYC表达和LCL生长。EBNA3A / 3C改变了CDKN2A / B的空间组织以抑制衰老。EZH2抑制减少了CDKN2A / B基因座处的环并降低了LCL的生长。这项研究提供了在EBV驱动的细胞转化过程中染色质的空间组织的全面视图。