Department of Medicine, Brigham and Women's Hospital, and Department of Microbiology and Immunobiology, Harvard University, Boston, MA 02115.
Proc Natl Acad Sci U S A. 2013 Nov 12;110(46):18537-42. doi: 10.1073/pnas.1317608110. Epub 2013 Oct 28.
Epstein-Barr virus (EBV) nuclear antigens EBNALP (LP) and EBNA2 (E2) are coexpressed in EBV-infected B lymphocytes and are critical for lymphoblastoid cell line outgrowth. LP removes NCOR and RBPJ repressive complexes from promoters, enhancers, and matrix-associated deacetylase bodies, whereas E2 activates transcription from distal enhancers. LP ChIP-seq analyses identified 19,224 LP sites of which ~50% were ± 2 kb of a transcriptional start site. LP sites were enriched for B-cell transcription factors (TFs), YY1, SP1, PAX5, BATF, IRF4, ETS1, RAD21, PU.1, CTCF, RBPJ, ZNF143, SMC3, NFκB, TBLR, and EBF. E2 sites were also highly enriched for LP-associated cell TFs and were more highly occupied by RBPJ and EBF. LP sites were highly marked by H3K4me3, H3K27ac, H2Az, H3K9ac, RNAPII, and P300, indicative of activated transcription. LP sites were 29% colocalized with E2 (LP/E2). LP/E2 sites were more similar to LP than to E2 sites in associated cell TFs, RNAPII, P300, and histone H3K4me3, H3K9ac, H3K27ac, and H2Az occupancy, and were more highly transcribed than LP or E2 sites. Gene affected by CTCF and LP cooccupancy were more highly expressed than genes affected by CTCF alone. LP was at myc enhancers and promoters and of MYC regulated ccnd2, 23 med complex components, and MYC regulated cell survival genes, igf2r and bcl2. These data implicate LP and associated TFs and DNA looping factors CTCF, RAD21, SMC3, and YY1/INO80 chromatin-remodeling complexes in repressor depletion and gene activation necessary for lymphoblastoid cell line growth and survival.
爱泼斯坦-巴尔病毒 (EBV) 核抗原 EBNALP (LP) 和 EBNA2 (E2) 在 EBV 感染的 B 淋巴细胞中共同表达，对于淋巴母细胞系的生长至关重要。LP 从启动子、增强子和基质相关去乙酰化酶体中去除 NCOR 和 RBPJ 抑制复合物，而 E2 则从远端增强子激活转录。LP ChIP-seq 分析鉴定了 19,224 个 LP 位点，其中约 50%位于转录起始位点的 ± 2 kb 范围内。LP 位点富含 B 细胞转录因子 (TFs)，如 YY1、SP1、PAX5、BATF、IRF4、ETS1、RAD21、PU.1、CTCF、RBPJ、ZNF143、SMC3、NFκB、TBLR 和 EBF。E2 位点也高度富集 LP 相关的细胞 TFs，并且 RBPJ 和 EBF 的占据程度更高。LP 位点高度标记有 H3K4me3、H3K27ac、H2Az、H3K9ac、RNAPII 和 P300，表明转录激活。LP 位点有 29%与 E2 (LP/E2) 共定位。LP/E2 位点与相关细胞 TFs 中的 LP 位点比 E2 位点更相似，在 RNAPII、P300 和组蛋白 H3K4me3、H3K9ac、H3K27ac 和 H2Az 占据方面也更相似，并且转录水平高于 LP 或 E2 位点。受 CTCF 和 LP 共占据影响的基因的表达水平高于仅受 CTCF 影响的基因。LP 位于 myc 增强子和启动子上，调节 MYC 调控的 ccnd2、23 med 复合物成分和 MYC 调控的细胞存活基因 igf2r 和 bcl2。这些数据表明 LP 及其相关 TF 和 DNA 环化因子 CTCF、RAD21、SMC3 和 YY1/INO80 染色质重塑复合物在淋巴母细胞系生长和存活所必需的抑制物耗竭和基因激活中起作用。
