Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, 5-1-5 Kashiwanoha, Kashiwashi, Chiba 277-8562, Japan.
DNA Res. 2011 Oct;18(5):379-92. doi: 10.1093/dnares/dsr025. Epub 2011 Aug 9.
Using ChIP Seq, we identified 556 and 467 putative STAT6 target sites in the Burkitt's lymphoma cell line Ramos and in the normal lung epithelial cell line BEAS2B, respectively. We also examined the positions and expression of transcriptional start sites (TSSs) in these cells using our TSS Seq method. We observed that 44 and 132 genes in Ramos and BEAS2B, respectively, had STAT6 binding sites in proximal regions of their previously reported TSSs that were up-regulated at the transcriptional level. In addition, 406 and 109 of the STAT6 target sites in Ramos and BEAS2B, respectively, were located in proximal regions of previously uncharacterized TSSs. The target genes identified in Ramos and BEAS2B cells in this study and in Th2 cells in previous studies rarely overlapped and differed in their identity. Interestingly, ChIP Seq analyses of histone modifications and RNA polymerase II revealed that chromatin formed an active structure in regions surrounding the STAT6 binding sites; this event also frequently occurred in different cell types, although neither STAT6 binding nor TSS induction was observed. The rough landscape of STAT6-responsive sites was found to be shaped by chromatin structure, but distinct cellular responses were mainly mediated by distinct sets of transcription factors.
使用 ChIP Seq,我们分别在 Burkitt 淋巴瘤细胞系 Ramos 和正常肺上皮细胞系 BEAS2B 中鉴定了 556 个和 467 个假定的 STAT6 靶位点。我们还使用我们的 TSS Seq 方法检查了这些细胞中转录起始位点 (TSS) 的位置和表达。我们观察到,在 Ramos 和 BEAS2B 中,分别有 44 个和 132 个基因在其先前报道的 TSS 的近端区域具有 STAT6 结合位点,这些基因在转录水平上上调。此外,在 Ramos 和 BEAS2B 中,分别有 406 个和 109 个 STAT6 靶位点位于先前未表征的 TSS 的近端区域。在这项研究的 Ramos 和 BEAS2B 细胞以及先前研究中的 Th2 细胞中鉴定的靶基因很少重叠,其身份也不同。有趣的是,组蛋白修饰和 RNA 聚合酶 II 的 ChIP Seq 分析表明,染色质在 STAT6 结合位点周围的区域形成了一个活跃的结构;这种事件也经常发生在不同的细胞类型中,尽管既没有观察到 STAT6 结合也没有观察到 TSS 诱导。STAT6 反应性位点的大致景观被发现是由染色质结构塑造的,但不同的细胞反应主要是由不同的转录因子集介导的。