Chromatin features are widely used for genome-scale mapping of enhancers. However, discriminating active enhancers from other cis-regulatory elements, predicting enhancer strength and identifying their target genes is challenging. Here we establish histone H2B N-terminus multisite lysine acetylation (H2BNTac) as a signature of active enhancers. H2BNTac prominently marks candidate active enhancers and a subset of promoters and discriminates them from ubiquitously active promoters. Two mechanisms underlie the distinct H2BNTac specificity: (1) unlike H3K27ac, H2BNTac is specifically catalyzed by CBP/p300; (2) H2A–H2B, but not H3–H4, are rapidly exchanged through transcription-induced nucleosome remodeling. H2BNTac-positive candidate enhancers show a high validation rate in orthogonal enhancer activity assays and a vast majority of endogenously active enhancers are marked by H2BNTac and H3K27ac. Notably, H2BNTac intensity predicts enhancer strength and outperforms current state-of-the-art models in predicting CBP/p300 target genes. These findings have broad implications for generating fine-grained enhancer maps and modeling CBP/p300-dependent gene regulation.

染色质特征广泛用于增强子的基因组规模作图。然而，将活性增强剂与其他顺式区分开来-调节元件，预测增强子强度和识别它们的目标基因是具有挑战性的。在这里，我们建立组蛋白 H2B N 末端多位点赖氨酸乙酰化 (H2BNTac) 作为活性增强子的标志。H2BNTac 显着标记候选活性增强子和一部分启动子，并将它们与无处不在的活性启动子区分开来。两种机制构成了独特的 H2BNTac 特异性：(1) 与 H3K27ac 不同，H2BNTac 由 CBP/p300 特异性催化；(2) H2A–H2B，而非 H3–H4，通过转录诱导的核小体重塑快速交换。H2BNTac 阳性候选增强子在正交增强子活性测定中显示出高验证率，并且绝大多数内源活性增强子由 H2BNTac 和 H3K27ac 标记。尤其，H2BNTac 强度预测增强子强度，并且在预测 CBP/p300 靶基因方面优于当前最先进的模型。这些发现对于生成细粒度的增强子图谱和模拟 CBP/p300 依赖性基因调控具有广泛的意义。