Uracil in DNA can be generated by cytosine deamination or dUMP misincorporation; however, its distribution in the human genome is poorly understood. Here we present a selective labeling and pull-down technology for genome-wide uracil profiling and identify thousands of uracil peaks in three different human cell lines. Surprisingly, uracil is highly enriched at the centromere of the human genome. Using mass spectrometry, we demonstrate that human centromeric DNA contains a higher level of uracil. We also directly verify the presence of uracil within two centromeric uracil peaks on chromosomes 6 and 11. Moreover, centromeric uracil is preferentially localized within the binding regions of the centromere-specific histone CENP-A and can be excised by human uracil-DNA glycosylase UNG. Collectively, our approaches allow comprehensive analysis of uracil in the human genome and provide robust tools for mapping and future functional studies of uracil in DNA.

DNA中的尿嘧啶可通过胞嘧啶脱氨或dUMP错掺而产生；然而，人们对其在人类基因组中的分布了解甚少。在这里，我们介绍了用于全基因组尿嘧啶分析的选择性标记和下拉技术，并在三种不同的人类细胞系中鉴定了数千个尿嘧啶峰。令人惊讶地，尿嘧啶在人类基因组的着丝粒高度富集。使用质谱法，我们证明了人类着丝粒DNA含有更高水平的尿嘧啶。我们还直接验证了染色体6和11上两个着丝粒尿嘧啶峰内尿嘧啶的存在。此外，着丝粒尿嘧啶优先位于着丝粒特异性组蛋白CENP-A的结合区内，并且可以被人尿嘧啶DNA糖基化酶UNG切除。 。总的来说，