Selective Chemical Labeling of Natural T Modifications in DNA.
Selective Chemical Labeling of Natural T Modifications in DNA.
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DOI：
10.1021/jacs.5b03730
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发表时间：
2015-07-29
影响因子：
15
通讯作者：
Balasubramanian S
中科院分区：
文献类型：
--
作者：
Hardisty RE;Kawasaki F;Sahakyan AB;Balasubramanian S
我们提出了一种化学方法来选择性地标记和富集胸腺嘧啶修饰，5-甲酰尿嘧啶（5-FU）和5-羟甲基尿嘧啶（5-hmU），天然存在于DNA中。固有的反应性差异，使我们能够标记5-FU化学选择性超过其C修饰对应物，5-甲酰胞嘧啶（5-fC）。我们通过从头算量子力学计算合理化了5-fU相比5-fC的增强反应性。我们利用这种化学标记反应来提供从含有5-fC或无修饰的池中富集含有5-fU的DNA的概念证明。我们进一步证明，5-hmU可以被化学氧化为5-FU，提供了一种富集5-hmU的策略。这些方法将使5-fU和5-hmU在基因组DNA中的定位成为可能，以深入了解它们在生物学中的功能作用和动态。
We present a chemical method to selectively tag and enrich thymine modifications, 5-formyluracil (5-fU) and 5-hydroxymethyluracil (5-hmU), found naturally in DNA. Inherent reactivity differences have enabled us to tag 5-fU chemoselectively over its C modification counterpart, 5-formylcytosine (5-fC). We rationalized the enhanced reactivity of 5-fU compared to 5-fC via ab initio quantum mechanical calculations. We exploited this chemical tagging reaction to provide proof of concept for the enrichment of 5-fU containing DNA from a pool that contains 5-fC or no modification. We further demonstrate that 5-hmU can be chemically oxidized to 5-fU, providing a strategy for the enrichment of 5-hmU. These methods will enable the mapping of 5-fU and 5-hmU in genomic DNA, to provide insights into their functional role and dynamics in biology.