Selective Chemical Labeling of Natural T Modifications in DNA

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• 作者：Robyn E. Hardisty ; Fumiko Kawasaki ; Aleksandr B. Sahakyan ; Shankar Balasubramanian
• 刊名：Journal of the American Chemical Society
• 出版年：2015
• 出版时间：July 29, 2015
• 年：2015
• 卷：137
• 期：29
• 页码：9270-9272
• 全文大小：239K
• ISSN：1520-5126

文摘

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.