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Selective Chemical Labeling of Natural T Modifications in DNA.


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Authors

Hardisty, Robyn E 
Kawasaki, Fumiko 
Sahakyan, Aleksandr B 
Balasubramanian, Shankar  ORCID logo  https://orcid.org/0000-0002-0281-5815

Abstract

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.

Description

Keywords

Base Sequence, DNA, Models, Molecular, Nucleic Acid Conformation, Oligodeoxyribonucleotides, Pentoxyl, Thymine, Uracil

Journal Title

J Am Chem Soc

Conference Name

Journal ISSN

0002-7863
1520-5126

Volume Title

137

Publisher

American Chemical Society (ACS)
Sponsorship
Engineering and Physical Sciences Research Council (EP/K039520/1)
Wellcome Trust (099232/Z/12/Z)
R.E.H. is supported by The University of Cambridge, F.K. is supported by the Wellcome Trust, and A.B.S. is supported by the Herchel Smith Fund. The Balasubramanian group is core- funded by a Wellcome Trust Senior Investigator Award and by Cancer Research UK. Departmental NMR facilities are supported by EPSRC grant EP/K039520/1.