Document Type
Article
Publication Date
2012
Publication Title
Nucleic Acids Research
Abstract
The misreplication of damaged DNA is an important biological process that produces numerous adverse effects on human health. This report describes the synthesis and characterization of a non-natural nucleotide, designated 3-ethynyl-5-nitroindolyl-2′-deoxyriboside triphosphate (3-Eth-5-NITP), as a novel chemical reagent that can probe and quantify the misreplication of damaged DNA. We demonstrate that this non-natural nucleotide is efficiently inserted opposite an abasic site, a commonly formed and potentially mutagenic non-instructional DNA lesion. The strategic placement of the ethynyl moiety allows the incorporated nucleoside triphosphate to be selectively tagged with an azide-containing fluorophore using ‘click’ chemistry. This reaction provides a facile way to quantify the extent of nucleotide incorporation opposite non-instructional DNA lesions. In addition, the incorporation of 3-Eth-5-NITP is highly selective for an abasic site, and occurs even in the presence of a 50-fold molar excess of natural nucleotides. The biological applications of using 3-Eth-5-NITP as a chemical probe to monitor and quantify the misreplication of non-instructional DNA lesions are discussed.
Recommended Citation
Motea, Edward A.; Lee, Irene; and Berdis, Anthony J., "Development of A ‘Clickable’ Non-Natural Nucleotide to Visualize The Replication of Non-Instructional DNA Lesions" (2012). Chemistry Faculty Publications. 183.
https://engagedscholarship.csuohio.edu/scichem_facpub/183
DOI
10.1093/nar/gkr980
Version
Publisher's PDF
Volume
40
Issue
5
