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Accepted (AM)Open Access (License Unspecified), Open
Journal article
A versatile new tool to quantify abasic sites in DNA and inhibit base excision repair
DNA repair, v 27, pp 9-18
Mar 2015
PMCID: PMC4336807
PMID: 25616257
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
•A new alkoxyamine that reacts with abasic sites in DNA.•Bioorthogonal click chemistry.•Labeling of abasic sites with biotin or fluorophores.•Higher sensitivity and better biocompatibility than aldehyde-reactive probe.•Inhibition of AP endonuclease 1.•Better killing in combination with methyl methanesulfonate than methoxyamine.
A number of endogenous and exogenous agents, and cellular processes create abasic (AP) sites in DNA. If unrepaired, AP sites cause mutations, strand breaks and cell death. Aldehyde-reactive agent methoxyamine reacts with AP sites and blocks their repair. Another alkoxyamine, ARP, tags AP sites with a biotin and is used to quantify these sites. We have combined both these abilities into one alkoxyamine, AA3, which reacts with AP sites with a better pH profile and reactivity than ARP. Additionally, AA3 contains an alkyne functionality for bioorthogonal click chemistry that can be used to link a wide variety of biochemical tags to AP sites. We used click chemistry to tag AP sites with biotin and a fluorescent molecule without the use of proteins or enzymes. AA3 has a better reactivity profile than ARP and gives much higher product yields at physiological pH than ARP. It is simpler to use than ARP and its use results in lower background and greater sensitivity for AP site detection. We also show that AA3 inhibits the first enzyme in the repair of abasic sites, APE-1, to about the same extent as methoxyamine. Furthermore, AA3 enhances the ability of an alkylating agent, methylmethane sulfonate, to kill human cells and is more effective in such combination chemotherapy than methoxyamine.
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Details
- Title
- A versatile new tool to quantify abasic sites in DNA and inhibit base excision repair
- Creators
- Shanqiao Wei - Wayne State UniversitySophia Shalhout - Wayne State UniversityYoung-Hoon Ahn - Wayne State UniversityAshok S. Bhagwat - Wayne State University
- Publication Details
- DNA repair, v 27, pp 9-18
- Publisher
- Elsevier
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- College of Arts and Sciences; Chemistry; Drexel University
- Web of Science ID
- WOS:000350941600002
- Scopus ID
- 2-s2.0-84921513180
- Other Identifier
- 991020099677904721
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InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Genetics & Heredity
- Toxicology