Interaction of antimutagenic 1,4-dihydropyridine AV-153-Na with DNA and DNA-damaging molecules and its impact on DNA repair activity
- Published
- Accepted
- Subject Areas
- Biophysics, Drugs and Devices
- Keywords
- 4-dihydropyridines, AV-153-Na, DNA repair, DNA binding
- Copyright
- © 2017 Leonova et al.
- Licence
- This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ Preprints) and either DOI or URL of the article must be cited.
- Cite this article
- 2017. Interaction of antimutagenic 1,4-dihydropyridine AV-153-Na with DNA and DNA-damaging molecules and its impact on DNA repair activity. PeerJ Preprints 5:e3317v1 https://doi.org/10.7287/peerj.preprints.3317v1
Abstract
1,4-dihydropyridines (1,4-DHP) possess important biochemical and pharmacological properties, including antioxidant and antimutagenic activities. Interaction of some 1,4-DHP with DNA was recently reported. AV-153-Na, an antimutagenic and DNA-repair-enhancing compound appeared to be able to interact with DNA by intercalation. The aim of the current study was to characterize DNA’s capacity for the binding of AV-153-Na, and using different approaches, to test intracellular distribution of the compound, to test the ability of the compound to scavenge peroxynitrite and hydroxyl radical and to assess the ability of the compound to modify the activity of DNA repair enzymes. The DNA binding activity of AV-153-Na was determined by means of fluorescence assay. Titration of the AV-153-Na solutions with DNA gradually increased fluorescence of the solution, indicating direct interactions of the molecule with DNA. AV-153-Na quenched the fluorescence of ethidium bromide and DNA complex, which points to intercalation binding mode. Binding via intercalation was confirmed by means of cyclic voltammetry and circular dichroism spectroscopy. The compound could interact with the four DNA bases in vitro, manifesting a higher affinity to guanine. Some ability to scavenge hydroxyl radical by AV-153-Na was detected by the EPR method. AV-153-Na turned out to be incapable of reacting chemically with peroxynitrite. However, AV-153-Na effectively decreased DNA damage produced by peroxynitrite in cultured HeLa cells. The effects of AV-153-Na on the activity of DNA repair enzymes were tested using Glyco-SPOT and ExSy-SPOT assays. The Glyco-SPOT test essentially revealed an inhibition by AV-153-Na of the enzymes involved thymine glycol repair. Results with ExSy-SPOT chip indicate that AV-153-Na significantly stimulates excision/synthesis repair of 8-oxoguanine (8-oxoG), abasic sites (AP sites) and alkylated bases. Laser confocal scanning fluorescence microscopy demonstrated that within the cells AV-153-Na was found mostly in the cytoplasm; however, a stain in nucleolus was also detected. Binding to cytoplasmic structures might occur due to high affinity of the compound to protein, revealed by fluorescence spectroscopy titration and circular dichroism. Activation of DNA repair enzymes after binding to DNA appears to be the basis for the antimutagenic effects of AV-153-Na.
Author Comment
This is preprint submission to PeerJ Preprints
Supplemental Information
Effects of AV-153-Na (3h incubation) against peroxinitrite
Figure 9 (B)
Effects of AV-153-Na on the activity of DNA repair enzymes
Figures 10 and 11
Effect of AV-153-Na (45 min) against peroxynitrite
Figure 9 (A)
Effect of AV-154-Na (45 min) agaist peroxynitrite
Figure 9 (C)