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- Title
A reliable assessment of 8-oxo-2-deoxyguanosine levels in nuclear and mitochondrial DNA using the sodium iodide method to isolate DNA.
- Authors
Hamilton, Michelle L.; Guo, ZhongMao; Fuller, Clinton D.; Van Remmen, Holly; Ward, Walter F.; Austad, Steven N.; Troyer, Dean A.; Thompson, Ian; Richardson, Arlan
- Abstract
Abstract A major controversy in the area of DNA biochemistry concerns the actual in vivo levels of oxidative damage in DNA. We show here that 8-oxo-2-deoxy-guanosine (oxo8dG) generation during DNA isolation is eliminated using the sodium iodide (Nal) isolation method and that the level of oxo8dG in nuclear DNA (nDNA) is almost one-hundredth of the level obtained using the classical phenol method. We found using Nal that the ratio of oxo8dG/10[sup 5] deoxyguanosine (dG) in nDNA isolated from mouse tissues ranged from 0.032 +/- 0.002 for liver to 0.015 +/- 0.003 for brain. We observed a significant increase (10-fold) in oxo8dG in nDNA isolated from liver tissue after 2 Gy of gamma-irradiation when Nal was used to isolate DNA. The turnover of oxo8dG in nDNA was rapid, e.g. disappearance of oxo8dG in the mouse liver in vivo after gamma-irradiation had a half-life of 11 min. The levels of oxo8dG in mitochondrial DNA isolated from liver, heart and brain were 6-, 16- and 23-fold higher than nDNA from these tissues. Thus, our results showed that the steady-state levels of oxo8dG in mouse tissues range from 180 to 360 lesions in the nuclear genome and from one to two lesions in 100 mitochondrial genomes.
- Subjects
DNA damage; BIOCHEMISTRY; MITOCHONDRIAL DNA
- Publication
Nucleic Acids Research, 2001, Vol 29, Issue 10, p2117
- ISSN
0305-1048
- Publication type
Article
- DOI
10.1093/nar/29.10.2117