A reliable assessment of 8-oxo-2-deoxyguanosine levels in nuclear and mitochondrial DNA using the sodium iodide method to isolate DNA

Michelle L. Hamilton, Zhong Mao Guo, Clinton D. Fuller, Holly Van Remmen, Walter F. Ward, Steven N. Austad, Dean A. Troyer, Ian Thompson, Arlan Richardson

Research output: Contribution to journalArticlepeer-review

160 Scopus citations

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-deoxyguanosine (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/105 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 γ-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 γ-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.

Original languageEnglish (US)
Pages (from-to)2117-2126
Number of pages10
JournalNucleic acids research
Volume29
Issue number10
DOIs
StatePublished - May 15 2001

ASJC Scopus subject areas

  • Genetics

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