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

Academic Article

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.
  • Authors

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    Digital Object Identifier (doi)

    Author List

  • Hamilton ML; Guo ZM; Fuller CD; Van Remmen H; Ward WF; Austad SN; Troyer DA; Thompson I; Richardson A
  • Start Page

  • 2117
  • End Page

  • 2126
  • Volume

  • 29
  • Issue

  • 10