Advertisement

Hydroxyl Free Radical Mediated Strand Breaks in PBR322 Plasmid DNA

  • E. Schneider
  • J. J. Watson
  • R. A. Floyd

Abstract

Oxygen free radicals react with and cause damage to DNA. Hydroxyl free radicals (·OH) add to DNA to yield several products, one of which is 8-hydroxydeoxyguanine (1). Digestion of DNA to the nucleoside level yields 8-hydroxydeoxyguanosine (8-OHdG) which can be very sensitively quantitated (to 20 femtomoles) by HPLC-electrochemical detection (2,3). The potential significance of the 8-OHdG adduct in living systems has been demonstrated by its presence in the DNA of human granulocytes following TPA-stimulated oxidative damage (3). The amount of 8-OHdG present in the granulocyte DNA was decreased by the addition of superoxide dismutase and/or catalase, thus emphasizing that oxygen free radical intermediates are the responsible agents (3).

Keywords

Double Strand Break Hydroxyl Free Radical Supercoiled Plasmid Strand Scission Cetylpyridinium Bromide 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    H. Kasai and S. Nishimura, Hydroxylation of deoxyguanosine at the C-8 position by ascorbic acid and other reducing agents. Nucleic Acids Res. 12, 2137–2145 (1984).PubMedCrossRefGoogle Scholar
  2. 2.
    R.A. Floyd, J.J. Vatson, P.K. Wong, D.H. Altmiller, R.C. Rickard, Hydroxyl free radical adduct of deoxyguanosine: sensitive detection and mechanisms of formation. Free Radical Res. Commun. 1, 163–172 (1986).Google Scholar
  3. 3.
    R.A. Floyd, J.J. Watson, J. Harris, M. West, and P.K. Wong, Formation of 8-hydroxyguanosine, hydroxyl free radical adduct of DNA in granulocytes exposed to the tumor promoter, tetradeconylphoboloutate. Biochem. Biophys. Res. Commun. 137, 841–846 (1986).PubMedCrossRefGoogle Scholar
  4. 4.
    K. Brawn and I. Fridovich, DNA strand scission by enzymatically generated oxygen radicals. Arch. Biochem. Biophys. 206, 414–419 (1981).PubMedCrossRefGoogle Scholar
  5. 5.
    P. Geek and I. Nasz, Concentrated, digestible DNA after hydroxylapetite chromatography with cetylpyridinium bromide precipitation. Anal. Biochem. 135, 264–268 (1983).CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1987

Authors and Affiliations

  • E. Schneider
    • 1
  • J. J. Watson
    • 1
  • R. A. Floyd
    • 1
  1. 1.Molecular Toxicology Research GroupOklahoma Medical Research FoundationOklahoma CityUSA

Personalised recommendations