Formation of endonuclease III-sensitive sites as a consequence of oxygen radical attack on DNA.
Exposure of the plasmid pBR 322 to the aerobic xanthine oxidase reaction introduced single strand scissions and endonuclease III-sensitive sites. The latter may be residues of thymine glycol. Both forms of DNA damage were completely prevented by superoxide dismutase or catalase, whereas bovine serum albumin was much less effective. Mannitol and benzoate, added as scavengers of HO., and desferrioxamine or diethylene triamine pentaacetate, added to sequester Fe(III), also protected. These results indicate a metal-catalyzed interaction of O2- with H2O2, which produces HO. which, in turn, causes DNA strand scission and oxidation of thymine residues to thymine glycol. Plasmid isolated from aerobically-incubated cells contained more strand scissions and endonuclease III-sensitive sites than did plasmid from anaerobically-incubated cells, and a low molecular weight scavenger of O2- prevented the damage seen with the aerobic cells. Genetic defects in AP endonucleases rendered E. coli more susceptible to the dioxygen-dependent lethality of plumbagin, which mediates O2- production. Similarly, plasmid DNA, within the endonuclease-deficient cells, exhibited more strand scissions and endonuclease III-sensitive sites upon aerobic exposure to plumbagin than did endonuclease-sufficient cells, and a low molecular weight scavenger of O2- was protective. These results are consistent with the conclusions that strand scissions and formation of endonuclease III-sensitive sites are among the consequences of exposure of DNA to O2- plus H2O2, both in vitro and in vivo.
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Department of Biochemistry, Duke University Medical Center, Durham, NC 27710.
Fridovich
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D002374:Catalase / Q000494:pharmacology; D004249:DNA Damage; D004260:DNA Repair; D004269:DNA, Bacterial / Q000187:drug effects*; D043224:Deoxyribonuclease (Pyrimidine Dimer); D004706:Endodeoxyribonucleases / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D005609:Free Radicals; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006878:Hydroxides / Q000494:pharmacology; D017665:Hydroxyl Radical; D008345:Manganese / Q000494:pharmacology; D009285:Naphthoquinones / Q000494:pharmacology; D010100:Oxygen / Q000494:pharmacology*; D010957:Plasmids; D013482:Superoxide Dismutase / Q000494:pharmacology; D013481:Superoxides / Q000494:pharmacology
D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D005609:Free Radicals; D006878:Hydroxides; D009285:Naphthoquinones; D013481:Superoxides; D017665:Hydroxyl Radical; D008345:Manganese; D006861:Hydrogen Peroxide; D002374:Catalase; D013482:Superoxide Dismutase; D004706:Endodeoxyribonucleases; D043224:Deoxyribonuclease (Pyrimidine Dimer); C106403:NTH protein, E coli; D010100:Oxygen; C014758:plumbagin