Hypoxia impedes the formation of chromium DNA-adducts in a cell-free system.
The metabolic reduction of hexavalent chromium [Cr(VI)] in the presence of DNA generates several lesions which impede DNA replication and gene transcription. However, the relative contribution of molecular oxygen to Cr-induced genetic damage is unclear. To elucidate the role of dioxygen in Cr genotoxicity, we studied the formation of Cr-induced lesions in DNA treated with either Cr(VI) and the physiological reductant, ascorbic acid (Asc), or Cr(III), under ambient and hypoxic (<1% oxygen) conditions. We found that hypoxia did not impede the reduction of Cr(VI) by Asc throughout a 2 h treatment. In contrast, Cr-DNA binding under these conditions was reduced up to 70% by hypoxia, and a 50-90% decrease in the frequency of Cr-induced Taq polymerase-arresting DNA adducts was also observed. In the presence of Cr(VI)/Asc, formation of Cr-DNA interstrand crosslinks (ICLs) under hypoxia was 50% or less of that under ambient conditions. Kinetic studies found that hypoxia reduced the rate at which Cr interacted with DNA, but not the ultimate steady state level of Cr-DNA binding. The inhibitory effect of hypoxia on Cr(VI)/Asc genotoxicity could not be explained solely by alterations in the reactivity of intermediate Cr(V) species because Cr(III)-DNA binding and Cr(III)-induced ICL formation were also impaired by hypoxia. Moreover, Cr(V) was generated to similar levels in ambient and hypoxic reactions. Hypoxia did not affect ICL formation by the inorganic chemotherapeutic agent cisplatin, suggesting that these effects were specific for Cr(III). Taken together, these results support a role for dioxygen in facilitating the formation of Cr-DNA coordination complexes.
O'Brien
Travis J
TJ
Department of Pharmacology and Physiology, The George Washington University Medical Center, 2300 Eye Street NW, Washington, DC 20037, USA.
Mandel
H George
HG
Sugden
Kent D
KD
Komarov
Andrei M
AM
Patierno
Steven R
SR
D002474:Cell-Free System; D002478:Cells, Cultured; D002857:Chromium / Q000378:metabolism* / Q000633:toxicity; D002945:Cisplatin / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D018736:DNA Adducts / Q000378:metabolism*; D006801:Humans; D010100:Oxygen / Q000502:physiology*; D010957:Plasmids
D018736:DNA Adducts; D002857:Chromium; D004247:DNA; D002945:Cisplatin; D010100:Oxygen