Cobalt(II) inhibits the incision and the polymerization step of nucleotide excision repair in human fibroblasts.
Compounds of cobalt are carcinogenic to experimental animals, but the mutagenicity in mammalian cells in culture is rather weak. In contrast, cobalt(II) has been shown to inhibit the removal of DNA damage induced by UVC light, indicating an interference with cellular DNA repair processes. In the present study it was investigated which step of the nucleotide excision repair is affected by cobalt(II) and which mechanisms are involved. In this context, the effect of non-cytotoxic cobalt(II) concentrations on the induction as well as on the repair of UVC-induced DNA lesions has been examined in human fibroblasts by using the alkaline unwinding technique under various conditions. Cobalt(II) concentrations as low as 50 microM inhibit the incision as well as the polymerization step. In contrast, the ligation of repair patches is not disturbed by this metal. By combining the alkaline unwinding technique with the repair enzyme T4 endonuclease V, it is demonstrated that the incision at the site of cyclobutane pyrimidine dimers is affected at concentrations of 150 microM and higher. As one mode of action, the competition with essential magnesium(II) ions by cobalt(II) ions could be identified.
Kasten
U
U
University of Bremen, Department of Biology and Chemistry, Germany.
Mullenders
L H
LH
Hartwig
A
A
D003035:Cobalt / Q000494:pharmacology*; D004260:DNA Repair / Q000187:drug effects*; D005347:Fibroblasts / Q000528:radiation effects; D006801:Humans; D011108:Polymers / Q000378:metabolism*; D011740:Pyrimidine Dimers
D011108:Polymers; D011740:Pyrimidine Dimers; D003035:Cobalt