Differential effects of toxic metal compounds on the activities of Fpg and XPA, two zinc finger proteins involved in DNA repair

Abstract

Even though not mutagenic, compounds of the carcinogenic metals nickel, cadmium, cobalt and arsenic have been shown previously to inhibit nucleotide excision repair and base excision repair at low, non-cytotoxic concentrations. Since some toxic metals have high affinities for -SH groups, we used the bacterial formamidopyrimidine-DNA glycosylase (Fpg protein) and the mammalian XPA protein as models to investigate whether zinc finger structures in DNA repair enzymes are particularly sensitive to carcinogenic and/or toxic metal compounds. Concentrations of </=1 mM Ni(II), Pb(II), As(III) or Co(II) did not affect the activity of the Fpg protein significantly. In contrast, the enzyme was inhibited in a dose-dependent manner by Cd(II), Cu(II) or Hg(II), starting at concentrations of 50 microM, 5 microM and 50 nM, respectively. Simultaneous treatment with Cd(II) or Cu(II) and Zn(II) partly prevented the inhibitions, while no reversal of inhibition was observed when Zn(II) was added after Cd(II) or Cu(II). In the case of Hg(II), Zn(II) had no protective effect independent of the time of its addition; however, the enzyme activity was completely restored by glutathione. Regarding XPA, Hg(II), Pb(II) or As(III) did not diminish its binding to an UV-irradiated oligonucleotide, while Cd(II), Co(II), Cu(II) and Ni(II) reduced its DNA-binding ability. Simultaneous treatment with Zn(II) prevented largely the inhibition induced by Cd(II), Co(II), and Ni(II), but only slightly in the case of Cu(II). Collectively, the results indicate that both proteins were inhibited by Cd(II) and Cu(II), XPA was additionally inactivated by Ni(II) and Co(II), and Fpg but not XPA was strongly affected by Hg(II). Even though other mechanisms of protein inactivation cannot be completely excluded, zinc finger structures may be sensitive targets for toxic metal compounds, but each zinc finger protein has unique sensitivities.