The mechanism of the nitric oxide-mediated enhancement of tert-butylhydroperoxide-induced DNA single strand breakage.
1. Caffeine (Cf) enhances the DNA cleavage induced by tert-butylhydroperoxide (tB-OOH) in U937 cells via a mechanism involving Ca2+-dependent mitochondrial formation of DNA-damaging species (Guidarelli et al., 1997b). Nitric oxide (NO) is not involved in this process since U937 cells do not express the constitutive nitric oxide synthase (cNOS). 2. Treatment with the NO donors S-nitroso-N-acetyl-penicillamine (SNAP, 10 microM), or S-nitrosoglutathione (GSNO, 300 microM), however, potentiated the DNA strand scission induced by 200 microM tB-OOH. The DNA lesions generated by tB-OOH alone, or combined with SNAP, were repaired with superimposable kinetics and were insensitive to anti-oxidants and peroxynitrite scavengers but suppressed by iron chelators. 3. SNAP or GSNO did not cause mitochondrial Ca2+ accumulation but their enhancing effects on the tB-OOH-induced DNA strand scission were prevented by ruthenium red, an inhibitor of the calcium uniporter of mitochondria. Furthermore, the enhancing effects of both SNAP and GSNO were identical to and not additive with those promoted by the Ca2+-mobilizing agents Cf or ATP. 4. The SNAP- or GSNO-mediated enhancement of the tB-OOH-induced DNA cleavage was abolished by the respiratory chain inhibitors rotenone and myxothiazol and was not apparent in respiration-deficient cells. 5. It is concluded that, in cells which do not express the enzyme cNOS, exogenous NO enhances the accumulation of DNA single strand breaks induced by tB-OOH via a mechanism involving inhibition of complex III.
Guidarelli
A
A
Istituto di Farmacologia e Farmacognosia and Centro di Farmacologia Oncologica Sperimentale, Universitá di Urbino, Italy.
Clementi
E
E
Sciorati
C
C
Cantoni
O
O
D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D003575:Cytochromes c1 / Q000378:metabolism; D004249:DNA Damage; D004277:DNA, Single-Stranded / Q000187:drug effects* / Q000378:metabolism; D004579:Electron Transport; D006801:Humans; D008928:Mitochondria / Q000378:metabolism; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000378:metabolism; D009566:Nitrates / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism* / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000096:biosynthesis; D052248:Nitric Oxide Synthase Type I; D010396:Penicillamine / Q000031:analogs & derivatives / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D020298:U937 Cells; D014451:Ubiquinone / Q000378:metabolism; D020122:tert-Butylhydroperoxide / Q000494:pharmacology*
D004277:DNA, Single-Stranded; D009566:Nitrates; C110881:S-nitro-N-acetylpenicillamine; D014451:Ubiquinone; C039642:peroxynitric acid; D009569:Nitric Oxide; D002110:Caffeine; D003575:Cytochromes c1; D020122:tert-Butylhydroperoxide; C496320:NOS1 protein, human; D019001:Nitric Oxide Synthase; D052248:Nitric Oxide Synthase Type I; D016660:NAD(P)H Dehydrogenase (Quinone); D010396:Penicillamine; D002118:Calcium