The naturally occurring flavonoid, quercetin, in the presence of Cu(II) and molecular oxygen caused breakage of calf thymus DNA, supercoiled pBR322 plasmid DNA and single-stranded M13 phage DNA. In the case of the plasmid, the product(s) were relaxed circles or a mixture of these and linear molecules depending upon the conditions. For the breakage reaction, Cu(II) could be replaced by Fe(III) but not by other ions tested [Fe(II), Co(II), Ni(II) and Ca(II)]. Structurally related flavonoids, rutin, galangin, apigenin and fisetin, were ineffective or less effective than quercetin in causing DNA breakage. In the case of the quercetin--Cu(II) reaction, Cu(I) was shown to be an essential intermediate by using the Cu(I)-sequestering reagents, neocuproine and bathocuproine. By using Job plots we established that, in the absence of DNA, five Cu(II) ions can be reduced by one quercetin molecule; in contrast, two ions were reduced per quercetin molecule in the DNA breakage reaction. Equally neocuproine inhibited the DNA breakage reaction. The involvement of active oxygen in the reaction was established by the inhibition of DNA breakage by superoxide dismutase, iodide, mannitol, formate and catalase (the inhibition was complete in the last case). From these data we propose a mechanism for the DNA strand scission reaction of quercetin and related flavonoids.