Abstract
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), Mn(II) and Ca(II)]. Structurally related flavonoids, rutin, galangin, apigenin and fisetin were effective or less effecive than quercetin in causing DNA breakage. In the case of the quercetin-Cu(II) reaction, Cu(I) was shown to be essential intermediate by using the Cu(1)-sequestering reagent, bathocuproine. By using Job plots we established that, in the absence of DNA, five Cu(II) ions were 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). The strand scission reaction was shown to account for the biological activity of quercetin as assayed by bacteriophage inactivation. From these data we propose a mechanism for the DNA strand scission reaction of quercetin and related flavonoids.
Similar content being viewed by others
References
Ames BN: Dietary carcinogens and anticarcinogens. Science 221: 1256–1264, 1983
Herman K: Flavonoids and flavones in food plants, a review. J Food Sci Technol 11: 433–448, 1976
Nagao M, Sugimura T, Matsushima T: Environmental mutagens and carcinogens. Ann Rev Genet 12: 117–159, 1978
Brown JP: A review of genetic effects of naturally occurring flavonoids, anthraquinones and related compounds. Mutat Res 75: 243–277, 1980
Hirono I, Ueno I, Hosaka S, Takanashi H, Matsushima T, Sugimura T, Natori S: Carcinogenicity examination of quercetin and rutin in ACI rats. Cancer Lett 13: 15–21, 1981
Pamukcu AM, Yalciner S, Hatcher JF, Bryan JT: Quercetin a rat intestinal and bladder carcinogen present in bracken fern (pteridium aquilinum). Cancer Res 40: 3468–3472, 1980
Watson WAF: The mutagenic activity of quercetin and kaempferol in Drosophila melanogaster. Mutat Res 103: 145–147, 1982
Hatcher JF, Bryan JT: Factors affecting the mutagenic activity of quercetin in Salmonella typhimurium TA 98: metal ions, antioxidants and pH. Mutat Res 149: 13–23, 1986
Vlietinck AJ, Vanden Berghe DA, Haemers A: Present status and prospects of flavonoids as antiviral agents. In: V Cody, E Middleton, JB Harborne (eds.) Plant Flavonoids in Biology and Medicine II: biochemical, cellular and medicinal properties, Alan R Liss, New York, 1987, pp 283–299
Bracke ME, Pestel GD, Castronova V, Vyncke B, Foidart JM, Vakaet LCA, Mareel MM: Flavonoids inhibit malignant tumor invasion in vitro. In: V Cody, E Middleton, JB Harborne (eds.) Plant Flavonoids in Biology and Medicine II: biochemical, cellular and medicinal properties, Alan R Liss, New York, 1987, pp 219–223
Takahama U: O2 −-dependent and independent photo-oxidation of quercetin in the presence and absence of riboflavin and effects of ascorbate on the photo-oxidation. Photochem Photobiol 42: 89–91, 1985
Hadi SM, Shahabuddin, Rahman A: Specificity of interaction of furfural with DNA. Mutat Res 225: 102–106, 1989
Wong A, Haung CH, Crooke ST: Deoxyribonucleic acid breakage produced by 4′(9-acridinylamino)methanesulfon-m-anisidide and copper: role of cuprous ion and oxygen free radicals. Biochemistry 23: 2939–2945, 1984
Maniatis T, Fritsch E, Sambrook J: Molecular Cloning: a laboratory manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1982, pp 80–85
Fazal F, Rahman A, Greensill J, Ainley K, Hadi SM, Parish JH: Strand scission in DNA by quercetin and Cu(II): identification of free radical intermediate and biological consequences of scission. Carcinogenesis 11: 2005–2008, 1990
Kohen R, Syzf M, Chevion M: Quantitation of single and double strand breaks in vitro and in vivo. Anal Biochem 154: 485–491, 1986
Wong A, Haung CH, Crooke ST: Mechanism of deoxyribonucleic acid breakage induced by 4′(9-acridinylamino)methanesul-fon-m-anisidide and copper: role of cuprous ion and oxygen free radicals, Biochemistry 23: 2946–2952, 1984
Rahman A, Shahabuddin, Hadi SM, Parish JH, Ainley K: Strand scission in DNA induced by quercetin and Cu(II): role of Cu(I) and oxygen free radicals. Carcinogenesis 10: 1833–1839, 1989
Macgregor TT, Jurd L: Mutagenicity of flavonoids: structural requirements for mutagenic activity in Salmonella typhimurium. Mutat Res 54: 297–309, 1978
Graham DR, Marshall LE, Reich KE, Sigman DS: Cleavage of DNA by coordination complexes. Superoxide formation in the oxidation of 1,10-phenanthroline cuprous complexes by oxygen: relevance to DNA cleavage reaction. J Am Chem Soc 102: 5419–5412, 1981
Dreyer GB, Dervan PB: Sequence-specific cleavage of single stranded DNA. Proc Natl Acad Soc USA 82: 968–972, 1985
Gutteridge JMC, Halliwell B: The role of superoxide and hydroxyl radicals in the degradation of DNA and deoxyribose by copper phenanthroline complexes. Biochem Pharmacol 31: 2801–2805, 1982
Ehrenfeld GM, Shipley JB, Heimbrook DC, Sugiyama H, Long EC, Van Boom JH, Vander Marel GA, Oppenheimer NJ, Hecht SM: Copper dependent cleavage of DNA by bleomycin. Biochemistry 26: 931–942, 1987
Eliot H, Gianni L, Myers C: Oxidative destruction of DNA by adriamycin-iron complexes. Biochemistry 23: 928–936, 1984
Quinlan GJ, Gutteridge JMC: Oxygen radical damage to DNA by rifamycin and copper ions. Biochem Pharmacol 36: 3629–3633, 1987
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Rahman, A., Fazal, F., Greensill, J. et al. Strand scission in DNA induced by dietary flavonoids: role of Cu(I) and oxygen free radicals and biological consequences of scission. Mol Cell Biochem 111, 3–9 (1992). https://doi.org/10.1007/BF00229567
Issue Date:
DOI: https://doi.org/10.1007/BF00229567