Abstract
Pathological conditions that cause oxidative stress can affect DNA integrity. The aim of this research was to study the protective effect of spermine against DNA damage induced by an oxygen-radical generating system. Deoxyguanosine and DNA were separately dissolved in phosphate buffer and incubated for 1 h at 40°C in the presence of 50 mMH2O2/10 mM ascorbic acid. Single nucleosides and their products of oxidation were then obtained by enzymatic digestion of DNA. The compounds were separated by micellar electrokinetic capillary chromatography (MECC) with SDS-modified mobile phase and detected at 254 nm. Two major products of DNA oxidation have been identified as derivatives of deoxyguanosine with electrophoretic properties different from 8-hydroxy-2′-deoxyguanosine. When the oxidation of DNA was carried out in the presence of 0.1 mM spermine, the formation of the two by-products of deoxyguanosine was markedly reduced. On the contrary, spermine did not prevent the oxidation of deoxyguanosine alone, suggesting that the polyamine should be bound to the DNA strands to exert its antioxidative effect.
Similar content being viewed by others
References
halliwell B, Aruoma OI: DNA damage by oxygen-derived species. Its mechanism and measurement in mammalian systems. FEBS 281: 9–19, 1991
Mullaart E, Lohman PHM, Berends F, Vijg J: DNA damage metabolism and ageing. Mutat Res 237: 189–210, 1990
Takeda N, Tanamura A, Iwai T, Nakamura I, Kato M, Ohhkubo T, Noma K: Mitochondrial DNA deletion in human myocardium. Mol Cell Biochem 119: 105–108, 1993
Corral-Debrinski M, Stepien G, Shoffner JM, Lott MT, Kanter K, Wallace DC: Hypoxiemia is associated with mitochondrial DNA damage and gene induction. JAMA 266: 1812–1816, 1991
Miquel J: An integrated theory of ageing as result of mitochondrial-DNA mutation in differentiated cells. Arch Gerontol Geriatr 12: 99–117, 1991
Halliwell B, Dizdaroglu M: The measurement of oxidative damage to DNA by HPLC and GC/MS techniques. Free Rad Res Comms 16: 75–87, 1992
Richter C, Park JW, Ames BN: Normal oxidative damage to mitochondrial and nuclear DNA is extensive. Proc Natl Acad Sci USA 85: 6465–6467, 1988
Barciszewski J, Rattan SIS, Siboska GE, Otzen DE, Clark BFC: Reduction in the amount of 8-hydroxy-2′-deoxyguanosine in the DNA of SV40-transformed human fibroblasts as compared with normal cells in culture. FEBS 318: 186–188, 1993
Hayakawa M, Hattori K, Sugiyama S, Ozawa T: Age associated oxygen damage and mutations in mitochondrial DNA in human hearts. Biochem Biophys Res Communs 189: 979–985, 1992
Tabor CW, Tabor H: Polyamines. Ann Rev Biochem 53: 749–790, 1984
Igarashi K, Sakamoto I, Goto N, Kashiwagi K, Honma R, Hirose S: Interaction between polyamines and nucleic acids or phospholipids. Arch Biochem Biophys 219: 438–443, 1982
Lovaas E, Carlin G: Spermine: an anti-oxidant and anti-inflammatory agent. Free Radic Biol Med 11: 455–461, 1991
Matkovics B, Kecskemeti V, Varge SI, Novak Z, Kertesz Z: Antioxidant properties of di- and polyamines. Comp Biochem Physiol B 104: 475–479, 1993
Pavlovic DD, Uzunova P, Galabova T, Peneva V, Sokolova Z, Bjelakovic G, Ribarov S: Polyamines as modulators of lipoperoxidation. Gen Physiol Biophys 11: 203–211, 1992
Kafy AM, Lewis DA: Antioxidant effects of exogenous polyamines in damage of lysosomes inflicted by xanthine oxidase or stimulated polymorphonuclear leucocytes. Agents Actions 24: 145–151, 1988
Kogure K, Fukuzawa K, Kawano H, Terada H: Spermine accelerates iron-induced lipid peroxidation in mitochondria by modification of membrane surface. Free Radic Biol Med 14: 501–507, 1993
Singh A, Kumar PG, Laloraya M, Verma S, Niusarkar M: Superoxide dismutase activity regulation by spermine: a new dimension in spermine biochemistry and sperm development. Biochem Biophys Res Communs 77: 420–426, 1991
Tadolini B: The influence of polyamine-nucleic acid complexes on Fe2+ autooxidation. Mol Cell Biochem 83: 179–185, 1988
Kasai H, Nishimura S: Hydroxylation of deoxyguanosine at the C-8 position by ascorbic acid and other reducing agents. Nucleic Acids Res 12: 2137–2145, 1984
Guarnieri C, Muscari C, Stefanelli C, Giaccari A, Zini M, Di Biase S: Micellar electrokinetic capillary chromatography of 8-hydroxydeoxyguanosine and other oxidized derivatives of DNA. J Chromatogr B 656: 209–213, 1994
Pryor WA: Why is the hydroxyl radical the only radical that commonly adds to DNA? Hypothesis: it has a rare combination of high electrophilicity, high thermochemical reactivity, and a mode of production that can occur near DNA. Free Radic Biol Med 4: 219–223, 1988
Liungman M, Hanawalt PC: Efficient protection against oxidative DNA damage in chromatin. Mol Carcinog 5: 264–269, 1992
Minton KW, Tabor H, Tabor CW: Paraquat toxicity is increased inEscherichia coli defective in the synthesis of polyamines. Proc Natl Acad Sci USA 87: 2851–2855, 1990
Balasundaram D, Tabor CW, Tabor H: Oxygen toxicity in a polyamine-depletedspe2 mutant ofSaccharomyces cerevisiae. Proc Natl Acad Sci USA 90: 4693–4697, 1993
Lovaas E: Antiinflammatory and metal chelating effect of polyamines. In: C.M. Caldarera, C. Cló and M.S. Moruzzi (eds). Polyamines, Biological and Clinical Aspects, CLUEB, Bologna, Italy, 1994, pp 161–167
Shuber P: Influence of polyamines on membrane functions. Biochem J 260: 1–10, 1989
Hampel KJ, Crosson P, Lee JS: Polyamines favor DNA triplex formation at neutral pH. Biochemistry 30: 4455–4459, 1991
Rowatt E, Williams RJ: The binding of polyamines and magnesium to DNA. J Inorg Biochem 46: 87–97, 1992
Pegg AE, McCann PP: Polyamine metabolism and function. Am J Physiol 243: C212-C219, 1982
Scalabrino G, Ferioli ME: Polyamines in mammalian ageing: an oncological problem, too? A review. Mech Ageing Dev 26: 149–164, 1984
Elsayed NM: Influence of vitamine E on polyamine metabolism in ozone-exposed rat lungs. Arch Biochem Biophys 255, 392–399, 1987
Vanella A, Rapisarda A, Pinuturo R, Rizzu V: Inhibitor effect of polyamines on reduction of cytochrome C by superoxide anion. Biochem Exp Biol 16: 165–170, 1980
Bardocz S: The role of dietary polyamines. Eur J Clin Nutr 47: 683–690, 1993
Khan NA, Quemener V, Seiler N, Molinoux JP: Mechanism of spermidine uptake in cultured mammalian cells and its inhibition by some polyamine analogues. Pathobiology 58: 172–178, 1990
Nohl H, Jordan W: The mitochondrial site of superoxide formation. Biochem Biophys Res Communs 138: 533–539, 1986
Toninello A, Dalla Via L, Siliprandi D, Garlid KD: Evidence that spermine, spermidine and putrescine are transported electrophoretically in mitochondria by a specific polyamine uniporter. J Biol Chem 267: 18393–18397, 1992
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Muscari, C., Guarnieri, C., Stefanelli, C. et al. Protective effect of spermine on DNA exposed to oxidative stress. Mol Cell Biochem 144, 125–129 (1995). https://doi.org/10.1007/BF00944391
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00944391