Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1976 Feb;3(2):315–323. doi: 10.1093/nar/3.2.315

Studies on the mechanism of DNA cleavage by ethidium.

I S Deniss, A R Morgan
PMCID: PMC342903  PMID: 1257050

Abstract

Ethidium causes the cleavage of DNA via a light and oxygen dependent process. Using covalently closed circular DNA as a substrate, the saturation kinetics and the dependence on superhelical density of the cleavage indicate that intercalated ethidium is mainly responsible for nicking DNA. Superoxide dismutase has little effect on the reaction and catalase none. Lowering the pH inhibited the reaction. The reaction mechanism and its use in determining superhelical densities of covalently closed circular DNA's are discussed.

Full text

PDF
315

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Balny C., Douzou P. Production of superoxide ions by photosensitization of dyes. Biochem Biophys Res Commun. 1974 Jan 23;56(2):386–391. doi: 10.1016/0006-291x(74)90854-7. [DOI] [PubMed] [Google Scholar]
  2. Bauer W., Vinograd J. The interaction of closed circular DNA with intercalative dyes. I. The superhelix density of SV40 DNA in the presence and absence of dye. J Mol Biol. 1968 Apr 14;33(1):141–171. doi: 10.1016/0022-2836(68)90286-6. [DOI] [PubMed] [Google Scholar]
  3. Freifelder D. Electron microscopic study of the ethidium bromide-DNA complex. J Mol Biol. 1971 Sep 14;60(2):401–403. doi: 10.1016/0022-2836(71)90303-2. [DOI] [PubMed] [Google Scholar]
  4. Kline B. C., Helinski D. R. F 1 sex factor of Escherichia coli. Size and purification in the form of a strand-specific relaxation complex of supercoiled deoxyribonucleic acid and protein. Biochemistry. 1971 Dec 21;10(26):4975–4980. doi: 10.1021/bi00802a022. [DOI] [PubMed] [Google Scholar]
  5. LePecq J. B., Paoletti C. A fluorescent complex between ethidium bromide and nucleic acids. Physical-chemical characterization. J Mol Biol. 1967 Jul 14;27(1):87–106. doi: 10.1016/0022-2836(67)90353-1. [DOI] [PubMed] [Google Scholar]
  6. McCord J. M. Free radicals and inflammation: protection of synovial fluid by superoxide dismutase. Science. 1974 Aug 9;185(4150):529–531. doi: 10.1126/science.185.4150.529. [DOI] [PubMed] [Google Scholar]
  7. McCord J. M., Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem. 1969 Nov 25;244(22):6049–6055. [PubMed] [Google Scholar]
  8. McCord J. M., Fridovich I. The utility of superoxide dismutase in studying free radical reactions. I. Radicals generated by the interaction of sulfite, dimethyl sulfoxide, and oxygen. J Biol Chem. 1969 Nov 25;244(22):6056–6063. [PubMed] [Google Scholar]
  9. Morgan A. R., Pulleyblank D. E. Native and denatured DNA, cross-linked and palindromic DNA and circular covalently-closed DNA analysed by a sensitive fluorometric procedure. Biochem Biophys Res Commun. 1974 Nov 27;61(2):396–403. doi: 10.1016/0006-291x(74)90970-x. [DOI] [PubMed] [Google Scholar]
  10. Pulleyblank D. E., Morgan A. R. The sense of naturally occurring superhelices and the unwinding angle of intercalated ethidium. J Mol Biol. 1975 Jan 5;91(1):1–13. doi: 10.1016/0022-2836(75)90368-x. [DOI] [PubMed] [Google Scholar]
  11. Wang J. C. Interactions between twisted DNAs and enzymes: the effects of superhelical turns. J Mol Biol. 1974 Aug 25;87(4):797–816. doi: 10.1016/0022-2836(74)90085-0. [DOI] [PubMed] [Google Scholar]
  12. Wang J. C. The degree of unwinding of the DNA helix by ethidium. I. Titration of twisted PM2 DNA molecules in alkaline cesium chloride density gradients. J Mol Biol. 1974 Nov 15;89(4):783–801. doi: 10.1016/0022-2836(74)90053-9. [DOI] [PubMed] [Google Scholar]
  13. Waring M. J. Drugs which affect the structure and function of DNA. Nature. 1968 Sep 28;219(5161):1320–1325. doi: 10.1038/2191320a0. [DOI] [PubMed] [Google Scholar]
  14. Worcel A., Burgi E. On the structure of the folded chromosome of Escherichia coli. J Mol Biol. 1972 Nov 14;71(2):127–147. doi: 10.1016/0022-2836(72)90342-7. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES