Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1987 Oct;84(20):7070–7074. doi: 10.1073/pnas.84.20.7070

3'-Formyl phosphate-ended DNA: high-energy intermediate in antibiotic-induced DNA sugar damage.

D H Chin 1, L S Kappen 1, I H Goldberg 1
PMCID: PMC299231  PMID: 2959956

Abstract

Under anaerobic conditions where the nitroaromatic radiation-sensitizer misonidazole substitutes for dioxygen, DNA strand breakage (gaps with phosphate residues at each end) by the nonprotein chromophore of the antitumor antibiotic neocarzinostatin (NCS-Chrom) is associated with the generation of a reactive form of formate from the C-5' of deoxyribose of thymidylate residues. Such lesions account for a minority (10-15%) of the strand breakage found in the aerobic reaction without misonidazole. Amino-containing nucleophiles such as tris(hydroxymethyl)aminomethane (Tris) and hydroxylamine act as acceptors for the activated formate. The amount of [3H]formyl hydroxamate produced from DNA labeled with [5'-3H]thymidine is comparable to the spontaneously released thymine. During the course of the reaction, misonidazole undergoes a DNA-dependent reduction and subsequent conjugation with glutathione used to activate NCS-Chrom. From these and earlier results, we propose a possible mechanism in which the carbon-centered radical formed at C-5' by hydrogen atom abstraction by thiol-activated NCS-Chrom reacts anaerobically with misonidazole to form a nitroxyl-radical-adduct intermediate, which fragments to produce an oxy radical at C-5'. beta-Fragmentation results in cleavage between C-5' and C-4' with the generation of 3'-formyl phosphate-ended DNA, a high-energy form of formate, which spontaneously hydrolyzes, releasing formate and creating a 3'-phosphate end, or transfers the formyl moiety to available nucleophiles. A similar mechanism, involving dioxygen addition, is probably responsible for the 10-15% DNA gap formation in the aerobic reaction.

Full text

PDF
7071

Selected References

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

  1. Chin D. H., Carr S. A., Goldberg I. H. Incorporation of 18O2 into thymidine 5'-aldehyde in neocarzinostatin chromophore-damaged DNA. J Biol Chem. 1984 Aug 25;259(16):9975–9978. [PubMed] [Google Scholar]
  2. Goldberg I. H. Novel types of DNA-sugar damage in neocarzinostatin cytotoxicity and mutagenesis. Basic Life Sci. 1986;38:231–244. doi: 10.1007/978-1-4615-9462-8_24. [DOI] [PubMed] [Google Scholar]
  3. Hatayama T., Goldberg I. H. Deoxyribonucleic acid sugar damage in the action of neocarzinostatin. Biochemistry. 1980 Dec 9;19(25):5890–5898. doi: 10.1021/bi00566a035. [DOI] [PubMed] [Google Scholar]
  4. Kappen L. S., Ellenberger T. E., Goldberg I. H. Mechanism and base specificity of DNA Breakage in intact cells by neocarzinostatin. Biochemistry. 1987 Jan 27;26(2):384–390. doi: 10.1021/bi00376a008. [DOI] [PubMed] [Google Scholar]
  5. Kappen L. S., Goldberg I. H. Activation of neocarzinostatin chromophore and formation of nascent DNA damage do not require molecular oxygen. Nucleic Acids Res. 1985 Mar 11;13(5):1637–1648. doi: 10.1093/nar/13.5.1637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kappen L. S., Goldberg I. H. Deoxyribonucleic acid damage by neocarzinostatin chromophore: strand breaks generated by selective oxidation of C-5' of deoxyribose. Biochemistry. 1983 Oct 11;22(21):4872–4878. doi: 10.1021/bi00290a002. [DOI] [PubMed] [Google Scholar]
  7. Kappen L. S., Goldberg I. H. Nitroaromatic radiation sensitizers substitute for oxygen in neocarzinostatin-induced DNA damage. Proc Natl Acad Sci U S A. 1984 Jun;81(11):3312–3316. doi: 10.1073/pnas.81.11.3312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Nishimoto S., Ide H., Wada T., Kagiya T. Radiation-induced hydroxylation of thymine promoted by electron-affinic compounds. Int J Radiat Biol Relat Stud Phys Chem Med. 1983 Dec;44(6):585–600. doi: 10.1080/09553008314551651. [DOI] [PubMed] [Google Scholar]
  9. Poon R., Beerman T. A., Goldberg I. H. Characterization of DNA strand breakage in vitro by the antitumor protein neocarzinostatin. Biochemistry. 1977 Feb 8;16(3):486–493. doi: 10.1021/bi00622a023. [DOI] [PubMed] [Google Scholar]
  10. Povirk L. F., Goldberg I. H. Endonuclease-resistant apyrimidinic sites formed by neocarzinostatin at cytosine residues in DNA: evidence for a possible role in mutagenesis. Proc Natl Acad Sci U S A. 1985 May;82(10):3182–3186. doi: 10.1073/pnas.82.10.3182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Varghese A. J. Glutathione conjugates of misonidazole. Biochem Biophys Res Commun. 1983 May 16;112(3):1013–1020. doi: 10.1016/0006-291x(83)91719-9. [DOI] [PubMed] [Google Scholar]
  12. Varghese A. J., Whitmore G. F. Cellular and chemical reduction products of misonidazole. Chem Biol Interact. 1981 Aug;36(2):141–151. doi: 10.1016/0009-2797(81)90016-8. [DOI] [PubMed] [Google Scholar]
  13. WUNDERLY C. A trough for paper chromatography consisting of segments. Nature. 1954 Feb 6;173(4397):267–268. doi: 10.1038/173267b0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES