PMID: 2495939Jan 1, 1989Paper

Rapid detection of DNA-interstrand and DNA-protein cross-links in mammalian cells by gravity-flow alkaline elution

Environmental and Molecular Mutagenesis
J R Hincks, R A Coulombe

Abstract

Alkaline elution is a sensitive and commonly used technique to detect cellular DNA damage in the form of DNA strand breaks and DNA cross-links. Conventional alkaline elution procedures have extensive equipment requirements and are tedious to perform. Our laboratory recently presented a rapid, simplified, and sensitive modification of the alkaline elution technique to detect carcinogen-induced DNA strand breaks. In the present study, we have further modified this technique to enable the rapid characterization of chemically induced DNA-interstrand and DNA-protein associated cross-links in cultured epithelial cells. Cells were exposed to three known DNA cross-linking agents, nitrogen mustard (HN2), mitomycin C (MMC), or ultraviolet irradiation (UV). One hour exposures of HN2 at 0.25, 1.0, and 4.0 microM or of MMC at 20, 40, and 60 microM produced a dose-dependent induction of total DNA cross-links by these agents. Digestion with proteinase K revealed that HN2 and MMC induced both DNA-protein cross-links and DNA-interstrand cross-links. Ultraviolet irradiation induced both DNA cross-links and DNA strand breaks, the latter of which were either protein and nonprotein associated. The results demonstrate that gravity-flow alkaline elut...Continue Reading

References

Jan 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·A J FornaceH E Kann
Oct 19, 1976·Biochemistry·K W KohnC A Friedman
Aug 16, 1977·Biochimica Et Biophysica Acta·A J Fornace, J B Little
Sep 20, 1976·Biochemical and Biophysical Research Communications·J A SwenbergP R Harbach
Jun 30, 1986·Biochemical and Biophysical Research Communications·J R Hincks, R A Coulombe
Jan 1, 1987·Toxicology and Applied Pharmacology·N C MargarettenR A Coulombe
Apr 14, 1967·Journal of Molecular Biology·P D Lawley, P Brookes
Aug 1, 1963·Proceedings of the National Academy of Sciences of the United States of America·V N IYER, W SZYBALSKI

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Citations

Jan 1, 1991·Pharmacology & Therapeutics·K W Kohn
Oct 11, 1991·Toxicology and Applied Pharmacology·J R HincksR A Coulombe
Jan 1, 1996·Environmental and Molecular Mutagenesis·S Pfuhler, H U Wolf
May 19, 2021·Archives of Toxicology·Yisheng HeGe Lin

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