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Formation of the spore photoproduct and other dimeric lesions between adjacent pyrimidines in UVC-irradiated dry DNA

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Abstract

Far-UV irradiation of DNA leads to the formation of several types of dimeric lesions between adjacent pyrimidine bases including cyclobutane dimers, (6-4) photoproducts and Dewar valence isomers. In the dry state, an additional specific thymine lesion, the spore photoproduct, is produced. We designed an HPLC-tandem mass spectrometry assay for the detection of the latter lesion. This technique that does not require radio-labelling of DNA allowed the simultaneous quantification of the spore photoproduct and other pyrimidine dimeric photoproducts. Using this approach, the complete distribution of bipyrimidine lesions within UVC-irradiated dry DNA was determined.

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References

  1. J. Cadet, and P. Vigny, Photochemistry of nucleic acids, in Bioorganic Photochemistry, ed. H. Morrison, New York: Wiley, 1990, vol. 1, pp. 1–272.

    CAS  Google Scholar 

  2. T. Douki, and J. Cadet, UV and nucleic acids, in Interface between Chemistry and Biochemistry (ed. H. Jornvall and P. Jollès), Birkhauser Verlag AG, Basel, 1995, pp. 173–197.

    Chapter  Google Scholar 

  3. J.-L. Ravanat, T. Douki, J. Cadet, Direct and indirect effects of UV radiation on DNA and its components, J. Photochem. Photobiol. B: Biol., 2001, 63, 88–102.

    Article  CAS  Google Scholar 

  4. J. E. Donnellan, R. B. Setlow, Thymine photoproducts but not thymine dimers found in ultraviolet-irradiated bacterial spores, Science, 1965, 149, 308–310.

    Article  CAS  Google Scholar 

  5. A. J. Varghese, 5-Thyminyl-5,6-dihydrothymine from DNA irradiated with ultraviolet light, Biochem. Biophys. Res. Comm., 1970, 38, 484–490.

    Article  CAS  Google Scholar 

  6. P. Setlow, Resistance of spores of Bacillus subtilis to ultraviolet light, Environ. Mol. Mutagen., 2001, 38, 97–104.

    Article  CAS  Google Scholar 

  7. E. Cabrera-Juarez, J. K. Setlow, Formation of a thymine photoproduct in transforming DNA by near ultraviolet irradiation, Arch. Biochem. Biophys., 1977, 475, 315–322.

    CAS  Google Scholar 

  8. C. Lindberg, G. Horneck, Thymine photoproduct formation and inactivation of intact spores of Bacillus subtilis irradiated with short wavelength UV (200-300 nm) at atmospheric pressure in vacuo, Adv. Space Res., 1992, 12, 275–279.

    Article  CAS  Google Scholar 

  9. T. A. Slieman, W. L. Nicholson, Artificial and solar UV radiation induces strand breaks and cyclobutane pyrimidine dimers in Bacillus subtilis spore DNA, Appl. Environ. Microbiol., 2000, 66, 199–205.

    Article  CAS  Google Scholar 

  10. N. Munakata, C. S. Rupert, Genetically controlled removal of “spore photoproduct” from deoxyribonucleic acid of ultraviolet-irradiated Bacillus subtilis spores, J. Bacteriol., 1972, 111, 192–198.

    Article  CAS  Google Scholar 

  11. T. S. Van Wang, C. S. Rupert, Evidence for the monomerization of spore photoproduct to two thymines by the light independent “spore repair” process in Bacillus subtilis, Photochem. Photobiol., 1977, 25, 123–127.

    Article  CAS  Google Scholar 

  12. P. Fajardo-Cavazos, C. Salazar, W. L. Nicholson, Molecular cloning and characterization of the Bacillus subtilis spore photoproduct lyase (spl) gene, which is involved in repair of UV radiation-induced DNA damage during spore germination, J. Bacteriol., 1993, 175, 1735–1744.

    Article  CAS  Google Scholar 

  13. R. A. Mehl, T. P. Begley, Mechanistic studies on the repair of a novel DNA photolesion: the spore photoproduct, Org. Lett., 1999, 1, 1065–1066.

    Article  CAS  Google Scholar 

  14. R. Rebeil, W. L. Nicholson, The subunit structure and catalytic mechanism of Bacillus subtilis DNA repair enzyme spore photoproduct lyase, Proc. Natl. Acad. Sci. USA, 2001, 98, 9038–9043.

    Article  CAS  Google Scholar 

  15. J. Cheek, J. B. Broderick, Direct H Atom abstraction from spore photoproduct C-6 initiates DNA repair in the reaction catalyzed by spore photoproduct lyase: Evidence for a reversibly generated adenosyl radical intermediate, J. Am. Chem. Soc., 2002, 124, 2860–2861.

    Article  CAS  Google Scholar 

  16. D. L. Mitchell, J. M. Clarkson, Use of synthetic polynucleotides to characterize an antiserum made against UV-irradiated DNA, Photochem. Photobiol., 1984, 40, 743–748.

    Article  CAS  Google Scholar 

  17. T. Mori, M. Nakane, T. Hattori, T. Matsunaga, M. Ihara, O. Nikaido, Simultaneous establishment of monoclonal antibodies specific for either cyclobutane pyrimidine dimer or (6-4) photoproduct from the same mouse immunized with ultraviolet-irradiated DNA, Photochem. Photobiol., 1991, 54, 225–232.

    Article  CAS  Google Scholar 

  18. T. Matsunaga, Y. Hatakeyama, M. Ohta, T. Mori, O. Nikaido, Establishment and characterization of a monoclonal antibody recognizing the Dewar valence isomer of (6-4) photoproducts, Photochem. Photobiol., 1993, 57, 934–940.

    Article  CAS  Google Scholar 

  19. P. W. Doetsch, G. L. Chan, W. A. Haseltine, T4 DNA polymerase (3’-5’) exonuclease, an enzyme for the detection and quantification of stable DNA lesions: the ultraviolet example, Nucleic Acids Res., 1985, 13, 3285–3302.

    Article  CAS  Google Scholar 

  20. E. Sage, E. Cramb, B. W. Glickman, The distribution of UV damage in the lacI gene of Escherichia coli: correlation with mutation spectrum, Mutat. Res., 1992, 269, 285–299.

    Article  CAS  Google Scholar 

  21. G. P. Pfeifer, R. Drouin, G. P. Holmquist, Detection of DNA adducts at the DNA sequence level by ligation-mediated PCR, Mutat. Res., 1993, 288, 39–46.

    Article  CAS  Google Scholar 

  22. M. H. Patrick, D. M. Gray, Independence of photoproduct formation on DNA conformation, Photochem. Photobiol., 1976, 24, 507–513.

    Article  CAS  Google Scholar 

  23. H. J. Niggli, P. A. Cerutti, Cyclobutane-type pyrimidine dimer formation and excision in human skin fibroblasts after irradiation with 313 nm ultraviolet light, Biochemistry, 1983, 22, 1390–1395.

    Article  CAS  Google Scholar 

  24. J. Cadet, N. E. Gentner, B. Rozga, M. C. Paterson, Rapid quantitation of ultraviolet induced thymine-containing dimers in human cell DNA by reverse-phase high performance liquid chromatography, J. Chromatogr., 1983, 280, 99–108.

    Article  CAS  Google Scholar 

  25. T. Douki, L. Voituriez, J. Cadet, Measurement of pyrimidine (6-4) pyrimidone photoproducts in DNA by a mild acidic hydrolysis-HPLC fluorescence assay, Chem. Res. Toxicol., 1995, 8, 244–253.

    Article  CAS  Google Scholar 

  26. Y. Sun, K. Palasingam, W. L. Nicholson, High-pressure liquid chromatography assay for quantitatively monitoring spore photoproduct repair mediated by spore photoproduct lyase during germination of UV-irradiated Bacillus subtilis spores, Anal. Biochem., 1994, 221, 61–65.

    Article  CAS  Google Scholar 

  27. T. Douki, M. Court, S. Sauvaigo, F. Odin, J. Cadet, Formation of the main UV-induced thymine dimeric lesions within isolated and cellular DNA as measured by HPLC-MS/MS, J. Biol. Chem., 2000, 275, 11678–11685.

    Article  CAS  Google Scholar 

  28. T. Douki, J. Cadet, Individual determination of the yield of the main-UV induced dimeric pyrimidine photoproducts in DNA suggests a high mutagenicity of CC photolesions, Biochemistry, 2001, 40, 2495–2501.

    Article  CAS  Google Scholar 

  29. T. Douki, M. Court, J. Cadet, Electrospray-mass spectrometry characterization and detection of far-UV induced thymine photoproducts, J. Photochem. Photobiol. B: Biol., 2000, 54, 145–154.

    Article  CAS  Google Scholar 

  30. M. Liuzzi, M. Weinfeld, M. C. Paterson, Enzymatic analysis of isomeric trithymidilates containing ultraviolet light-induced cyclobutane pyrimidine dimers I Nuclease P1 mediated hydrolysis of the intradimer phosphodiester linkage, J. Biol. Chem., 1989, 264, 6355–6363.

    Article  CAS  Google Scholar 

  31. T. Douki, T. Zalizniak, J. Cadet, Far-UV-induced dimeric photoproducts in short oligonucleotides: Sequence effects, Photochem. Photobiol., 1997, 66, 171–179.

    Article  CAS  Google Scholar 

  32. T. Douki, D. Angelov, J. Cadet, UV Laser photolysis of DNA: effect of duplex stability on charge-transfer efficiency, J. Am. Chem. Soc., 2001, 123, 11360–11366.

    Article  CAS  Google Scholar 

  33. R. O. Rahn, J. L. Hosszu, Influence of relative humidity on the photochemistry of DNA films, Biochim. Biophys. Acta, 1969, 190, 126–131.

    Article  CAS  Google Scholar 

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Correspondence to Thierry Douki.

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Douki, T., Cadet, J. Formation of the spore photoproduct and other dimeric lesions between adjacent pyrimidines in UVC-irradiated dry DNA. Photochem Photobiol Sci 2, 433–436 (2003). https://doi.org/10.1039/b300173c

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  • DOI: https://doi.org/10.1039/b300173c

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