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. 1992 Aug 1;89(15):6866–6870. doi: 10.1073/pnas.89.15.6866

Mutations induced by methylene blue plus light in single-stranded M13mp2.

T J McBride 1, J E Schneider 1, R A Floyd 1, L A Loeb 1
PMCID: PMC49605  PMID: 1495976

Abstract

Reactive oxygen species are generated by a variety of cellular processes. These endogenously generated, reactive intermediates produce a multiplicity of DNA alterations and mutations and have been implicated in the pathogenesis of several human diseases. We report here that treatment of single-stranded M13mp2 bacteriophage DNA with methylene blue and white light generates increased levels of 8-hydroxydeoxyguanosine and that mutagenesis is both highly specific and dependent on the SOS response. Lesions produced block the progression of DNA synthesis one base preceding template guanines. In SOS-induced Escherichia coli, 97% of all methylene blue-induced mutations in the lacZ alpha gene of M13mp2 DNA are single-base substitutions opposite template guanines. The most frequent mutations are G----C transversions. The G----T transversions expected from the presence of 8-hydroxydeoxyguanosine in the template strand occur, but at a lower frequency. Sequence data together with SOS dependency and the presence of replication blockage demonstrate that while 8-hydroxydeoxyguanosine may serve as an important marker to monitor oxygen-induced DNA damage in humans, it does not account for either the observed blockage to replication or the mutagenesis by methylene blue plus light in SOS-induced E. coli. Instead, an as yet unidentified lesion generated by active oxygen species is a more potent mutagenic event.

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Selected References

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

  1. Ames B. N. Endogenous DNA damage as related to cancer and aging. Mutat Res. 1989 Sep;214(1):41–46. doi: 10.1016/0027-5107(89)90196-6. [DOI] [PubMed] [Google Scholar]
  2. Boiteux S., Laval J. Imidazole open ring 7-methylguanine: an inhibitor of DNA synthesis. Biochem Biophys Res Commun. 1983 Jan 27;110(2):552–558. doi: 10.1016/0006-291x(83)91185-3. [DOI] [PubMed] [Google Scholar]
  3. Boiteux S., O'Connor T. R., Lederer F., Gouyette A., Laval J. Homogeneous Escherichia coli FPG protein. A DNA glycosylase which excises imidazole ring-opened purines and nicks DNA at apurinic/apyrimidinic sites. J Biol Chem. 1990 Mar 5;265(7):3916–3922. [PubMed] [Google Scholar]
  4. Cadet J., Berger M. Radiation-induced decomposition of the purine bases within DNA and related model compounds. Int J Radiat Biol Relat Stud Phys Chem Med. 1985 Feb;47(2):127–143. doi: 10.1080/09553008514550201. [DOI] [PubMed] [Google Scholar]
  5. Cerutti P. A. Prooxidant states and tumor promotion. Science. 1985 Jan 25;227(4685):375–381. doi: 10.1126/science.2981433. [DOI] [PubMed] [Google Scholar]
  6. Cheng K. C., Cahill D. S., Kasai H., Nishimura S., Loeb L. A. 8-Hydroxyguanine, an abundant form of oxidative DNA damage, causes G----T and A----C substitutions. J Biol Chem. 1992 Jan 5;267(1):166–172. [PubMed] [Google Scholar]
  7. Chung M. H., Kim H. S., Ohtsuka E., Kasai H., Yamamoto F., Nishimura S. An endonuclease activity in human polymorphonuclear neutrophils that removes 8-hydroxyguanine residues from DNA+. Biochem Biophys Res Commun. 1991 Aug 15;178(3):1472–1478. doi: 10.1016/0006-291x(91)91059-l. [DOI] [PubMed] [Google Scholar]
  8. Czeczot H., Tudek B., Lambert B., Laval J., Boiteux S. Escherichia coli Fpg protein and UvrABC endonuclease repair DNA damage induced by methylene blue plus visible light in vivo and in vitro. J Bacteriol. 1991 Jun;173(11):3419–3424. doi: 10.1128/jb.173.11.3419-3424.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Das S. K., Benditt E. P., Loeb L. A. Rapid changes in deoxynucleoside triphosphate pools in mammalian cells treated with mutagens. Biochem Biophys Res Commun. 1983 Jul 29;114(2):458–464. doi: 10.1016/0006-291x(83)90802-1. [DOI] [PubMed] [Google Scholar]
  10. Decuyper-Debergh D., Piette J., Van de Vorst A. Singlet oxygen-induced mutations in M13 lacZ phage DNA. EMBO J. 1987 Oct;6(10):3155–3161. doi: 10.1002/j.1460-2075.1987.tb02626.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Del Sal G., Manfioletti G., Schneider C. The CTAB-DNA precipitation method: a common mini-scale preparation of template DNA from phagemids, phages or plasmids suitable for sequencing. Biotechniques. 1989 May;7(5):514–520. [PubMed] [Google Scholar]
  12. Devasagayam T. P., Steenken S., Obendorf M. S., Schulz W. A., Sies H. Formation of 8-hydroxy(deoxy)guanosine and generation of strand breaks at guanine residues in DNA by singlet oxygen. Biochemistry. 1991 Jun 25;30(25):6283–6289. doi: 10.1021/bi00239a029. [DOI] [PubMed] [Google Scholar]
  13. Dizdaroglu M., Bergtold D. S. Characterization of free radical-induced base damage in DNA at biologically relevant levels. Anal Biochem. 1986 Jul;156(1):182–188. doi: 10.1016/0003-2697(86)90171-5. [DOI] [PubMed] [Google Scholar]
  14. Epe B., Hegler J., Wild D. Singlet oxygen as an ultimately reactive species in Salmonella typhimurium DNA damage induced by methylene blue/visible light. Carcinogenesis. 1989 Nov;10(11):2019–2024. doi: 10.1093/carcin/10.11.2019. [DOI] [PubMed] [Google Scholar]
  15. Floyd R. A. Role of oxygen free radicals in carcinogenesis and brain ischemia. FASEB J. 1990 Jun;4(9):2587–2597. [PubMed] [Google Scholar]
  16. Floyd R. A., Watson J. J., Wong P. K., Altmiller D. H., Rickard R. C. Hydroxyl free radical adduct of deoxyguanosine: sensitive detection and mechanisms of formation. Free Radic Res Commun. 1986;1(3):163–172. doi: 10.3109/10715768609083148. [DOI] [PubMed] [Google Scholar]
  17. Floyd R. A., West M. S., Eneff K. L., Schneider J. E. Methylene blue plus light mediates 8-hydroxyguanine formation in DNA. Arch Biochem Biophys. 1989 Aug 15;273(1):106–111. doi: 10.1016/0003-9861(89)90167-7. [DOI] [PubMed] [Google Scholar]
  18. Fraga C. G., Shigenaga M. K., Park J. W., Degan P., Ames B. N. Oxidative damage to DNA during aging: 8-hydroxy-2'-deoxyguanosine in rat organ DNA and urine. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4533–4537. doi: 10.1073/pnas.87.12.4533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Harris C. C. Chemical and physical carcinogenesis: advances and perspectives for the 1990s. Cancer Res. 1991 Sep 15;51(18 Suppl):5023s–5044s. [PubMed] [Google Scholar]
  20. Kasai H., Crain P. F., Kuchino Y., Nishimura S., Ootsuyama A., Tanooka H. Formation of 8-hydroxyguanine moiety in cellular DNA by agents producing oxygen radicals and evidence for its repair. Carcinogenesis. 1986 Nov;7(11):1849–1851. doi: 10.1093/carcin/7.11.1849. [DOI] [PubMed] [Google Scholar]
  21. Koffel-Schwartz N., Maenhaut-Michel G., Fuchs R. P. Specific strand loss in N-2-acetylaminofluorene-modified DNA. J Mol Biol. 1987 Feb 20;193(4):651–659. doi: 10.1016/0022-2836(87)90348-2. [DOI] [PubMed] [Google Scholar]
  22. Kunkel T. A. Mutational specificity of depurination. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1494–1498. doi: 10.1073/pnas.81.5.1494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kunkel T. A. The mutational specificity of DNA polymerase-beta during in vitro DNA synthesis. Production of frameshift, base substitution, and deletion mutations. J Biol Chem. 1985 May 10;260(9):5787–5796. [PubMed] [Google Scholar]
  24. McBride T. J., Preston B. D., Loeb L. A. Mutagenic spectrum resulting from DNA damage by oxygen radicals. Biochemistry. 1991 Jan 8;30(1):207–213. doi: 10.1021/bi00215a030. [DOI] [PubMed] [Google Scholar]
  25. Moriya M., Ou C., Bodepudi V., Johnson F., Takeshita M., Grollman A. P. Site-specific mutagenesis using a gapped duplex vector: a study of translesion synthesis past 8-oxodeoxyguanosine in E. coli. Mutat Res. 1991 May;254(3):281–288. doi: 10.1016/0921-8777(91)90067-y. [DOI] [PubMed] [Google Scholar]
  26. Müller E., Boiteux S., Cunningham R. P., Epe B. Enzymatic recognition of DNA modifications induced by singlet oxygen and photosensitizers. Nucleic Acids Res. 1990 Oct 25;18(20):5969–5973. doi: 10.1093/nar/18.20.5969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Naqui A., Chance B., Cadenas E. Reactive oxygen intermediates in biochemistry. Annu Rev Biochem. 1986;55:137–166. doi: 10.1146/annurev.bi.55.070186.001033. [DOI] [PubMed] [Google Scholar]
  28. Park J. W., Cundy K. C., Ames B. N. Detection of DNA adducts by high-performance liquid chromatography with electrochemical detection. Carcinogenesis. 1989 May;10(5):827–832. doi: 10.1093/carcin/10.5.827. [DOI] [PubMed] [Google Scholar]
  29. Piette J. Biological consequences associated with DNA oxidation mediated by singlet oxygen. J Photochem Photobiol B. 1991 Dec;11(3-4):241–260. doi: 10.1016/1011-1344(91)80030-l. [DOI] [PubMed] [Google Scholar]
  30. Piette J., Calberg-Bacq C. M., Lopez M., van de Vorst A. Terminations of DNA synthesis on 'proflavine and light'-treated phi X174 single-stranded DNA. Biochim Biophys Acta. 1984 Apr 5;781(3):257–264. doi: 10.1016/0167-4781(84)90091-5. [DOI] [PubMed] [Google Scholar]
  31. Piette J., Merville-Louis M. P., Decuyper J. Damages induced in nucleic acids by photosensitization. Photochem Photobiol. 1986 Dec;44(6):793–802. doi: 10.1111/j.1751-1097.1986.tb05539.x. [DOI] [PubMed] [Google Scholar]
  32. SIMON M. I., GROSSMAN L., VANVUNAKIS H. PHOTOSENSITIZED REACTION OF POLYRIBONUCLEOTIDES. I. EFFECTS ON THEIR SUSCEPTIBILITY TO ENZYME DIGESTION AND THEIR ABILITY TO ACT AS SYNTHETIC MESSENGERS. J Mol Biol. 1965 May;12:50–59. doi: 10.1016/s0022-2836(65)80281-9. [DOI] [PubMed] [Google Scholar]
  33. SIMON M. I., VAN VUNAKIS H. The photodynamic reaction of methylene blue with deoxyribonucleic acid. J Mol Biol. 1962 Jun;4:488–499. doi: 10.1016/s0022-2836(62)80104-1. [DOI] [PubMed] [Google Scholar]
  34. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schaaper R. M., Kunkel T. A., Loeb L. A. Infidelity of DNA synthesis associated with bypass of apurinic sites. Proc Natl Acad Sci U S A. 1983 Jan;80(2):487–491. doi: 10.1073/pnas.80.2.487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Schneider J. E., Price S., Maidt L., Gutteridge J. M., Floyd R. A. Methylene blue plus light mediates 8-hydroxy 2'-deoxyguanosine formation in DNA preferentially over strand breakage. Nucleic Acids Res. 1990 Feb 11;18(3):631–635. doi: 10.1093/nar/18.3.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Shigenaga M. K., Gimeno C. J., Ames B. N. Urinary 8-hydroxy-2'-deoxyguanosine as a biological marker of in vivo oxidative DNA damage. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9697–9701. doi: 10.1073/pnas.86.24.9697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Spahr R. C., Salsburey D. J., Krissberg A., Prin W. Intraamniotic injection of methylene blue leading to methemoglobinemia in one of twins. Int J Gynaecol Obstet. 1980 Mar-Apr;17(5):477–478. doi: 10.1002/j.1879-3479.1980.tb00192.x. [DOI] [PubMed] [Google Scholar]
  39. Tchou J., Kasai H., Shibutani S., Chung M. H., Laval J., Grollman A. P., Nishimura S. 8-oxoguanine (8-hydroxyguanine) DNA glycosylase and its substrate specificity. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4690–4694. doi: 10.1073/pnas.88.11.4690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tkeshelashvili L. K., McBride T., Spence K., Loeb L. A. Mutation spectrum of copper-induced DNA damage. J Biol Chem. 1991 Apr 5;266(10):6401–6406. [PubMed] [Google Scholar]
  41. Waskell L. A., Sastry K. S., Gordon M. P. Studies on the photosensitized breakdown of guanosine by methylene blue. Biochim Biophys Acta. 1966 Oct 24;129(1):49–53. doi: 10.1016/0005-2787(66)90007-4. [DOI] [PubMed] [Google Scholar]
  42. Wood M. L., Dizdaroglu M., Gajewski E., Essigmann J. M. Mechanistic studies of ionizing radiation and oxidative mutagenesis: genetic effects of a single 8-hydroxyguanine (7-hydro-8-oxoguanine) residue inserted at a unique site in a viral genome. Biochemistry. 1990 Jul 31;29(30):7024–7032. doi: 10.1021/bi00482a011. [DOI] [PubMed] [Google Scholar]

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