Skip to main content
NCBI 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
. 1982 Oct;79(19):5971–5975. doi: 10.1073/pnas.79.19.5971

SOS chromotest, a direct assay of induction of an SOS function in Escherichia coli K-12 to measure genotoxicity.

P Quillardet, O Huisman, R D'Ari, M Hofnung
PMCID: PMC347033  PMID: 6821127

Abstract

We present and evaluate the SOS chromotest, a bacterial test for detecting DNA-damaging agents. It is a colorimetric assay based on the induction by these agents of the SOS function sfiA, whose level of expression is monitored by means of a sfiA::lacZ operon fusion. The response is rapid (a few hours), and does not require survival of the tester strain. Dose-response curves for various chemicals include a linear region. The slope of this region is taken as a measure of the SOS inducing potency. Comparison for a number of substances of known genotoxicity of the SOS inducing potency determined in the SOS chromotest with the mutagenic potency determined in the Salmonella assay (mutatest) revealed a striking quantitative correlation over more than 7 orders of magnitude. The sensitivity of the SOS chromotest (lowest amount detected) is equal to that of the mutatest and generally 4-40 times higher than that of a phage induction assay (inductest). From a practical point of view our observations contribute to the validation of the SOS chromotest as a test for detecting genotoxins and in particular genotoxic carcinogens. From a theoretical standpoint the results suggest that mutagenic potency measured in the mutatest reflects the level of induction of an SOS function and that most genotoxins are inducers of the SOS response in bacteria.

Full text

PDF
5971

Selected References

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

  1. Ames B. N., Durston W. E., Yamasaki E., Lee F. D. Carcinogens are mutagens: a simple test system combining liver homogenates for activation and bacteria for detection. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2281–2285. doi: 10.1073/pnas.70.8.2281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ames B. N., Lee F. D., Durston W. E. An improved bacterial test system for the detection and classification of mutagens and carcinogens. Proc Natl Acad Sci U S A. 1973 Mar;70(3):782–786. doi: 10.1073/pnas.70.3.782. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ames B. N., Mccann J., Yamasaki E. Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test. Mutat Res. 1975 Dec;31(6):347–364. doi: 10.1016/0165-1161(75)90046-1. [DOI] [PubMed] [Google Scholar]
  4. Bagg A., Kenyon C. J., Walker G. C. Inducibility of a gene product required for UV and chemical mutagenesis in Escherichia coli. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5749–5753. doi: 10.1073/pnas.78.9.5749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Barrett J. C. Induction of gene mutation in and cell transformation of mammalian cells by modified purines: 2-aminopurine and 6-N-hydroxylaminopurine. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5685–5689. doi: 10.1073/pnas.78.9.5685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bartsch H., Malaveille C., Camus A. M., Martel-Planche G., Brun G., Hautefeuille A., Sabadie N., Barbin A., Kuroki T., Drevon C. Validation and comparative studies on 180 chemicals with S. typhimurium strains and V79 Chinese hamster cells in the presence of various metabolizing systems. Mutat Res. 1980 Jul;76(1):1–50. doi: 10.1016/0165-1110(80)90002-0. [DOI] [PubMed] [Google Scholar]
  7. Boyle J. M., Setlow R. B. Correlations between host-cell reactivation, ultraviolet reactivation and pyrimidine dimer excision in the DNA of bacteriophage lambda. J Mol Biol. 1970 Jul 14;51(1):131–144. doi: 10.1016/0022-2836(70)90275-5. [DOI] [PubMed] [Google Scholar]
  8. Cairns J. The origin of human cancers. Nature. 1981 Jan 29;289(5796):353–357. doi: 10.1038/289353a0. [DOI] [PubMed] [Google Scholar]
  9. Devoret R. Inducible error-prone repair and induction of prophage lambda in Escherichia coli. Prog Nucleic Acid Res Mol Biol. 1981;26:251–263. doi: 10.1016/s0079-6603(08)60410-9. [DOI] [PubMed] [Google Scholar]
  10. Elespuru R. K., Yarmolinsky M. B. A colorimetric assay of lysogenic induction designed for screening potential carcinogenic and carcinostatic agents. Environ Mutagen. 1979;1(1):65–78. doi: 10.1002/em.2860010113. [DOI] [PubMed] [Google Scholar]
  11. Hanawalt P. C., Cooper P. K., Ganesan A. K., Smith C. A. DNA repair in bacteria and mammalian cells. Annu Rev Biochem. 1979;48:783–836. doi: 10.1146/annurev.bi.48.070179.004031. [DOI] [PubMed] [Google Scholar]
  12. Ho Y. L., Ho S. K. Screening of carcinogens with the prophage lambda cIts857 induction test. Cancer Res. 1981 Feb;41(2):532–536. [PubMed] [Google Scholar]
  13. Hofnung M., Weill N. Inductest and spermatest in genetic toxicology testing. Arch Toxicol. 1980 Nov;46(1-2):159–169. doi: 10.1007/BF00361254. [DOI] [PubMed] [Google Scholar]
  14. Hollstein M., McCann J., Angelosanto F. A., Nichols W. W. Short-term tests for carcinogens and mutagens. Mutat Res. 1979 Sep;65(3):133–226. doi: 10.1016/0165-1110(79)90014-9. [DOI] [PubMed] [Google Scholar]
  15. Horii T., Ogawa T., Nakatani T., Hase T., Matsubara H., Ogawa H. Regulation of SOS functions: purification of E. coli LexA protein and determination of its specific site cleaved by the RecA protein. Cell. 1981 Dec;27(3 Pt 2):515–522. doi: 10.1016/0092-8674(81)90393-7. [DOI] [PubMed] [Google Scholar]
  16. Huisman O., D'Ari R. An inducible DNA replication-cell division coupling mechanism in E. coli. Nature. 1981 Apr 30;290(5809):797–799. doi: 10.1038/290797a0. [DOI] [PubMed] [Google Scholar]
  17. ISHIDA N., MIYAZAKI K., KUMAGAI K., RIKIMARU M. NEOCARZINOSTATIN, AN ANTITUMOR ANTIBIOTIC OF HIGH MOLECULAR WEIGHT. ISOLATION, PHYSIOCHEMICAL PROPERTIES AND BIOLOGICAL ACTIVITIES. J Antibiot (Tokyo) 1965 Mar;18:68–76. [PubMed] [Google Scholar]
  18. Kato T., Shinoura Y. Isolation and characterization of mutants of Escherichia coli deficient in induction of mutations by ultraviolet light. Mol Gen Genet. 1977 Nov 14;156(2):121–131. doi: 10.1007/BF00283484. [DOI] [PubMed] [Google Scholar]
  19. Kenyon C. J., Walker G. C. DNA-damaging agents stimulate gene expression at specific loci in Escherichia coli. Proc Natl Acad Sci U S A. 1980 May;77(5):2819–2823. doi: 10.1073/pnas.77.5.2819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kondo S. Environmental mutagen testing in Escherichia coli and phage lambda. Mutat Res. 1974 Aug;26(4):235–241. doi: 10.1016/s0027-5107(74)80020-5. [DOI] [PubMed] [Google Scholar]
  21. LWOFF A. Lysogeny. Bacteriol Rev. 1953 Dec;17(4):269–337. doi: 10.1128/br.17.4.269-337.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Levine A., Moreau P. L., Sedgwick S. G., Devoret R. Expression of a bacterial gene turned on by a potent carcinogen. Mutat Res. 1978 Apr;50(1):29–35. doi: 10.1016/0027-5107(78)90057-x. [DOI] [PubMed] [Google Scholar]
  23. Little J. W., Edmiston S. H., Pacelli L. Z., Mount D. W. Cleavage of the Escherichia coli lexA protein by the recA protease. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3225–3229. doi: 10.1073/pnas.77.6.3225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McCann J., Choi E., Yamasaki E., Ames B. N. Detection of carcinogens as mutagens in the Salmonella/microsome test: assay of 300 chemicals. Proc Natl Acad Sci U S A. 1975 Dec;72(12):5135–5139. doi: 10.1073/pnas.72.12.5135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Miller H. I., Kirk M., Echols H. SOS induction and autoregulation of the himA gene for site-specific recombination in Escherichia coli. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6754–6758. doi: 10.1073/pnas.78.11.6754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Moreau P., Bailone A., Devoret R. Prophage lambda induction of Escherichia coli K12 envA uvrB: a highly sensitive test for potential carcinogens. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3700–3704. doi: 10.1073/pnas.73.10.3700. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Radman M. Is there SOS induction in mammalian cells? Photochem Photobiol. 1980 Dec;32(6):823–830. doi: 10.1111/j.1751-1097.1980.tb04062.x. [DOI] [PubMed] [Google Scholar]
  28. Roberts J. W., Roberts C. W., Craig N. L. Escherichia coli recA gene product inactivates phage lambda repressor. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4714–4718. doi: 10.1073/pnas.75.10.4714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shanabruch W. G., Walker G. C. Localization of the plasmid (pKM101) gene(s) involved in recA+lexA+-dependent mutagenesis. Mol Gen Genet. 1980;179(2):289–297. doi: 10.1007/BF00425456. [DOI] [PubMed] [Google Scholar]
  30. TORRIANI A., ROTHMAN F. Mutants of Escherichia coli constitutive for alkaline phosphatase. J Bacteriol. 1961 May;81:835–836. doi: 10.1128/jb.81.5.835-836.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weill-Thevenet N., Buisson J. P., Royer R., Hofnung M. Genetic toxicology studies with 2-nitrobenzofurans and 2-nitronaphthofurans. Mutat Res. 1982 Apr;104(1-3):1–8. doi: 10.1016/0165-7992(82)90112-9. [DOI] [PubMed] [Google Scholar]
  32. Witkin E. M. Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol Rev. 1976 Dec;40(4):869–907. doi: 10.1128/br.40.4.869-907.1976. [DOI] [PMC free article] [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