Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1982 Aug;151(2):764–770. doi: 10.1128/jb.151.2.764-770.1982

Enhanced Resistance to Nitrosoguanidine Killing and Mutagenesis in a DNA Gyrase Mutant of Escherichia coli

Lee Chao 1, D Michael Tillman 1
PMCID: PMC220323  PMID: 6178722

Abstract

The role of DNA gyrase in handling DNA damages induced by N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) was examined with two Escherichia coli strains, KL161 and KL166. The two strains are isogenic except that KL166 harbors a mutation at the nalA (gyrA) locus which specifies one of the two subunits of DNA gyrase. We treated the two strains with several different types of mutagenic agents and found the nalA strain to be highly resistant to MNNG-induced killing and mutagenic effects as compared with the parental strain. The MNNG resistance was specific, since the two strains were about equally sensitive to methyl methane sulfonate, ethyl methane sulfonate, and UV and gamma radiations. We pulse-labeled the two strains with [3H]uridine and 14C-amino acids after MNNG treatment to analyze RNA and protein synthetic rates. The pulse-labeled proteins were also separated on polyacrylamide gels. The results show that pulse-labeled RNA and proteins persisted in the nalA strain but declined rapidly in the parental strain after MNNG treatment. We compared membrane-free nucleoid preparations from the two strains by sucrose density gradient centrifugation and found a difference in nucleoid organization between the two strains. The nucleoid of the nalA strain, unlike that of the parental strain, may have a highly ordered structure, as indicated by its resistance to ethidium bromide-induced relaxation. The ability of the two strains to express an adaptive response to MNNG was determined. We found that the resistance to MNNG killing and mutagenesis by the nalA strain cannot be further increased by adaptive treatment. These results suggest that an alteration in DNA gyrase may have profound effects on E. coli chromosome organization and base methylation by MNNG.

Full text

PDF
764

Images in this article

768Image
on p.768

Selected References

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

  1. Anderson T. J., Burdon R. H. N-methyl-N'-nitro-N-nitrosoguanidine: reactions of possible significance to biological activity with mammalian cells. Cancer Res. 1970 Jun;30(6):1773–1781. [PubMed] [Google Scholar]
  2. Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cerdá-Olmedo E., Hanawalt P. C. Macromolecular action of nitrosoguanidine in Escherichia coli. Biochim Biophys Acta. 1967 Jul 18;142(2):450–464. doi: 10.1016/0005-2787(67)90626-0. [DOI] [PubMed] [Google Scholar]
  4. Chao L. Regulation of RNA polymerase subunit synthesis in Escherichia coli: utilization of DNA-Intercalating drugs as a probe. Arch Biochem Biophys. 1977 Sep;183(1):242–249. doi: 10.1016/0003-9861(77)90437-4. [DOI] [PubMed] [Google Scholar]
  5. Cozzarelli N. R. DNA gyrase and the supercoiling of DNA. Science. 1980 Feb 29;207(4434):953–960. doi: 10.1126/science.6243420. [DOI] [PubMed] [Google Scholar]
  6. Drake J. W., Baltz R. H. The biochemistry of mutagenesis. Annu Rev Biochem. 1976;45:11–37. doi: 10.1146/annurev.bi.45.070176.000303. [DOI] [PubMed] [Google Scholar]
  7. Drlica K., Snyder M. Superhelical Escherichia coli DNA: relaxation by coumermycin. J Mol Biol. 1978 Apr 5;120(2):145–154. doi: 10.1016/0022-2836(78)90061-x. [DOI] [PubMed] [Google Scholar]
  8. Gerchman L. L., Ludlum D. B. The properties of O 6 -methylguanine in templates for RNA polymerase. Biochim Biophys Acta. 1973 May 10;308(2):310–316. doi: 10.1016/0005-2787(73)90160-3. [DOI] [PubMed] [Google Scholar]
  9. Guertin M., Mamet-Bratley M. D. Difference in the action of ethyl and methyl methane sulfonates on DNA template activity for RNA synthesis in vitro. Biochim Biophys Acta. 1975 May 16;390(3):312–318. doi: 10.1016/0005-2787(75)90351-2. [DOI] [PubMed] [Google Scholar]
  10. Hays J. B., Boehmer S. Antagonists of DNA gyrase inhibit repair and recombination of UV-irradiated phage lambda. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4125–4129. doi: 10.1073/pnas.75.9.4125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jeggo P., Defais T. M., Samson L., Schendel P. An adaptive response of E. coli to low levels of alkylating agent: comparison with previously characterised DNA repair pathways. Mol Gen Genet. 1977 Nov 29;157(1):1–9. doi: 10.1007/BF00268680. [DOI] [PubMed] [Google Scholar]
  12. Jeggo P. Isolation and characterization of Escherichia coli K-12 mutants unable to induce the adaptive response to simple alkylating agents. J Bacteriol. 1979 Sep;139(3):783–791. doi: 10.1128/jb.139.3.783-791.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johnsrud L. Contacts between Escherichia coli RNA polymerase and a lac operon promoter. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5314–5318. doi: 10.1073/pnas.75.11.5314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kondo S., Ichikawa H. Evidence that pretreatment of Escherichia coli cells with N-methyl-N'-nitro-N-nitrosoguanidine enhances mutability of subsequently infecting phage lambda. Mol Gen Genet. 1973 Nov 22;126(4):319–324. doi: 10.1007/BF00269441. [DOI] [PubMed] [Google Scholar]
  15. Lawley P. D., Martin C. N. Molecular mechanisms in alkylation mutagenesis. Induced reversion of bacteriophage T4rII AP72 by ethyl methanesulphonate in relation to extent and mode of ethylation of purines in bacteriophage deoxyribonucleic acid. Biochem J. 1975 Jan;145(1):85–91. doi: 10.1042/bj1450085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lawley P. D., Thatcher C. J. Methylation of deoxyribonucleic acid in cultured mammalian cells by N-methyl-N'-nitro-N-nitrosoguanidine. The influence of cellular thiol concentrations on the extent of methylation and the 6-oxygen atom of guanine as a site of methylation. Biochem J. 1970 Feb;116(4):693–707. doi: 10.1042/bj1160693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Liu L. F., Wang J. C. DNA-DNA gyrase complex: the wrapping of the DNA duplex outside the enzyme. Cell. 1978 Nov;15(3):979–984. doi: 10.1016/0092-8674(78)90281-7. [DOI] [PubMed] [Google Scholar]
  18. Pegg A. E. Formation and metabolism of alkylated nucleosides: possible role in carcinogenesis by nitroso compounds and alkylating agents. Adv Cancer Res. 1977;25:195–269. doi: 10.1016/s0065-230x(08)60635-1. [DOI] [PubMed] [Google Scholar]
  19. Radman M. SOS repair hypothesis: phenomenology of an inducible DNA repair which is accompanied by mutagenesis. Basic Life Sci. 1975;5A:355–367. doi: 10.1007/978-1-4684-2895-7_48. [DOI] [PubMed] [Google Scholar]
  20. Robins P., Cairns J. Quantitation of the adaptive response to alkylating agents. Nature. 1979 Jul 5;280(5717):74–76. doi: 10.1038/280074a0. [DOI] [PubMed] [Google Scholar]
  21. Ruiz-Vázquez R., Cerdá-Olmedo E. An Escherichia coli mutant refractory to nitrosoguanidine mutagenesis. Mol Gen Genet. 1980;178(3):625–631. doi: 10.1007/BF00337870. [DOI] [PubMed] [Google Scholar]
  22. Ryan M. J., Wells R. D. Coumerimycin A1: A preferential inhibitor of replicative DNA synthesis in Escherichia coli. II. In vivo characterization. Biochemistry. 1976 Aug 24;15(17):3778–3782. doi: 10.1021/bi00662a021. [DOI] [PubMed] [Google Scholar]
  23. Samson L., Cairns J. A new pathway for DNA repair in Escherichia coli. Nature. 1977 May 19;267(5608):281–283. doi: 10.1038/267281a0. [DOI] [PubMed] [Google Scholar]
  24. Schendel P. F., Robins P. E. Repair of O6-methylguanine in adapted Escherichia coli. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6017–6020. doi: 10.1073/pnas.75.12.6017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Singer B., Fraenkel-Conrat H. Chemical modification of viral ribonucleic acid. VII. The action of methylating agents and nitrosoguanidine on polynucleotides including tobacco mosaic virus ribonucleic acid. Biochemistry. 1969 Aug;8(8):3260–3266. doi: 10.1021/bi00836a019. [DOI] [PubMed] [Google Scholar]
  26. Singer B., Fraenkel-Conrat H., Kuśmierek J. T. Preparation and template activities of polynucleotides containing O2- and O4-alkyluridine. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1722–1726. doi: 10.1073/pnas.75.4.1722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Snyder M., Drlica K. DNA gyrase on the bacterial chromosome: DNA cleavage induced by oxolinic acid. J Mol Biol. 1979 Jun 25;131(2):287–302. doi: 10.1016/0022-2836(79)90077-9. [DOI] [PubMed] [Google Scholar]
  28. Staudenbauer W. L. Letters to the editor: Novobiocin-a specific inhibitor of semiconservative DNA replication in permeabilized Escherichia coli cells. J Mol Biol. 1975 Jul 25;96(1):201–205. doi: 10.1016/0022-2836(75)90191-6. [DOI] [PubMed] [Google Scholar]
  29. Town C. D., Smith K. C., Kaplan H. S. Production and repair of radiochemical damage in Escherichia coli deoxyribonucleic acid; its modification by culture conditions and relation to survival. J Bacteriol. 1971 Jan;105(1):127–135. doi: 10.1128/jb.105.1.127-135.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ulmer K. M., Gomez R. F., Sinskey A. J. Ionizing radiation damage to the folded chromosome of Escherichia coli K-12: sedimentation properties of irradiated nucleoids and chromosomal deoxyribonucleic acid. J Bacteriol. 1979 May;138(2):475–485. doi: 10.1128/jb.138.2.475-485.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Worcel A., Burgi E. On the structure of the folded chromosome of Escherichia coli. J Mol Biol. 1972 Nov 14;71(2):127–147. doi: 10.1016/0022-2836(72)90342-7. [DOI] [PubMed] [Google Scholar]
  33. von Wright A., Bridges B. A. Effect of gyrB-mediated changes in chromosome structure on killing of Escherichia coli by ultraviolet light: experiments with strains differing in deoxyribonucleic acid repair capacity. J Bacteriol. 1981 Apr;146(1):18–23. doi: 10.1128/jb.146.1.18-23.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES