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. 1980 Apr;142(1):153–161. doi: 10.1128/jb.142.1.153-161.1980

Inhibition of deoxyribonucleic acid gyrase: effects on nucleic acid synthesis and cell division in Escherichia coli K-12.

N F Fairweather, E Orr, I B Holland
PMCID: PMC293920  PMID: 6154685

Abstract

Mutants of Escherichia coli resistant to the antibiotic clorobiocin are also coumermycin resistant, and the mutation to resistance in at least one mutant was mapped near gyrB. We conclude, therefore, that clorobiocin inhibits deoxyribonucleic acid gyrase, and the drug was used to probe the role of this enzyme in vivo. Deozyribonucleic acid synthesis was preferentially inhibited but not completely blocked by the antibiotic. Transcription and cell division were also markedly affected. However, unlike other inhibitors of deoxyribonucleic acid synthesis, clorobiocin failed to induce the synthesis of protein X, the recA gene product. In mutants resistant to clorobiocin the replication velocity was unaffected, but initiation of deoxyribonucleic acid synthesis appeared to be delayed. We conclude that deoxyribonucleic acid gyrase, and hence the supercoiled structure of the chromosome, is important for transcription, normal initiation of deoxyribonucleic acid replication, and cell division. The possible role of deoxyribonucleic acid gyrase in the elongation of replication forks is also discussed.

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Selected References

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  1. Alberts B., Sternglanz R. Recent excitement in the DNA replication problem. Nature. 1977 Oct 20;269(5630):655–661. doi: 10.1038/269655a0. [DOI] [PubMed] [Google Scholar]
  2. Bachmann B. J., Low K. B., Taylor A. L. Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976 Mar;40(1):116–167. doi: 10.1128/br.40.1.116-167.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Botchan P., Wang J. C., Echols H. Effect of circularity and superhelicity on transcription from bacteriophagelambda DNA. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3077–3081. doi: 10.1073/pnas.70.11.3077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boyd A., Holland I. B. Regulation of the synthesis of surface protein in the cell cycle of E. coli B/r. Cell. 1979 Oct;18(2):287–296. doi: 10.1016/0092-8674(79)90048-5. [DOI] [PubMed] [Google Scholar]
  5. Churchward G. G., Holland I. B. Envelope synthesis during the cell cycle in Escherichia coli B/r. J Mol Biol. 1976 Aug 5;105(2):245–261. doi: 10.1016/0022-2836(76)90110-8. [DOI] [PubMed] [Google Scholar]
  6. Cooper S., Helmstetter C. E. Chromosome replication and the division cycle of Escherichia coli B/r. J Mol Biol. 1968 Feb 14;31(3):519–540. doi: 10.1016/0022-2836(68)90425-7. [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. Gellert M., Mizuuchi K., O'Dea M. H., Itoh T., Tomizawa J. I. Nalidixic acid resistance: a second genetic character involved in DNA gyrase activity. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4772–4776. doi: 10.1073/pnas.74.11.4772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gellert M., Mizuuchi K., O'Dea M. H., Nash H. A. DNA gyrase: an enzyme that introduces superhelical turns into DNA. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3872–3876. doi: 10.1073/pnas.73.11.3872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gellert M., O'Dea M. H., Itoh T., Tomizawa J. Novobiocin and coumermycin inhibit DNA supercoiling catalyzed by DNA gyrase. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4474–4478. doi: 10.1073/pnas.73.12.4474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hansen F. G., von Meyenburg K. Characterization of the dnaA, gyrB and other genes in the dnaA region of the Escherichia coli chromosome on specialized transducing phages lambda tna. Mol Gen Genet. 1979 Sep;175(2):135–144. doi: 10.1007/BF00425529. [DOI] [PubMed] [Google Scholar]
  12. Helmstetter C. E., Pierucci O. Cell division during inhibition of deoxyribonucleic acid synthesis in Escherichia coli. J Bacteriol. 1968 May;95(5):1627–1633. doi: 10.1128/jb.95.5.1627-1633.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Helmstetter C. E., Pierucci O. DNA synthesis during the division cycle of three substrains of Escherichia coli B/r. J Mol Biol. 1976 Apr 15;102(3):477–486. doi: 10.1016/0022-2836(76)90329-6. [DOI] [PubMed] [Google Scholar]
  14. Higgins N. P., Peebles C. L., Sugino A., Cozzarelli N. R. Purification of subunits of Escherichia coli DNA gyrase and reconstitution of enzymatic activity. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1773–1777. doi: 10.1073/pnas.75.4.1773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kreuzer K. N., McEntee K., Geballe A. P., Cozzarelli N. R. Lambda transducing phages for the nalA gene of Escherichia coli and conditional lethal nalA mutations. Mol Gen Genet. 1978 Nov 29;167(2):129–137. doi: 10.1007/BF00266906. [DOI] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Marians K. J., Ikeda J. E., Schlagman S., Hurwitz J. Role of DNA gyrase in phiX replicative-form replication in vitro. Proc Natl Acad Sci U S A. 1977 May;74(5):1965–1968. doi: 10.1073/pnas.74.5.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McEntee K. Protein X is the product of the recA gene of Escherichia coli. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5275–5279. doi: 10.1073/pnas.74.12.5275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mizuuchi K., Gellert M., Nash H. A. Involement of supertwisted DNA in integrative recombination of bacteriophage lambda. J Mol Biol. 1978 May 25;121(3):375–392. doi: 10.1016/0022-2836(78)90370-4. [DOI] [PubMed] [Google Scholar]
  20. Mizuuchi K., O'Dea M. H., Gellert M. DNA gyrase: subunit structure and ATPase activity of the purified enzyme. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5960–5963. doi: 10.1073/pnas.75.12.5960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Orr E., Fairweather N. F., Holland I. B., Pritchard R. H. Isolation and characterisation of a strain carrying a conditional lethal mutation in the cou gene of Escherichia coli K12. Mol Gen Genet. 1979;177(1):103–112. doi: 10.1007/BF00267259. [DOI] [PubMed] [Google Scholar]
  22. Peebles C. L., Higgins N. P., Kreuzer K. N., Morrison A., Brown P. O., Sugino A., Cozzarelli N. R. Structure and activities of Escherichia coli DNA gyrase. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):41–52. doi: 10.1101/sqb.1979.043.01.008. [DOI] [PubMed] [Google Scholar]
  23. Pritchard R. H. Review lecture on the growth and form of a bacterial cell. Philos Trans R Soc Lond B Biol Sci. 1974 Feb 21;267(886):303–336. doi: 10.1098/rstb.1974.0003. [DOI] [PubMed] [Google Scholar]
  24. Pritchard R. H., Zaritsky A. Effect of thymine concentration on the replication velocity of DNA in a thymineless mutant of Escherichia coli. Nature. 1970 Apr 11;226(5241):126–131. doi: 10.1038/226126a0. [DOI] [PubMed] [Google Scholar]
  25. Ryan M. J. Coumermycin A1: A preferential inhibitor of replicative DNA synthesis in Escherichia coli. I. In vivo characterization. Biochemistry. 1976 Aug 24;15(17):3769–3777. doi: 10.1021/bi00662a020. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. STACEY K. A., SIMSON E. IMPROVED METHOD FOR THE ISOLATION OF THYMINE-REQUIRING MUTANTS OF ESCHERICHIA COLI. J Bacteriol. 1965 Aug;90:554–555. doi: 10.1128/jb.90.2.554-555.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sanzey B. Modulation of gene expression by drugs affecting deoxyribonucleic acid gyrase. J Bacteriol. 1979 Apr;138(1):40–47. doi: 10.1128/jb.138.1.40-47.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Smith C. L., Oishi M. Early events and mechanisms in the induction of bacterial SOS functions: analysis of the phage repressor inactivation process in vivo. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1657–1661. doi: 10.1073/pnas.75.4.1657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Smith D. H., Davis B. D. Mode of action of novobiocin in Escherichia coli. J Bacteriol. 1967 Jan;93(1):71–79. doi: 10.1128/jb.93.1.71-79.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Staudenbauer W. L. Replication of small plasmids in extracts of Escherichia coli. Mol Gen Genet. 1976 Jun 15;145(3):273–280. doi: 10.1007/BF00325823. [DOI] [PubMed] [Google Scholar]
  33. Sugino A., Higgins N. P., Brown P. O., Peebles C. L., Cozzarelli N. R. Energy coupling in DNA gyrase and the mechanism of action of novobiocin. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4838–4842. doi: 10.1073/pnas.75.10.4838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sugino A., Peebles C. L., Kreuzer K. N., Cozzarelli N. R. Mechanism of action of nalidixic acid: purification of Escherichia coli nalA gene product and its relationship to DNA gyrase and a novel nicking-closing enzyme. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4767–4771. doi: 10.1073/pnas.74.11.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sumida-Yasumoto C., Yudelevich A., Hurwitz J. DNA synthesis in vitro dependent upon phiX174 replicative form I DNA. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1887–1891. doi: 10.1073/pnas.73.6.1887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. 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]

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