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. 1996 Jul;40(7):1594–1599. doi: 10.1128/aac.40.7.1594

Fluoroquinolone action in mycobacteria: similarity with effects in Escherichia coli and detection by cell lysate viscosity.

K Drlica 1, C Xu 1, J Y Wang 1, R M Burger 1, M Malik 1
PMCID: PMC163379  PMID: 8807046

Abstract

Fluoroquinolones are potent antibacterial agents that are being used as therapeutic agents for the treatment of multidrug-resistant tuberculosis. To better understand fluoroquinolone action in mycobacteria, the effects of ciprofloxacin were examined. DNA synthesis was inhibited rapidly in Mycobacterium smegmatis, DNA cleavage was readily observed by an empirical assay of cell lysate viscosity, and cell growth was blocked. These data are explained by the formation of gyrase-DNA-ciprofloxacin complexes that block replication fork movement. The bactericidal action of ciprofloxacin against M. smegmatis, Mycobacterium bovis BCG, and Escherichia coli occurred more slowly in cells with longer doubling times. The bactericidal effect against M. bovis BCG was partially blocked by pretreatment with chloramphenicol, an inhibitor of protein synthesis, and by very high concentrations of ciprofloxacin itself. Similar responses occur when E. coli is treated with ciprofloxacin. These similarities between E. coli and mycobacteria indicate that results from extensive fluoroquinolone studies with E. coli can be applied to mycobacteria. A simple viscometric assay of DNA cleavage is described. The assay is expected to be useful for screening new fluoroquinolone derivatives for increased effectiveness against clinically important bacteria.

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Selected References

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  1. Adams D. E., Shekhtman E. M., Zechiedrich E. L., Schmid M. B., Cozzarelli N. R. The role of topoisomerase IV in partitioning bacterial replicons and the structure of catenated intermediates in DNA replication. Cell. 1992 Oct 16;71(2):277–288. doi: 10.1016/0092-8674(92)90356-h. [DOI] [PubMed] [Google Scholar]
  2. Chow R. T., Dougherty T. J., Fraimow H. S., Bellin E. Y., Miller M. H. Association between early inhibition of DNA synthesis and the MICs and MBCs of carboxyquinolone antimicrobial agents for wild-type and mutant [gyrA nfxB(ompF) acrA] Escherichia coli K-12. Antimicrob Agents Chemother. 1988 Aug;32(8):1113–1118. doi: 10.1128/aac.32.8.1113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Crumplin G. C., Smith J. T. Nalidixic acid: an antibacterial paradox. Antimicrob Agents Chemother. 1975 Sep;8(3):251–261. doi: 10.1128/aac.8.3.251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Deitz W. H., Cook T. M., Goss W. A. Mechanism of action of nalidixic acid on Escherichia coli. 3. Conditions required for lethality. J Bacteriol. 1966 Feb;91(2):768–773. doi: 10.1128/jb.91.2.768-773.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Drlica K., Engle E. C., Manes S. H. DNA gyrase on the bacterial chromosome: possibility of two levels of action. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6879–6883. doi: 10.1073/pnas.77.11.6879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Drlica K., Worcel A. Conformational transitions in the Escherichia coli chromosome: analysis by viscometry and sedimentation. J Mol Biol. 1975 Oct 25;98(2):393–411. doi: 10.1016/s0022-2836(75)80126-4. [DOI] [PubMed] [Google Scholar]
  8. Engle E. C., Manes S. H., Drlica K. Differential effects of antibiotics inhibiting gyrase. J Bacteriol. 1982 Jan;149(1):92–98. doi: 10.1128/jb.149.1.92-98.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ferrero L., Cameron B., Crouzet J. Analysis of gyrA and grlA mutations in stepwise-selected ciprofloxacin-resistant mutants of Staphylococcus aureus. Antimicrob Agents Chemother. 1995 Jul;39(7):1554–1558. doi: 10.1128/aac.39.7.1554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ferrero L., Cameron B., Manse B., Lagneaux D., Crouzet J., Famechon A., Blanche F. Cloning and primary structure of Staphylococcus aureus DNA topoisomerase IV: a primary target of fluoroquinolones. Mol Microbiol. 1994 Aug;13(4):641–653. doi: 10.1111/j.1365-2958.1994.tb00458.x. [DOI] [PubMed] [Google Scholar]
  11. Frieden T. R., Sterling T., Pablos-Mendez A., Kilburn J. O., Cauthen G. M., Dooley S. W. The emergence of drug-resistant tuberculosis in New York City. N Engl J Med. 1993 Feb 25;328(8):521–526. doi: 10.1056/NEJM199302253280801. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Jacobs W. R., Jr, Kalpana G. V., Cirillo J. D., Pascopella L., Snapper S. B., Udani R. A., Jones W., Barletta R. G., Bloom B. R. Genetic systems for mycobacteria. Methods Enzymol. 1991;204:537–555. doi: 10.1016/0076-6879(91)04027-l. [DOI] [PubMed] [Google Scholar]
  15. Kato J., Nishimura Y., Imamura R., Niki H., Hiraga S., Suzuki H. New topoisomerase essential for chromosome segregation in E. coli. Cell. 1990 Oct 19;63(2):393–404. doi: 10.1016/0092-8674(90)90172-b. [DOI] [PubMed] [Google Scholar]
  16. Khodursky A. B., Zechiedrich E. L., Cozzarelli N. R. Topoisomerase IV is a target of quinolones in Escherichia coli. Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11801–11805. doi: 10.1073/pnas.92.25.11801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kreuzer K. N., Cozzarelli N. R. Escherichia coli mutants thermosensitive for deoxyribonucleic acid gyrase subunit A: effects on deoxyribonucleic acid replication, transcription, and bacteriophage growth. J Bacteriol. 1979 Nov;140(2):424–435. doi: 10.1128/jb.140.2.424-435.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lewin C. S., Howard B. M., Smith J. T. Protein- and RNA-synthesis independent bactericidal activity of ciprofloxacin that involves the A subunit of DNA gyrase. J Med Microbiol. 1991 Jan;34(1):19–22. doi: 10.1099/00222615-34-1-19. [DOI] [PubMed] [Google Scholar]
  19. Lewin C. S., Morrissey I., Smith J. T. The mode of action of quinolones: the paradox in activity of low and high concentrations and activity in the anaerobic environment. Eur J Clin Microbiol Infect Dis. 1991 Apr;10(4):240–248. doi: 10.1007/BF01966996. [DOI] [PubMed] [Google Scholar]
  20. Pruss G. J., Franco R. J., Chevalier S. G., Manes S. H., Drlica K. Effects of DNA gyrase inhibitors in Escherichia coli topoisomerase I mutants. J Bacteriol. 1986 Oct;168(1):276–282. doi: 10.1128/jb.168.1.276-282.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Sternglanz R., DiNardo S., Voelkel K. A., Nishimura Y., Hirota Y., Becherer K., Zumstein L., Wang J. C. Mutations in the gene coding for Escherichia coli DNA topoisomerase I affect transcription and transposition. Proc Natl Acad Sci U S A. 1981 May;78(5):2747–2751. doi: 10.1073/pnas.78.5.2747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Sullivan E. A., Kreiswirth B. N., Palumbo L., Kapur V., Musser J. M., Ebrahimzadeh A., Frieden T. R. Emergence of fluoroquinolone-resistant tuberculosis in New York City. Lancet. 1995 May 6;345(8958):1148–1150. doi: 10.1016/s0140-6736(95)90980-x. [DOI] [PubMed] [Google Scholar]
  25. Takiff H. E., Salazar L., Guerrero C., Philipp W., Huang W. M., Kreiswirth B., Cole S. T., Jacobs W. R., Jr, Telenti A. Cloning and nucleotide sequence of Mycobacterium tuberculosis gyrA and gyrB genes and detection of quinolone resistance mutations. Antimicrob Agents Chemother. 1994 Apr;38(4):773–780. doi: 10.1128/aac.38.4.773. [DOI] [PMC free article] [PubMed] [Google Scholar]

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