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. 1994 May;176(10):3072–3075. doi: 10.1128/jb.176.10.3072-3075.1994

gyrB mutations in coumermycin A1-resistant Borrelia burgdorferi.

D S Samuels 1, R T Marconi 1, W M Huang 1, C F Garon 1
PMCID: PMC205466  PMID: 8188609

Abstract

We have isolated and characterized mutants of Borrelia burgdorferi that are resistant to the antibiotic coumermycin A1, which targets the B subunit of DNA gyrase. Mutants had either 100- or 300-fold higher resistance to coumermycin A1 than wild-type B. burgdorferi. In each case, a single point mutation in the gyrB gene converted Arg-133 to Gly or Ile. Mutations in the homologous Arg residue of Escherichia coli DNA gyrase are also associated with resistance to coumarin antimicrobial agents.

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Selected References

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  1. Ali J. A., Jackson A. P., Howells A. J., Maxwell A. The 43-kilodalton N-terminal fragment of the DNA gyrase B protein hydrolyzes ATP and binds coumarin drugs. Biochemistry. 1993 Mar 16;32(10):2717–2724. doi: 10.1021/bi00061a033. [DOI] [PubMed] [Google Scholar]
  2. Barbour A. G., Garon C. F. Linear plasmids of the bacterium Borrelia burgdorferi have covalently closed ends. Science. 1987 Jul 24;237(4813):409–411. doi: 10.1126/science.3603026. [DOI] [PubMed] [Google Scholar]
  3. Baril C., Richaud C., Baranton G., Saint Girons I. S. Linear chromosome of Borrelia burgdorferi. Res Microbiol. 1989 Oct;140(8):507–516. doi: 10.1016/0923-2508(89)90083-1. [DOI] [PubMed] [Google Scholar]
  4. Burgdorfer W., Barbour A. G., Hayes S. F., Benach J. L., Grunwaldt E., Davis J. P. Lyme disease-a tick-borne spirochetosis? Science. 1982 Jun 18;216(4552):1317–1319. doi: 10.1126/science.7043737. [DOI] [PubMed] [Google Scholar]
  5. Casjens S., Huang W. M. Linear chromosomal physical and genetic map of Borrelia burgdorferi, the Lyme disease agent. Mol Microbiol. 1993 May;8(5):967–980. doi: 10.1111/j.1365-2958.1993.tb01641.x. [DOI] [PubMed] [Google Scholar]
  6. Cinco M. Selection of a Borrelia burgdorferi antigenic variant by cultivation in the presence of increasing amounts of homologous immune serum. FEMS Microbiol Lett. 1992 Apr 1;71(1):15–18. doi: 10.1016/0378-1097(92)90534-u. [DOI] [PubMed] [Google Scholar]
  7. Coleman J. L., Rogers R. C., Benach J. L. Selection of an escape variant of Borrelia burgdorferi by use of bactericidal monoclonal antibodies to OspB. Infect Immun. 1992 Aug;60(8):3098–3104. doi: 10.1128/iai.60.8.3098-3104.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Contreras A., Maxwell A. gyrB mutations which confer coumarin resistance also affect DNA supercoiling and ATP hydrolysis by Escherichia coli DNA gyrase. Mol Microbiol. 1992 Jun;6(12):1617–1624. doi: 10.1111/j.1365-2958.1992.tb00886.x. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Davidson B. E., MacDougall J., Saint Girons I. Physical map of the linear chromosome of the bacterium Borrelia burgdorferi 212, a causative agent of Lyme disease, and localization of rRNA genes. J Bacteriol. 1992 Jun;174(11):3766–3774. doi: 10.1128/jb.174.11.3766-3774.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Drlica K. Control of bacterial DNA supercoiling. Mol Microbiol. 1992 Feb;6(4):425–433. doi: 10.1111/j.1365-2958.1992.tb01486.x. [DOI] [PubMed] [Google Scholar]
  12. Drlica K., Franco R. J. Inhibitors of DNA topoisomerases. Biochemistry. 1988 Apr 5;27(7):2253–2259. doi: 10.1021/bi00407a001. [DOI] [PubMed] [Google Scholar]
  13. Fairweather N. F., Orr E., Holland I. B. Inhibition of deoxyribonucleic acid gyrase: effects on nucleic acid synthesis and cell division in Escherichia coli K-12. J Bacteriol. 1980 Apr;142(1):153–161. doi: 10.1128/jb.142.1.153-161.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ferdows M. S., Barbour A. G. Megabase-sized linear DNA in the bacterium Borrelia burgdorferi, the Lyme disease agent. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5969–5973. doi: 10.1073/pnas.86.15.5969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gellert M. DNA topoisomerases. Annu Rev Biochem. 1981;50:879–910. doi: 10.1146/annurev.bi.50.070181.004311. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Hinnebusch J., Tilly K. Linear plasmids and chromosomes in bacteria. Mol Microbiol. 1993 Dec;10(5):917–922. doi: 10.1111/j.1365-2958.1993.tb00963.x. [DOI] [PubMed] [Google Scholar]
  18. Holmes M. L., Dyall-Smith M. L. Mutations in DNA gyrase result in novobiocin resistance in halophilic archaebacteria. J Bacteriol. 1991 Jan;173(2):642–648. doi: 10.1128/jb.173.2.642-648.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hyde F. W., Johnson R. C. Characterization of a circular plasmid from Borrelia burgdorferi, etiologic agent of Lyme disease. J Clin Microbiol. 1988 Oct;26(10):2203–2205. doi: 10.1128/jcm.26.10.2203-2205.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kurtti T. J., Munderloh U. G., Johnson R. C., Ahlstrand G. G. Colony formation and morphology in Borrelia burgdorferi. J Clin Microbiol. 1987 Nov;25(11):2054–2058. doi: 10.1128/jcm.25.11.2054-2058.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Margolis N., Rosa P. A. Regulation of expression of major outer surface proteins in Borrelia burgdorferi. Infect Immun. 1993 May;61(5):2207–2210. doi: 10.1128/iai.61.5.2207-2210.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maxwell A. The interaction between coumarin drugs and DNA gyrase. Mol Microbiol. 1993 Aug;9(4):681–686. doi: 10.1111/j.1365-2958.1993.tb01728.x. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Old I. G., Margarita D., Saint Girons I. Unique genetic arrangement in the dnaA region of the Borrelia burgdorferi linear chromosome: nucleotide sequence of the dnaA gene. FEMS Microbiol Lett. 1993 Jul 15;111(1):109–114. doi: 10.1111/j.1574-6968.1993.tb06369.x. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Reece R. J., Maxwell A. DNA gyrase: structure and function. Crit Rev Biochem Mol Biol. 1991;26(3-4):335–375. doi: 10.3109/10409239109114072. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Sadziene A., Thomas D. D., Bundoc V. G., Holt S. C., Barbour A. G. A flagella-less mutant of Borrelia burgdorferi. Structural, molecular, and in vitro functional characterization. J Clin Invest. 1991 Jul;88(1):82–92. doi: 10.1172/JCI115308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Samuels D. S., Garon C. F. Coumermycin A1 inhibits growth and induces relaxation of supercoiled plasmids in Borrelia burgdorferi, the Lyme disease agent. Antimicrob Agents Chemother. 1993 Jan;37(1):46–50. doi: 10.1128/aac.37.1.46. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Samuels D. S., Marconi R. T., Garon C. F. Variation in the size of the ospA-containing linear plasmid, but not the linear chromosome, among the three Borrelia species associated with Lyme disease. J Gen Microbiol. 1993 Oct;139(10):2445–2449. doi: 10.1099/00221287-139-10-2445. [DOI] [PubMed] [Google Scholar]
  31. Staudenbauer W. L., Orr E. DNA gyrase: affinity chromatography on novobiocin-Sepharose and catalytic properties. Nucleic Acids Res. 1981 Aug 11;9(15):3589–3603. doi: 10.1093/nar/9.15.3589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Steere A. C. Lyme disease. N Engl J Med. 1989 Aug 31;321(9):586–596. doi: 10.1056/NEJM198908313210906. [DOI] [PubMed] [Google Scholar]
  33. Stålhammar-Carlemalm M., Jenny E., Gern L., Aeschlimann A., Meyer J. Plasmid analysis and restriction fragment length polymorphisms of chromosomal DNA allow a distinction between Borrelia burgdorferi strains. Zentralbl Bakteriol. 1990 Oct;274(1):28–39. doi: 10.1016/s0934-8840(11)80972-2. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Sădziene A., Rosa P. A., Thompson P. A., Hogan D. M., Barbour A. G. Antibody-resistant mutants of Borrelia burgdorferi: in vitro selection and characterization. J Exp Med. 1992 Sep 1;176(3):799–809. doi: 10.1084/jem.176.3.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Thiara A. S., Cundliffe E. Expression and analysis of two gyrB genes from the novobiocin producer, Streptomyces sphaeroides. Mol Microbiol. 1993 May;8(3):495–506. doi: 10.1111/j.1365-2958.1993.tb01593.x. [DOI] [PubMed] [Google Scholar]
  37. Wigley D. B., Davies G. J., Dodson E. J., Maxwell A., Dodson G. Crystal structure of an N-terminal fragment of the DNA gyrase B protein. Nature. 1991 Jun 20;351(6328):624–629. doi: 10.1038/351624a0. [DOI] [PubMed] [Google Scholar]
  38. Zimmer C., Störl K., Störl J. Microbial DNA topoisomerases and their inhibition by antibiotics. J Basic Microbiol. 1990;30(3):209–224. doi: 10.1002/jobm.3620300312. [DOI] [PubMed] [Google Scholar]
  39. del Castillo I., Vizán J. L., Rodríguez-Sáinz M. C., Moreno F. An unusual mechanism for resistance to the antibiotic coumermycin A1. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8860–8864. doi: 10.1073/pnas.88.19.8860. [DOI] [PMC free article] [PubMed] [Google Scholar]

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