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Proc Natl Acad Sci U S A. Dec 5, 1995; 92(25): 11801–11805.

Topoisomerase IV is a target of quinolones in Escherichia coli.


We have demonstrated that, in Escherichia coli, quinolone antimicrobial agents target topoisomerase IV (topo IV). The inhibition of topo IV becomes apparent only when gyrase is mutated to quinolone resistance. In such mutants, these antibiotics caused accumulation of replication catenanes, which is diagnostic of a loss of topo IV activity. Mutant forms of topo IV provided an additional 10-fold resistance to quinolones and prevented drug-induced catenane accumulation. Drug inhibition of topo IV differs from that of gyrase. (i) Wild-type topo IV is not dominant over the resistant allele. (ii) Inhibition of topo IV leads to only a slow stop in replication. (iii) Inhibition of topo IV is primarily bacteriostatic. These differences may result from topo IV acting behind the replication fork, allowing for repair of drug-induced lesions. We suggest that this and a slightly higher intrinsic resistance of topo IV make it secondary to gyrase as a quinolone target. Our results imply that the quinolone binding pockets of gyrase and topo IV are similar and that substantial levels of drug resistance require mutations in both enzymes.

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Selected References

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  • Sugino A, Peebles CL, Kreuzer KN, Cozzarelli NR. 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. [PMC free article] [PubMed]
  • Gellert M, Mizuuchi K, O'Dea MH, Itoh T, Tomizawa JI. 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. [PMC free article] [PubMed]
  • Wang JC. DNA topoisomerases: why so many? J Biol Chem. 1991 Apr 15;266(11):6659–6662. [PubMed]
  • Kreuzer KN, Cozzarelli NR. 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. [PMC free article] [PubMed]
  • Tewey KM, Rowe TC, Yang L, Halligan BD, Liu LF. Adriamycin-induced DNA damage mediated by mammalian DNA topoisomerase II. Science. 1984 Oct 26;226(4673):466–468. [PubMed]
  • Howard MT, Neece SH, Matson SW, Kreuzer KN. Disruption of a topoisomerase-DNA cleavage complex by a DNA helicase. Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):12031–12035. [PMC free article] [PubMed]
  • Heisig P, Schedletzky H, Falkenstein-Paul H. Mutations in the gyrA gene of a highly fluoroquinolone-resistant clinical isolate of Escherichia coli. Antimicrob Agents Chemother. 1993 Apr;37(4):696–701. [PMC free article] [PubMed]
  • 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. [PubMed]
  • Adams DE, Shekhtman EM, Zechiedrich EL, Schmid MB, Cozzarelli NR. 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. [PubMed]
  • Kato J, Suzuki H, Ikeda H. Purification and characterization of DNA topoisomerase IV in Escherichia coli. J Biol Chem. 1992 Dec 25;267(36):25676–25684. [PubMed]
  • 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. [PubMed]
  • Belland RJ, Morrison SG, Ison C, Huang WM. Neisseria gonorrhoeae acquires mutations in analogous regions of gyrA and parC in fluoroquinolone-resistant isolates. Mol Microbiol. 1994 Oct;14(2):371–380. [PubMed]
  • Heisig P, Wiedemann B. Use of a broad-host-range gyrA plasmid for genetic characterization of fluoroquinolone-resistant gram-negative bacteria. Antimicrob Agents Chemother. 1991 Oct;35(10):2031–2036. [PMC free article] [PubMed]
  • Winans SC, Elledge SJ, Krueger JH, Walker GC. Site-directed insertion and deletion mutagenesis with cloned fragments in Escherichia coli. J Bacteriol. 1985 Mar;161(3):1219–1221. [PMC free article] [PubMed]
  • Mizuuchi K, Mizuuchi M, O'Dea MH, Gellert M. Cloning and simplified purification of Escherichia coli DNA gyrase A and B proteins. J Biol Chem. 1984 Jul 25;259(14):9199–9201. [PubMed]
  • Peng H, Marians KJ. Escherichia coli topoisomerase IV. Purification, characterization, subunit structure, and subunit interactions. J Biol Chem. 1993 Nov 15;268(32):24481–24490. [PubMed]
  • Otter R, Cozzarelli NR. Escherichia coli DNA gyrase. Methods Enzymol. 1983;100:171–180. [PubMed]
  • Shin CG, Strayer JM, Wani MA, Snapka RM. Rapid evaluation of topoisomerase inhibitors: caffeine inhibition of topoisomerases in vivo. Teratog Carcinog Mutagen. 1990;10(1):41–52. [PubMed]
  • Shishido K, Komiyama N, Ikawa S. Increased production of a knotted form of plasmid pBR322 DNA in Escherichia coli DNA topoisomerase mutants. J Mol Biol. 1987 May 5;195(1):215–218. [PubMed]
  • Hane MW, Wood TH. Escherichia coli K-12 mutants resistant to nalidixic acid: genetic mapping and dominance studies. J Bacteriol. 1969 Jul;99(1):238–241. [PMC free article] [PubMed]
  • Liu YX, Hsiung Y, Jannatipour M, Yeh Y, Nitiss JL. Yeast topoisomerase II mutants resistant to anti-topoisomerase agents: identification and characterization of new yeast topoisomerase II mutants selected for resistance to etoposide. Cancer Res. 1994 Jun 1;54(11):2943–2951. [PubMed]
  • Tanaka M, Sato K, Kimura Y, Hayakawa I, Osada Y, Nishino T. Inhibition by quinolones of DNA gyrase from Staphylococcus aureus. Antimicrob Agents Chemother. 1991 Jul;35(7):1489–1491. [PMC free article] [PubMed]

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