Quinolone resistance-determining region in the DNA gyrase gyrA gene of Escherichia coli

Yoshida, Hiroaki; Bogaki, M; Nakamura, Mami; Nakamura, Shinichi

doi:10.1128/aac.34.6.1271

Cited by 606 publications

(568 citation statements)

References 15 publications

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“…The E. coli strains used in the study were DM4100 (30) and the following derivatives of DM4100 constructed by P1-mediated transduction: KD1373 (ParC Ser-80 to Leu), KD2750 (GyrA Ser-83 to Leu and Asp-87 to Tyr), and KD2329 (GyrA Ser-83 to Leu, Asp-87 to Tyr, and ParC Ser-80 to Leu). Introduction of the mutations was confirmed by nucleotide sequence determination of the quinolone resistance-determining regions for gyrA and parC (10,34). Strains were grown on LB agar (23) or in LB liquid medium containing 1% glucose and 100 mM equimolar sodium phosphate buffer (pH 6.8).…”

Section: Methodsmentioning

confidence: 99%

Effect of Anaerobic Growth on Quinolone Lethality with Escherichia coli

Malik¹,

Hussain²,

Drlica³

2007

Antimicrob Agents Chemother

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Quinolone activity against Escherichia coli was examined during aerobic growth, aerobic treatment with chloramphenicol, and anaerobic growth. Nalidixic acid, norfloxacin, ciprofloxacin, and PD161144 were lethal for cultures growing aerobically, and the bacteriostatic activity of each quinolone was unaffected by anaerobic growth. However, lethal activity was distinct for each quinolone with cells treated aerobically with chloramphenicol or grown anaerobically. Nalidixic acid failed to kill cells under both conditions; norfloxacin killed cells when they were grown anaerobically but not when they were treated with chloramphenicol; ciprofloxacin killed cells under both conditions but required higher concentrations than those required with cells grown aerobically; and PD161144, a C-8-methoxy fluoroquinolone, was equally lethal under all conditions. Following pretreatment with nalidixic acid, a shift to anaerobic conditions or the addition of chloramphenicol rapidly blocked further cell death. Formation of quinolone-gyrase-DNA complexes, observed as a sodium dodecyl sulfate (SDS)-dependent drop in cell lysate viscosity, occurred during aerobic and anaerobic growth and in the presence and in the absence of chloramphenicol. However, lethal chromosome fragmentation, detected as a drop in viscosity in the absence of SDS, occurred with nalidixic acid treatment only under aerobic conditions in the absence of chloramphenicol. With PD161144, chromosome fragmentation was detected when the cells were grown aerobically and anaerobically and in the presence and in the absence of chloramphenicol. Thus, all quinolones tested appear to form reversible bacteriostatic complexes containing broken DNA during aerobic growth, during anaerobic growth, and when protein synthesis is blocked; however, the ability to fragment chromosomes and to rapidly kill cells under these conditions depends on quinolone structure.When DNA topoisomerases bind to bacterial DNA, they form complexes that are trapped by the quinolone antibacterials (for a review, see reference 6). These ternary drug-enzyme-DNA complexes block DNA replication (5, 27, 29, 32), RNA synthesis (22, 33), and cell growth (2, 12). A key feature of the complexes is that the trapped DNA moiety is broken, with its ends constrained through covalent linkage to the GyrA protein (24, 25). As a result, quinolone concentrations sufficient to block replication do not relax chromosomal DNA supercoiling (29). When the quinolone is removed, the breaks are readily resealed (11, 31) and the inhibitory effects of the compounds are reversed (13,21,27). Irreversible events that lead to rapid cell death occur at higher quinolone concentrations (1, 20). We have proposed that rapid cell death arises from chromosome fragmentation that occurs when doublestrand DNA breaks are released from the protein-mediated constraint present in ternary complexes. At rapidly lethal quinolone concentrations, supercoils cannot be maintained (1) and fragmented DNA is obtained under conditions that allow resealing at bacteriost...

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Section: Methodsmentioning

confidence: 99%

Effect of Anaerobic Growth on Quinolone Lethality with Escherichia coli

Malik¹,

Hussain²,

Drlica³

2007

Antimicrob Agents Chemother

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“…Recently, the two genes that code for the subunits of topoisomerase IV,parC and parE, have been cloned (16), and strains which overexpress both subunits have been constructed (18,27 (41), implies that quinolones may be capable of inhibiting the activity of topoisomerase IV as well as against DNA gyrase. In this study, in order to define the inhibitory activities of quinolones against topoisomerase IV, both subunits of topoisomerase IV, ParC and ParE, were purified, and optimum conditions for the decatenating activity of topoisomerase IV were determined.…”

mentioning

confidence: 99%

Comparison of inhibition of Escherichia coli topoisomerase IV by quinolones with DNA gyrase inhibition

Hoshino¹,

Kitamura²,

Morrissey³

et al. 1994

Antimicrob Agents Chemother

153

100

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In order to examine the inhibitory activities of quinolones against topoisomerase IV, both subunits of this enzyme, ParC and ParE, were purified from Escherichia coli. The specific activity of topoisomerase IV decatenation was found to be more than five times greater than that of topoisomerase IV relaxation. Thus, the decatenation activity of topoisomerase IV seems the most relevant activity for use in studies of drug inhibition of this enzyme. Although topoisomerase IV was less sensitive to quinolones than DNA gyrase, the 50% inhibitory concentrations for decatenation were significantly lower than those for type I topoisomerases. Moreover, there was a positive correlation between the inhibitory activity against topoisomerase IV decatenation and that for DNA gyrase supercoiling. These results imply that topoisomerase IV could be a target for the quinolones in intact bacteria and that quinolones could inhibit not only supercoiling of DNA gyrase but also decatenation of topoisomerase IV when high concentrations of drug exist in bacterial cells.Four topoisomerases have been isolated from Escherichia coli so far: topoisomerase I (39), topoisomerase II (DNA gyrase) (9), topoisomerase III (34), and topoisomerase IV (16). Among these enzymes, topoisomerases I, II, and IV are implicated in the control of the intracellular supercoiling density of chromosomal or plasmid DNA, affecting the efficacy of DNA replication and transcription (4, 6-8, 20, 24, 29, 36, 37). In addition, some topoisomerases are required for the segregation of daughter chromosomes or plasmid DNA (1,21,35,40). Although all of these topoisomerases are capable of changing the topology of DNA by a cleaving and rejoining step, each enzyme appears to possess a favored reaction in vitro. Type I topoisomerases, topoisomerase I and topoisomerase III, show efficient relaxing and decatenating activity, respectively, in the absence of ATP (2,3,5,11,29,30,34 subunits of DNA gyrase), respectively (16). Similar sequences are located especially around the region of DNA gyrase known as the "quinolone resistance-determining region." The quinolone antibacterial agents, in particular the fluoroquinolones, strongly inhibit DNA gyrase, which results ultimately in bacterial death (32). However, the inhibitory activities of quinolones against type I topoisomerases, topoisomerase I (25) and topoisomerase III (2) in E. coli, are weak. The inhibitory activities of fluoroquinolones against topoisomerase IV have yet to be fully investigated. The similarity in amino acid sequence between DNA gyrase and topoisomerase IV, especially around sites in the GyrA protein of DNA gyrase at positions known to produce quinolone-resistant DNA gyrases (41), implies that quinolones may be capable of inhibiting the activity of topoisomerase IV as well as against DNA gyrase.In this study, in order to define the inhibitory activities of quinolones against topoisomerase IV, both subunits of topoisomerase IV, ParC and ParE, were purified, and optimum conditions for the decatenating activity of topoi...

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“…16,17 In conclusion, ampicillin, chloramphenicol and cotrimoxazole are no longer used in empiric treatment.The alternatives for acute diarrheal infection is third generation cephalosporins and its limited uses in pediatric patients, as it is not recommended below the age of 14 years old; reducing the choices to a very narrow scale during the treatment of acute bacterial gastroenteritis, but still, the prevention is the best treatment of such case indeed.…”

Section: Discussionmentioning

confidence: 99%

The incidence of Shigella and Salmonella in the stool of pediatric patients

2017

Iraqi J. Public Health

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Objective: Gastroenteritis is a common illness in the pediatric age group, the causes could be viral, bacterial and fungal. This study was focused on bacterial pathogens like Shigella and Salmonella.Methods: About 1399 stool samples were collected in a period between January 2010 and May 2011 at the child welfare hospital, all the samples were cultured on suitable culture media and then biochemical activities were tested using API-E 20 and sugar fermentation tests, the isolates were sent for sensitivity test with a group of antibiotics. Results:Out of 1399 cases 50 cases only showed a positive growth for Shigella and Salmonella as pathological microorganisms. Conclusion:The incidence of Shigella and Salmonella was considered low when compared to other causes of gastroenteritis among children as enteropathogenic Escherichia coli or viral causes.

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Quinolone resistance-determining region in the DNA gyrase gyrA gene of Escherichia coli

Cited by 606 publications

References 15 publications

Effect of Anaerobic Growth on Quinolone Lethality with Escherichia coli

Effect of Anaerobic Growth on Quinolone Lethality with Escherichia coli

Comparison of inhibition of Escherichia coli topoisomerase IV by quinolones with DNA gyrase inhibition

The incidence of Shigella and Salmonella in the stool of pediatric patients

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