19 DNA Gyrase and Other Type II Topoisomerases

The Enzymes ◽  
1981 ◽  
pp. 345-366 ◽  
Author(s):  
Martin Gellert
Keyword(s):  
Dna Gyrase ◽  
Type Ii

scholarly journals Differential behaviors of Staphylococcus aureus and Escherichia coli type II DNA topoisomerases.

1996 ◽  
Vol 40 (12) ◽  
pp. 2714-2720 ◽  
Author(s):  
F Blanche ◽  
B Cameron ◽  
F X Bernard ◽  
L Maton ◽  
B Manse ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Strong Support ◽  
Dna Gyrase ◽  
Dna Supercoiling ◽  
Type Ii ◽  
Topoisomerase Iv ◽  
E Coli ◽  
Dna Relaxation

Staphylococcus aureus gyrA and gyrB genes encoding DNA gyrase subunits were cloned and coexpressed in Escherichia coli under the control of the T7 promoter-T7 RNA polymerase system, leading to soluble gyrase which was purified to homogeneity. Purified gyrase was catalytically indistinguishable from the gyrase purified from S. aureus and did not contain detectable amounts of topoisomerases from the E. coli host. Topoisomerase IV subunits GrlA and GrlB from S. aureus were also expressed in E. coli and were separately purified to apparent homogeneity. Topoisomerase IV, which was reconstituted by mixing equimolar amounts of GrlA and GrlB, had both ATP-dependent decatenation and DNA relaxation activities in vitro. This enzyme was more sensitive than gyrase to inhibition by typical fluoroquinolone antimicrobial agents such as ciprofloxacin or sparfloxacin, adding strong support to genetic studies which indicate that topoisomerase IV is the primary target of fluoroquinolones in S. aureus. The results obtained with ofloxacin suggest that this fluoroquinolone could also primarily target gyrase. No cleavable complex could be detected with S. aureus gyrase upon incubation with ciprofloxacin or sparfloxacin at concentrations which fully inhibit DNA supercoiling. This suggests that these drugs do not stabilize the open DNA-gyrase complex, at least under standard in vitro incubation conditions, but are more likely to interfere primarily with the DNA breakage step, contrary to what has been reported with E. coli gyrase. Both S. aureus gyrase-catalyzed DNA supercoiling and S. aureus topoisomerase IV-catalyzed decatenation were dramatically stimulated by potassium glutamate or aspartate (500- and 50-fold by 700 and 350 mM glutamate, respectively), whereas topoisomerase IV-dependent DNA relaxation was inhibited 3-fold by 350 mM glutamate. The relevance of the effect of dicarboxylic amino acids on the activities of type II topoisomerases is discussed with regard to the intracellular osmolite composition of S. aureus.

scholarly journals Inhibitory Activities of Gatifloxacin (AM-1155), a Newly Developed Fluoroquinolone, against Bacterial and Mammalian Type II Topoisomerases

1998 ◽  
Vol 42 (10) ◽  
pp. 2678-2681 ◽  
Author(s):  
Masaya Takei ◽  
Hideyuki Fukuda ◽  
Tokutaro Yasue ◽  
Masaki Hosaka ◽  
Yasuo Oomori
Keyword(s):  
Hela Cell ◽  
Topoisomerase Ii ◽  
Dna Gyrase ◽  
Antibacterial Activities ◽  
Type Ii ◽  
Topoisomerase Iv ◽  
E Coli ◽  
Bacterial Enzyme ◽  
Inhibitory Activities ◽  
Bacterial Type

ABSTRACT We determined the inhibitory activities of gatifloxacin againstStaphylococcus aureus topoisomerase IV,Escherichia coli DNA gyrase, and HeLa cell topoisomerase II and compared them with those of several quinolones. The inhibitory activities of quinolones against these type II topoisomerases significantly correlated with their antibacterial activities or cytotoxicities (correlation coefficient [r] = 0.926 forS. aureus, r = 0.972 for E. coli, and r = 0.648 for HeLa cells). Gatifloxacin possessed potent inhibitory activities against bacterial type II topoisomerases (50% inhibitory concentration [IC50] = 13.8 μg/ml for S. aureustopoisomerase IV; IC50 = 0.109 μg/ml for E. coli DNA gyrase) but the lowest activity against HeLa cell topoisomerase II (IC50 = 265 μg/ml) among the quinolones tested. There was also a significant correlation between the inhibitory activities of quinolones against S. aureustopoisomerase IV and those against E. coli DNA gyrase (r = 0.969). However, the inhibitory activity against HeLa cell topoisomerase II did not correlate with that against either bacterial enzyme. The IC50 of gatifloxacin for HeLa cell topoisomerase II was 19 and was more than 2,400 times higher than that for S. aureus topoisomerase IV and that for E. coli DNA gyrase. These ratios were higher than those for other quinolones, indicating that gatifloxacin possesses a higher selectivity for bacterial type II topoisomerases.

In vitroandin vivometabolism of14C-AZ11, a novel inhibitor of bacterial DNA gyrase/type II topoisomerase

Xenobiotica ◽  
2014 ◽  
Vol 45 (2) ◽  
pp. 158-170 ◽  
Author(s):  
Jian Guo ◽  
Camil Joubran ◽  
Ricardo A. Luzietti ◽  
Fei Zhou ◽  
Gregory S. Basarab ◽  
...  
Keyword(s):  
Dna Gyrase ◽  
Type Ii ◽  
Bacterial Dna

scholarly journals Molecular Basis for the Differential Quinolone Susceptibility of Mycobacterial DNA Gyrase

2014 ◽  
Vol 58 (4) ◽  
pp. 2013-2020 ◽  
Author(s):  
Rupesh Kumar ◽  
Bhavani Shankar Madhumathi ◽  
Valakunja Nagaraja
Keyword(s):  
Differential Susceptibility ◽  
Dna Gyrase ◽  
Drug Binding ◽  
Double Strand Breaks ◽  
Type Ii ◽  
Content Type ◽  
Strand Breaks ◽  
E Coli ◽  
Dna Complex ◽  
Negative Supercoils

ABSTRACTDNA gyrase is a type II topoisomerase that catalyzes the introduction of negative supercoils in the genomes of eubacteria. Fluoroquinolones (FQs), successful as drugs clinically, target the enzyme to trap the gyrase-DNA complex, leading to the accumulation of double-strand breaks in the genome. Mycobacteria are less susceptible to commonly used FQs. However, an 8-methoxy-substituted FQ, moxifloxacin (MFX), is a potent antimycobacterial, and a higher susceptibility of mycobacterial gyrase to MFX has been demonstrated. Although several models explain the mechanism of FQ action and gyrase-DNA-FQ interaction, the basis for the differential susceptibility of mycobacterial gyrase to various FQs is not understood. We have addressed the basis of the differential susceptibility of the gyrase and revisited the mode of action of FQs. We demonstrate that FQs bind bothEscherichia coliandMycobacterium tuberculosisgyrases in the absence of DNA and that the addition of DNA enhances the drug binding. The FQs bind primarily to the GyrA subunit of mycobacterial gyrase, while inE. coliholoenzyme is the target. The binding of MFX to GyrA ofM. tuberculosiscorrelates with its effectiveness as a better inhibitor of the enzyme and its efficacy in cell killing.

The role of ATP in the reactions of type II DNA topoisomerases

2010 ◽  
Vol 38 (2) ◽  
pp. 438-442 ◽  
Author(s):  
Andrew D. Bates ◽  
Anthony Maxwell
Keyword(s):  
Atp Hydrolysis ◽  
Dna Gyrase ◽  
Dna Topology ◽  
Type Ii ◽  
Free Energies ◽  
Dna Topoisomerases ◽  
Type Ii Dna Topoisomerases ◽  
Topology Simplification ◽  
Hydrolysis Of

Type II DNA topoisomerases catalyse changes in DNA topology in reactions coupled to the hydrolysis of ATP. In the case of DNA gyrase, which can introduce supercoils into DNA, the requirement for free energy is clear. However, the non-supercoiling type II enzymes carry out reactions that are apparently energetically favourable, so their requirement for ATP hydrolysis is not so obvious. It has been shown that many of these enzymes (the type IIA family) can simplify the topology of their DNA substrates to a level beyond that expected at equilibrium. Although this seems to explain their usage of ATP, we show that the free energies involved in topology simplification are very small (<0.2% of that available from ATP) and we argue that topology simplification may simply be an evolutionary relic.

Structural studies of DNA gyrase

1984 ◽  
Vol 42 ◽  
pp. 164-167
Author(s):  
T. Kirchhausen ◽  
J. Wang ◽  
S. C. Harrison
Keyword(s):  
Microscopic Study ◽  
Dna Gyrase ◽  
Staphylococcal Nuclease ◽  
Dependent Manner ◽  
Type Ii ◽  
Structural Studies ◽  
Electron Microscopic ◽  
Base Pairs ◽  
Gyrase A ◽  
Negative Supercoils

DNA gyrase, a prokaryotic type II topoisomerase, can introduce negative supercoils into DNA in an ATP-dependent manner and relax negative supercoils in the absence of ATP. Gyrase is a tetramer of two A subunits (MW 105,000) and two B subunits (MW 95,000), and it interacts with about 140 base pairs of DNA (based on staphylococcal nuclease and DNase I protection experiments). We have undertaken an electron microscopic study of gyrase and its binding to DNA, in order to determine how DNA wraps in its complex with the protein.

New Strategy on Antimicrobial-resistance: Inhibitors of DNA Replication Enzymes

2019 ◽  
Vol 26 (10) ◽  
pp. 1761-1787 ◽  
Author(s):  
Lanhua Yi ◽  
Xin Lü
Keyword(s):  
Dna Replication ◽  
Antimicrobial Resistance ◽  
Enzyme Inhibitors ◽  
Dna Gyrase ◽  
Dna Helicase ◽  
Type Ii ◽  
New Strategy ◽  
Clinic Practice ◽  
Living Organisms ◽  
Replication Enzymes

Background:Antimicrobial resistance is found in all microorganisms and has become one of the biggest threats to global health. New antimicrobials with different action mechanisms are effective weapons to fight against antibiotic-resistance.Objective:This review aims to find potential drugs which can be further developed into clinic practice and provide clues for developing more effective antimicrobials.Methods:DNA replication universally exists in all living organisms and is a complicated process in which multiple enzymes are involved in. Enzymes in bacterial DNA replication of initiation and elongation phases bring abundant targets for antimicrobial development as they are conserved and indispensable. In this review, enzyme inhibitors of DNA helicase, DNA primase, topoisomerases, DNA polymerase and DNA ligase were discussed. Special attentions were paid to structures, activities and action modes of these enzyme inhibitors.Results:Among these enzymes, type II topoisomerase is the most validated target with abundant inhibitors. For type II topoisomerase inhibitors (excluding quinolones), NBTIs and benzimidazole urea derivatives are the most promising inhibitors because of their good antimicrobial activity and physicochemical properties. Simultaneously, DNA gyrase targeted drugs are particularly attractive in the treatment of tuberculosis as DNA gyrase is the sole type II topoisomerase in Mycobacterium tuberculosis. Relatively, exploitation of antimicrobial inhibitors of the other DNA replication enzymes are primeval, in which inhibitors of topo III are even blank so far.Conclusion:This review demonstrates that inhibitors of DNA replication enzymes are abundant, diverse and promising, many of which can be developed into antimicrobials to deal with antibioticresistance.

scholarly journals Inhibition of Neisseria gonorrhoeae Type II Topoisomerases by the Novel Spiropyrimidinetrione AZD0914

2015 ◽  
Vol 290 (34) ◽  
pp. 20984-20994 ◽  
Author(s):  
Gunther Kern ◽  
Tiffany Palmer ◽  
David E. Ehmann ◽  
Adam B. Shapiro ◽  
Beth Andrews ◽  
...  
Keyword(s):  
Neisseria Gonorrhoeae ◽  
Bacterial Species ◽  
Dna Gyrase ◽  
Resistant Mutant ◽  
Multidrug Resistant ◽  
Type Ii ◽  
Topoisomerase Iv ◽  
Gram Positive ◽  
Gram Negative

We characterized the inhibition of Neisseria gonorrhoeae type II topoisomerases gyrase and topoisomerase IV by AZD0914 (AZD0914 will be henceforth known as ETX0914 (Entasis Therapeutics)), a novel spiropyrimidinetrione antibacterial compound that is currently in clinical trials for treatment of drug-resistant gonorrhea. AZD0914 has potent bactericidal activity against N. gonorrhoeae, including multidrug-resistant strains and key Gram-positive, fastidious Gram-negative, atypical, and anaerobic bacterial species (Huband, M. D., Bradford, P. A., Otterson, L. G., Basrab, G. S., Giacobe, R. A., Patey, S. A., Kutschke, A. C., Johnstone, M. R., Potter, M. E., Miller, P. F., and Mueller, J. P. (2014) In Vitro Antibacterial Activity of AZD0914: A New Spiropyrimidinetrione DNA Gyrase/Topoisomerase Inhibitor with Potent Activity against Gram-positive, Fastidious Gram-negative, and Atypical Bacteria. Antimicrob. Agents Chemother. 59, 467–474). AZD0914 inhibited DNA biosynthesis preferentially to other macromolecules in Escherichia coli and induced the SOS response to DNA damage in E. coli. AZD0914 stabilized the enzyme-DNA cleaved complex for N. gonorrhoeae gyrase and topoisomerase IV. The potency of AZD0914 for inhibition of supercoiling and the stabilization of cleaved complex by N. gonorrhoeae gyrase increased in a fluoroquinolone-resistant mutant enzyme. When a mutation, conferring mild resistance to AZD0914, was present in the fluoroquinolone-resistant mutant, the potency of ciprofloxacin for inhibition of supercoiling and stabilization of cleaved complex was increased greater than 20-fold. In contrast to ciprofloxacin, religation of the cleaved DNA did not occur in the presence of AZD0914 upon removal of magnesium from the DNA-gyrase-inhibitor complex. AZD0914 had relatively low potency for inhibition of human type II topoisomerases α and β.

scholarly journals Functions of a GyrBA Fusion Protein and Its Interaction with QnrB and Quinolones

2015 ◽  
Vol 59 (11) ◽  
pp. 7124-7127 ◽  
Author(s):  
Chunhui Chen ◽  
Regis Villet ◽  
George A. Jacoby ◽  
David C. Hooper
Keyword(s):  
Escherichia Coli ◽  
Fusion Protein ◽  
Dna Gyrase ◽  
Temperature Sensitive ◽  
Type Ii ◽  
Wild Type ◽  
Interacting Protein ◽  
Content Type ◽  
Negative Supercoiling

ABSTRACTIn order to study the interactions betweenEscherichia coliDNA gyrase and the gyrase interacting protein QnrBin vivo, we constructed agyrB-gyrAfusion and validated its ability to correct the temperature-sensitive growth ofgyrAandgyrBmutants. Like wild-typegyrA, thegyrB-gyrAfusion complemented a quinolone-resistantgyrAmutant to increase susceptibility. It functioned as an active type II topoisomerase, catalyzed negative supercoiling of DNA, was inhibited by quinolone, and was protected by QnrB.

scholarly journals Insights into the Mechanism of Inhibition of Novel Bacterial Topoisomerase Inhibitors from Characterization of Resistant Mutants of Staphylococcus aureus

2015 ◽  
Vol 59 (9) ◽  
pp. 5278-5287 ◽  
Author(s):  
Sushmita D. Lahiri ◽  
Amy Kutschke ◽  
Kathy McCormack ◽  
Richard A. Alm
Keyword(s):  
Staphylococcus Aureus ◽  
Dna Gyrase ◽  
Cross Resistance ◽  
Type Ii ◽  
Topoisomerase Inhibitors ◽  
Topoisomerase Iv ◽  
Content Type ◽  
Mechanism Of Inhibition ◽  
Resistant Mutants ◽  
Dna Complex

ABSTRACTThe type II topoisomerases DNA gyrase and topoisomerase IV are clinically validated bacterial targets that catalyze the modulation of DNA topology that is vital to DNA replication, repair, and decatenation. Increasing resistance to fluoroquinolones, which trap the topoisomerase-DNA complex, has led to significant efforts in the discovery of novel inhibitors of these targets. AZ6142 is a member of the class of novel bacterial topoisomerase inhibitors (NBTIs) that utilizes a distinct mechanism to trap the protein-DNA complex. AZ6142 has very potent activity against Gram-positive organisms, includingStaphylococcus aureus,Streptococcus pneumoniae, andStreptococcus pyogenes. In this study, we determined the frequencies of resistance to AZ6142 and other representative NBTI compounds inS. aureusandS. pneumoniae. The frequencies of selection of resistant mutants at 4× the MIC were 1.7 × 10−8forS. aureusand <5.5 × 10−10forS. pneumoniae. To improve our understanding of the NBTI mechanism of inhibition, the resistantS. aureusmutants were characterized and 20 unique substitutions in the topoisomerase subunits were identified. Many of these substitutions were located outside the NBTI binding pocket and impact the susceptibility of AZ6142, resulting in a 4- to 32-fold elevation in the MIC over the wild-type parent strain. Data on cross-resistance with other NBTIs and fluoroquinolones enabled the differentiation of scaffold-specific changes from compound-specific variations. Our results suggest that AZ6142 inhibits both type II topoisomerases inS. aureusbut that DNA gyrase is the primary target. Further, the genotype of the resistant mutants suggests that domain conformations and DNA interactions may uniquely impact NBTIs compared to fluoroquinolones.

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