PMID:10476036
| Citation |
Hojgaard, A, Szerlong, H, Tabor, C and Kuempel, P (1999) Norfloxacin-induced DNA cleavage occurs at the dif resolvase locus in Escherichia coli and is the result of interaction with topoisomerase IV. Mol. Microbiol. 33:1027-36 |
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| Abstract |
The dif locus is a site-specific recombination site located within the terminus region of the chromosome of Escherichia coli. Recombination at dif resolves circular dimer chromosomes to monomers, and this recombination requires the XerC, XerD and FtsK proteins, as well as cell division. In order to characterize other enzymes that interact at dif, we tested whether quinolone-induced cleavage occurs at this site. Quinolone drugs, such as norfloxacin, inhibit the type 2 topoisomerases, DNA gyrase and topoisomerase IV, and can cleave DNA at sites where these enzymes interact with the chromosome. Using strains in which either DNA gyrase or topoisomerase IV, or both, were resistant to norfloxacin, we determined that specific interactions between dif and topoisomerase IV caused cleavage at that site. This interaction required XerC and XerD, but did not require the C-terminal region of FtsK or cell division. |
| Links | |
| Keywords |
Anti-Infective Agents/pharmacology; DNA Gyrase; DNA Topoisomerase IV; DNA Topoisomerases, Type II/drug effects; DNA Topoisomerases, Type II/genetics; DNA Topoisomerases, Type II/metabolism; Deoxyribonucleases, Type II Site-Specific/genetics; Deoxyribonucleases, Type II Site-Specific/metabolism; Enzyme Inhibitors/pharmacology; Escherichia coli/drug effects; Escherichia coli/genetics; Escherichia coli Proteins; Membrane Proteins/genetics; Mutation; Norfloxacin/pharmacology; Recombinases; Transposases/drug effects; Transposases/genetics; Transposases/metabolism |
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Significance
This paper showed via southern blots that Topo IV preferentially cuts DNA at the dif site in norfloxacin treated cells.
Useful Materials and Methods
Southern blots and Pulse-field electrophoresis were the more specific methods used in this paper. This paper also describes the construction of a norfloxin- resistant gyrase/parC double mutant (PK3872gyrA L83parC L80).
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