Discoordinate gene expression of gyrA and gyrB in response to DNA gyrase inhibition in Escherichia coli.
The intracellular level of DNA supercoiling is regulated in Escherichia coli by a homeostatic control mechanism that includes DNA gyrase and topoisomerase I gene expression. Despite several biochemical and genetical evidence that supports the existence of a homeostatic regulation mechanism, there are only few studies focusing gyrA and gyrB gene expression in connection to the mechanism involved in the regulation of DNA supercoiling in vivo. To study DNA gyrase gene expression and to be able to isolate mutants with altered expression of DNA gyrase, we constructed a new chromosomal reporter system based on two translational fusions of gyrA and gyrB to lacZ Using this stable monitor system in a robust wild type, we simultaneously studied the influence of several inhibitors of DNA gyrase (quinolones and coumarins) on gyrA and gyrB gene expression as well as on the intracellular level of DNA supercoiling. Surprisingly, we found a delayed and differential response of gyrA and gyrB gene expression following inhibition of DNA gyrase by quinolones or coumarins. Whereas both groups of drugs were able to increase the expression of gyrA, the gyrB gene expression was only induced by the coumarins. Although the action of the quinolones was able to alter DNA supercoiling, we never observed any induction of gyrB from the chromosome. These results revealed that the gene expressio of gyrA appears to be more sensitive to alterations in DNA supercoiling than the gyrB gene expression and suggest that probably additional regulatory mechanisms on the post-translational level might be involved in the regulation of DNA supercoiling and DNA gyrase gene expression.
Neumann
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Institut für, Martin-Luther-Universität, Domplatz, Germany.
Quiñones
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A
D049933:Aminocoumarins; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003374:Coumarins / Q000494:pharmacology; D027081:DNA Gyrase; D004250:DNA Topoisomerases, Type II / Q000235:genetics* / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D004278:DNA, Superhelical / Q000032:analysis / Q000378:metabolism*; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D007763:Lac Operon; D009268:Nalidixic Acid / Q000494:pharmacology; D009675:Novobiocin / Q000494:pharmacology; D010093:Oxolinic Acid / Q000494:pharmacology; D010957:Plasmids; D014176:Protein Biosynthesis; D015363:Quinolones / Q000494:pharmacology; D059005:Topoisomerase II Inhibitors; D001616:beta-Galactosidase / Q000378:metabolism
D049933:Aminocoumarins; D003374:Coumarins; D004269:DNA, Bacterial; D004278:DNA, Superhelical; D015363:Quinolones; D059005:Topoisomerase II Inhibitors; D009675:Novobiocin; D009268:Nalidixic Acid; D001616:beta-Galactosidase; D027081:DNA Gyrase; D004250:DNA Topoisomerases, Type II; D010093:Oxolinic Acid; C004628:coumermycin