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. 1977 Nov;74(11):4772–4776. doi: 10.1073/pnas.74.11.4772

Nalidixic acid resistance: A second genetic character involved in DNA gyrase activity

Martin Gellert 1, Kiyoshi Mizuuchi 1, Mary H O'Dea 1, Tateo Itoh 1, Jun-Ichi Tomizawa 1
PMCID: PMC432037  PMID: 337300

Abstract

ATP-dependent DNA supercoiling catalyzed by Escherichia coli DNA gyrase was inhibited by oxolinic acid, a compound similar to but more potent than nalidixic acid and a known inhibitor of DNA replication in E. coli. The supercoiling activity of DNA gyrase purified from nalidixic acid-resistant mutant (nalAR) bacteria was resistant to oxolinic acid. Thus, the nalA locus is responsible for a second component needed for DNA gyrase activity in addition to the component determined by the previously described locus for resistance to novobiocin and coumermycin (cou). Supercoiling of λ DNA in E. coli cells was likewise inhibited by oxolinic acid, but was resistant in the nalAR mutant. The inhibition by oxolinic acid of colicin E1 plasmid DNA synthesis in a cell-free system was largely relieved by adding resistant DNA gyrase.

In the absence of ATP, DNA gyrase preparations relaxed supercoiled DNA; this activity was also inhibited by oxolinic acid, but not by novobiocin. It appears that the oxolinic acid-sensitive component of DNA gyrase is involved in the nicking-closing activity required in the supercoiling reaction. In the presence of oxolinic acid, DNA gyrase forms a complex with DNA, which can be activated by later treatment with sodium dodecyl sulfate and a protease to produce double-strand breaks in the DNA. This process has some similarities to the known properties of relaxation complexes.

Keywords: Escherichia coli, DNA supercoiling, oxolinic acid, colicin E1 DNA replication

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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