Modification of norfloxacin inhibition of DNA gyrase induced by a 28 KDal DNA binding protein.

We have previously studied a clinical isolate of Providencia stuartii which showed high levels of resistance to 4-quinolones, aminoglycoside and beta-lactam antibiotics (Landini et al., 1987). DNA gyrase from this isolate was inhibited for 50\% of activity at a concentration of 15 microM of norfloxacin, which is about 5-fold higher compared to the 50\% inhibitory concentration for a standard DNA gyrase. It has been described that 4-quinolone inhibition of DNA gyrase is caused by their binding to DNA and by the distortion induced in DNA tertiary structure, and that affinity binding of 4-quinolones is different for DNAs in different structures. In order to detect whether the interaction between pAT 153 and a protein able to modify DNA tertiary structure could affect norfloxacin inhibitory concentrations for DNA gyrase we purified from the clinical isolate of Providencia stuartii a DNA binding protein of about 28 KDal which induces changes in supercoiling degree of DNA. Assays of DNA gyrase activity were performed on the complex pAT 153-DNA binding protein-norfloxacin. Results showed an increase from 15 microM to 20 microM of 50\% norfloxacin inhibitory concentration for DNA gyrase when pAT 153 was complexed with the 28 KDal protein.