Dataset Information


Interactions between QnrB, QnrB mutants, and DNA gyrase.

ABSTRACT: Plasmid-encoded protein QnrB1 protects DNA gyrase from ciprofloxacin inhibition. Using a bacterial two-hybrid system, we evaluated the physical interactions between wild-type and mutant QnrB1, the GyrA and GyrB gyrase subunits, and a GyrBA fusion protein. The interaction of QnrB1 with GyrB and GyrBA was approximately 10-fold higher than that with GyrA, suggesting that domains of GyrB are important for stabilizing QnrB1 interaction with the holoenzyme. Sub-MICs of ciprofloxacin or nalidixic acid reduced the interactions between QnrB1 and GyrA or GyrBA but produced no reduction in the interaction with GyrB or a quinolone-resistant GyrA:S83L (GyrA with S83L substitution) mutant, suggesting that quinolones and QnrB1 compete for binding to gyrase. Of QnrB1 mutants that reduced quinolone resistance, deletions in the C or N terminus of QnrB1 resulted in a marked decrease in interactions with GyrA but limited or no effect on interactions with GyrB and an intermediate effect on interactions with GyrBA. While deletion of loop B and both loops moderately reduced the interaction signal with GyrA, deletion of loop A resulted in only a small reduction in the interaction with GyrB. The loop A deletion also caused a substantial reduction in interaction with GyrBA, with little effect of loop B and dual-loop deletions. Single-amino-acid loop mutations had little effect on physical interactions except for a ?105I mutant. Therefore, loops A and B may play key roles in the proper positioning of QnrB1 rather than as determinants of the physical interaction of QnrB1 with gyrase.

PROVIDER: S-EPMC4538543 | BioStudies |

REPOSITORIES: biostudies

Similar Datasets

| S-EPMC4007428 | BioStudies
| S-EPMC3147658 | BioStudies
| S-EPMC2493125 | BioStudies
| S-EPMC4997844 | BioStudies
| S-EPMC6462232 | BioStudies
| S-EPMC89495 | BioStudies
| S-EPMC3137097 | BioStudies
| S-EPMC4356753 | BioStudies
| S-EPMC2292561 | BioStudies
| S-EPMC5075108 | BioStudies