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Pseudomonas aeruginosa , a well - known cause of nosocomial infection, is frequently antibiotic resistant and this complicates treatment. Links between oxidative stress responses inducing antibiotic resistance through over - production of RND - type efflux pumps have been reported in  P. aeruginosa , but this has not previously been associated with MFS - type efflux pumps. Two MFS efflux pumps encoded by  mfs1  and  mfs2  were selected for study because they were found to be sodium hypochlorite (NaOCl) inducible. Antibiotic susceptibility testing was used to define the importance of these MFS pumps in antibiotic resistance and proteomics was used to characterize the resistance mechanisms involved. The results revealed that  mfs1  is NaOCl inducible whereas  mfs2  is NaOCl, N - Ethylmaleimide and t - butyl hydroperoxide inducible. Deletion of  mfs1  or  mfs2  did not affect antibiotic or paraquat susceptibility. However, over - production of Mfs1 and Mfs2 reduced susceptibility to aminoglycosides, quinolones, and paraquat. Proteomics, gene expression analysis and targeted mutagenesis showed that over - production of the MexXY RND - type efflux pump in a manner dependent upon  armZ , but not  amgRS , is the cause of reduced antibiotic susceptibility upon over - production of Mfs1 and Mfs2.  mexXY operon  expression analysis in strains carrying various lengths of  mfs1  and  mfs2  revealed that at least three transmembrane domains are necessary for  mexXY  over - expression and decreased antibiotic susceptibility. Over-expression of the MFS-type efflux pump gene  tetA (C) did not give the same effect. Changes in paraquat susceptibility were independent of  mexXY  and  armZ  suggesting that it is a substrate of Mfs1 and Mfs2. Altogether, this is the first evidence of cascade effects where the over - production of an MFS pump causes over - production of an RND pump, in this case MexXY via increased  armZ  expression.

frontiers in microbiology

Over-Expression of Hypochlorite Inducible Major Facilitator Superfamily (MFS) Pumps Reduces Antimicrobial Drug Susceptibility by Increasing the Production of MexXY Mediated by ArmZ in Pseudomonas aeruginosa