Ram locus is a key regulator to trigger multidrug resistance in Enterobacter aerogenes

Alexander Molitor (1) , Chloé James (2) , Séamus Fanning (3) , Jean-Marie Pagès (1) , Anne Davin-Regli (1)

1 MCT - Membranes et cibles thérapeutiques
2 University of Salford
3 School of Agriculture

Purpose Several genetic regulators belonging to AraC family are involved in the emergence of MDR isolates of E. aerogenes due to alterations in membrane permeability. Compared with the genetic regulator Mar, RamA may be more relevant towards the emergence of antibiotic resistance. Methodology Focusing on the global regulators, Mar and Ram, we compared the amino acid sequences of the Ram repressor in 59 clinical isolates and laboratory strains of E. aerogenes. Sequence types were associated with their corresponding Multi-drug resistance phenotypes and membrane protein expression profiles using MIC and immunoblot assays. Quantitative gene expression analysis of the different regulators and their targets (porins and efflux pump components) were performed. Results In the majority of the MDR isolates tested, ramR and a region upstream of ramA were mutated but marR or marA were unchanged. Expression and cloning experiments highlighted the involvement of the ram locus in the modification of membrane permeability. Overexpression of RamA lead to decreased porin production and increased expression of efflux pump components, whereas overexpression of RamR had the opposite effects. Conclusion Mutations or deletions in ramR, leading to the overexpression of RamA predominated in clinical MDR E. aerogenes isolates andwere associated with a higher-level of expression of efflux pump components. It was hypothesised that mutations in ramR, and the self-regulating region proximal to ramA, probably altered the binding properties of the RamR repressor; thereby producing the MDR phenotype. Consequently, mutability of RamR may play a key role in predisposing E. aerogenes towards the emergence of a MDR phenotype.

Mots clés: Multidrug-Resistant (MDR); Extended-Spectrum β-Lactamase (ESBL); Minimum Inhibitory Concentration (MICs); Luria-Bertani (LB); 5-Bromo-4-chloro-3-indolyl phosphate (BCIP); nitro blue tetrazolium (NBT)

Domaines: Sciences du Vivant [q-bio]

Alexander Molitor, Chloé James, Séamus Fanning, Jean-Marie Pagès, Anne Davin-Regli. Ram locus is a key regulator to trigger multidrug resistance in Enterobacter aerogenes. Journal of Medical Microbiology, 2018, 67 (2), pp.148 - 159. ⟨10.1099/jmm.0.000667⟩. ⟨hal-01831629⟩