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Members of the genus  Acinetobacter  have been the focus recent attention due to both their clinical significance and application to molecular biology. The soil commensal bacterium  Acinetobacter baylyi  ADP1 has been proposed as a model system for molecular and genetic studies, whereas in a clinical environment,  Acinetobacter spp . are of increasing importance due to their propensity to cause serious and intractable systemic infections. Clinically, a major factor in the success of  Acinetobacter spp . as opportunistic pathogens can be attributed to their ability to rapidly evolve resistance to common antimicrobial compounds. Whole genome sequencing of clinical and environmental  Acinetobacter spp . isolates has revealed the presence of numerous multidrug transporters within the core and accessory genomes, suggesting that efflux is an important host defense response in this genus. In this work, we used the drug-susceptible organism  A. baylyi  ADP1 as a model for studies into the evolution of efflux mediated resistance in genus  Acinetobacter , due to the high level of conservation of efflux determinants across four diverse  Acinetobacter  strains, including clinical isolates. A single exposure of therapeutic concentrations of chloramphenicol to populations of  A. baylyi  ADP1 cells produced five individual colonies displaying multidrug resistance. The major facilitator superfamily pump  craA  was upregulated in one mutant strain, whereas the resistance nodulation division pump  adeJ  was upregulated in the remaining four. Within the  adeJ  upregulated population, two different levels of  adeJ  mRNA transcription were observed, suggesting at least three separate mutations were selected after single-step exposure to chloramphenicol. In the  craA  upregulated strain, a T to G substitution 12 nt upstream of the  craA  translation initiation codon was observed. Subsequent mRNA stability analyses using this strain revealed that the half-life of mutant  craA  mRNA was significantly greater than that of wild-type  craA  mRNA.

plos one

Single-Step Selection of Drug Resistant Acinetobacter baylyi ADP1 Mutants Reveals a Functional Redundancy in the Recruitment of Multidrug Efflux Systems