Exposure to diverse antimicrobials induces the expression of qnrB1, qnrD and smaqnr genes by SOS-dependent regulation.
OBJECTIVE
Direct SOS-dependent regulation of qnrB genes by fluoroquinolones mediated by LexA was reported. The smaqnr gene, on the Serratia marcescens chromosome, and qnrD both contain a putative LexA box. The aim of this study was to evaluate whether smaqnr or qnrD genes are induced via SOS-dependent mechanisms, and to investigate whether other antimicrobial agents induce qnrB, qnrD and smaqnr expression.


METHODS
RT-PCR was used to evaluate qnrB1, qnrD and smaqnr expression. Different concentrations of ciprofloxacin, levofloxacin, moxifloxacin and ceftazidime were evaluated as inducers. Additionally, the promoter regions of qnrB1, qnrD and smaqnr were fused transcriptionally to green fluorescent protein and used in reporter gene assays. Disc diffusion assays with different antimicrobial agents were used to detect induction. Measurements of transcriptional induction by ciprofloxacin were carried out using a plate reader.


RESULTS
RT-PCR assays showed that qnrB1, qnrD and smaqnr were induced at different concentrations of ciprofloxacin, moxifloxacin, levofloxacin and ceftazidime, increasing transcription 1.5- to 16.3-fold compared with basal expression, and depending on the antimicrobial agent and promoter analysed. The reporter gene assays showed that the qnrB1, qnrD and smaqnr genes were induced by ciprofloxacin, as expected, but also by ceftazidime, ampicillin and trimethoprim in Escherichia coli wild-type strains, but not in the recA-deficient E. coli HB101. Induction was not evident for imipenem or gentamicin.


CONCLUSIONS
β-Lactams and trimethoprim, along with fluoroquinolones, induce transcription of qnrB, qnrD and smaqnr genes using SOS-dependent regulation. These results show the direct SOS-dependent regulation of a low-level fluoroquinolone resistance mechanism in response to other antimicrobials.
Departmento de Microbiología,  Universidad de Sevilla, Seville,  Spain.
A
A
Briales
J M
JM
Rodriguez-Martinez
C
C
Velasco
J
J
Machuca
P
P
Díaz de Alba
J
J
Blazquez
A
A
Pascual
D000900:Anti-Bacterial Agents / Q000378:metabolism*; D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000096:biosynthesis*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D024841:Fluoroquinolones / Q000378:metabolism*; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D008826:Microbial Sensitivity Tests; D011401:Promoter Regions, Genetic; D060888:Real-Time Polymerase Chain Reaction; D013014:SOS Response, Genetics; D012706:Serratia marcescens / Q000187:drug effects* / Q000235:genetics*; D014295:Trimethoprim / Q000378:metabolism; D047090:beta-Lactams / Q000378:metabolism
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D024841:Fluoroquinolones; D047090:beta-Lactams; D049452:Green Fluorescent Proteins; D014295:Trimethoprim