Drug susceptibility of Burkholderia pseudomallei multidrug transporters assessed by an agar-based assay
Data files
Abstract
The Gram-negative pathogen Burkholderia pseudomallei possesses multiple resistance-nodulation-division superfamily transporters that contribute to multidrug resistance, including BpeB and BpeF. Structural studies of BpeB and BpeF have identified a hydrophilic patch in their substrate-binding pocket. To investigate the relationship between this hydrophilic patch and substrate specificity, mutant analyses were performed using an Escherichia coli recombinant expression system. Drug susceptibility tests of BpeB and BpeF mutants showed up to a 64-fold increase in susceptibility compared with the wild type. Growth curve analyses revealed that BpeB mutants exhibited increased resistance to aminoglycosides, which are not transported by the wild type. These findings suggest that the hydrophilic patches in the substrate-binding pockets of BpeB and BpeF are involved in the substrate specificity.
Dataset DOI: 10.5061/dryad.bnzs7h4qz
Dataset overview
This dataset contains photographic images of agar plates obtained from drug susceptibility assays using Escherichia coli strains heterologously expressing multidrug transporters derived from Burkholderia pseudomallei.
The images document bacterial growth in the presence of various drugs and compounds and were used to evaluate the contribution of the expressed transporters to drug susceptibility phenotypes.
Experimental context
Drug susceptibility agar plate assays were performed to assess the effects of Burkholderia-derived multidrug transporters on bacterial drug susceptibility. Recombinant E. coli strains expressing individual transporters were plated on agar media containing different drugs or compounds, and bacterial growth was documented by photography. These assays were performed using both wild-type transporters and multiple mutant variants to assess their effects on substrate specificity.
The dataset consists of image files documenting bacterial growth across drug concentration series. In addition, a table summarizing the minimum inhibitory concentration (MIC) values determined from these assays is provided as a separate file (MIC.xlsx).
Description of Data Files
File: AmrB.zip
Description: Image files from the drug susceptibility agar plate assays for AmrB
File: BpeB.zip
Description: Image files from the drug susceptibility agar plate assays for BpeB
File: BpeF.zip
Description: Image files from the drug susceptibility agar plate assays for BpeF
File: MIC.xlsx
Description: Table summarizing the MIC values determined from the agar plate assays
Dataset organization
The dataset is organized hierarchically by protein and drug. The top-level folders correspond to three multidrug transporters, AmrB, BpeB, and BpeF.
Within each protein folder, subfolders are organized by drug. A total of 14 drugs and compounds were evaluated, and each drug-specific subfolder contains photographs of agar plates obtained from assays performed with that drug.
File description
Each image file is a photograph of an agar plate showing bacterial growth in the presence of a specific drug or compound. Within each drug-specific subfolder, the images correspond to agar plates containing different concentrations of the indicated drug. Drug susceptibility assays were performed using a two-fold serial dilution series.
The numbers following the drug abbreviations written on the plates indicate the drug concentration. A value of “1” corresponds to a concentration of 100 µg/mL, and each subsequent increase in number represents a two-fold decrease in concentration (e.g., “2” indicates 50 µg/mL, and “3” indicates 25 µg/mL). For DDM and UDM, the concentrations are ten-fold higher than those of other drugs; in these cases, “1” corresponds to 1000 µg/mL.
No image processing was applied to the files.
Plate layout
In each agar plate image, the top two rows contain strains expressing the following transporters, from left to right:
- pBAD24 (empty vector)
- BpeB (wild type)
- BpeB_D407N+D408N (transport-inactive mutant, negative control)
- BpeB_S132A
- BpeB_S132A+S133A
- BpeB_S132A+S133A+S134D
- BpeB_T176V
The middle two rows contain strains expressing the following transporters, from left to right:
- pBAD24 (empty vector)
- BpeF (wild type)
- BpeF_D410N+D411N (transport-inactive mutant, negative control)
- BpeF_S134A
- BpeF_S134A+S135A
- BpeF_S134A+S135A+P136D
- BpeF_Q178V
- BpeF_S134A+S135A+Q178V
The bottom two rows contain strains expressing the following transporters, from left to right:
- BpeB_S132A+S133A+S134D+T176E
- BpeB_S132A+S133A+S134D+S135N+F136A+L137Q
- BpeB_S167R
- pBAD24 (empty vector)
- AmrB (wild type)
- AmrB_D406N+D407N (transport-inactive mutant, negative control)
MIC data
The file MIC.xlsx contains a table summarizing the MIC values determined from the agar plate assays. MIC values were determined based on the presence or absence of colony growth observed in the corresponding agar plate images.
MIC values reported as “> 4000” indicate that no growth inhibition was observed at the highest concentration evaluated.
Drug abbreviations
The abbreviations used for drugs and compounds in folder names are as follows:
- ACR, acriflavine
- CHL, chloramphenicol
- CLI, clindamycin
- DDM, n-dodecyl-β-D-maltoside
- EB, ethidium bromide
- ERY, erythromycin
- GEN, gentamicin
- KAN, kanamycin
- LVX, levofloxacin
- NOR, norfloxacin
- TET, tetracycline
- TMP, trimethoprim
- TOB, tobramycin
- UDM, n-undecyl-β-D-maltoside
Related publication
This dataset supports the findings reported in the following publication:
Title: Substrate specificity of Burkholderia pseudomallei multidrug transporters is influenced by the hydrophilic patch in the substrate-binding pocket
Journal: FEBS Letters (2025)
DOI: https://doi.org/10.1002/1873-3468.70248
The MIC values provided in MIC.xlsx correspond to those reported in the related publication. A summary of the strains used and the basic experimental procedures is provided above. For additional methodological details including strain construction, expression conditions, and full assay protocols, please refer to the related publication.
Contact information
For questions regarding this dataset, please contact:
Corresponding author: Ui Okada
Affiliation: Institute of Science Tokyo
E-mail: uiokada@life.isct.ac.jp