Pseudomonas aeruginosa mexR and mexEF Antibiotic Efflux Pump Variants Exhibit Increased Virulence
Abstract
:1. Introduction
2. Results
2.1. Deletion of mexAB Restores Aztreonam Susceptibility of Evolved P. aeruginosa
2.2. Deletion of mexAB Does Not Affect Virulence of Evolved P. aeruginosa
2.3. Genome Sequencing Reveals a Previously Undetected 19 kb Deletion in PAO1-AzEvB8
2.4. Mutation of mexEF Increases Swarming While mexR Mutation Does not
2.5. Deletion of either mexR or mexEF Increases P. aeruginosa Virulence
2.6. Efflux Pump Inhibition Increases Rhamnolipid Virulence Factor Expression
3. Discussion
3.1. Deletion of mexEF and Overexpression of mexAB through mexR Mutation Can Each Increase Virulence of P. aeruginosa
3.2. Understanding Why rhlA Was Not Expressed in the PAO1-AzEvB8 Strain That Exhibited Greater Swarming Than WT
3.3. Relationship of These Findings to Previous Research
3.4. Differences among Infection Models Help Explain Why Previous Studies Concluded Different Effects of Efflux Pump Inhibition on Virulence
3.5. Relevance of mexEF Mutations in P. aeruginosa Clinical Isolates
3.6. Implications for CF and the Use of Efflux Pump Inhibitor as Potential Therapies
3.7. Limitations of the Present Work
4. Materials and Methods
4.1. Bacterial Strains and Growth Conditions
4.2. Deletion Plasmid Construction
4.3. Transformation of P. aeruginosa Deletion Mutants
4.4. PAO1 Transformation with rhlA Reporter Plasmid
4.5. DNA Extraction, Purification, and PCR
4.6. Swarming Assay
4.7. Gradient Diffusion Antibiotic Susceptibility Testing
4.8. rhlA GFP Reporter Assay
4.9. Murine Lung Infection
4.10. Genome Sequencing and Analysis
4.11. Statistics
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Vaillancourt, M.; Limsuwannarot, S.P.; Bresee, C.; Poopalarajah, R.; Jorth, P. Pseudomonas aeruginosa mexR and mexEF Antibiotic Efflux Pump Variants Exhibit Increased Virulence. Antibiotics 2021, 10, 1164. https://doi.org/10.3390/antibiotics10101164
Vaillancourt M, Limsuwannarot SP, Bresee C, Poopalarajah R, Jorth P. Pseudomonas aeruginosa mexR and mexEF Antibiotic Efflux Pump Variants Exhibit Increased Virulence. Antibiotics. 2021; 10(10):1164. https://doi.org/10.3390/antibiotics10101164
Chicago/Turabian StyleVaillancourt, Mylene, Sam P. Limsuwannarot, Catherine Bresee, Rahgavi Poopalarajah, and Peter Jorth. 2021. "Pseudomonas aeruginosa mexR and mexEF Antibiotic Efflux Pump Variants Exhibit Increased Virulence" Antibiotics 10, no. 10: 1164. https://doi.org/10.3390/antibiotics10101164