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BatR: A novel regulator of antibiotic tolerance in P. aeruginosa biofilms

ABSTRACT: Pseudomonas aeruginosa is a multidrug-resistant opportunistic human pathogen. Chronic infections are associated with biofilms, conferring resistance to antimicrobial agents and complicating treatment strategies. This study focuses on understanding the role of the uncharacterized gene PA3049, upregulated under biofilm conditions. In the context of P. aeruginosa biofilms, PA3049 emerged as a player in withstanding antimicrobial challenges both in vitro and in clinically validated infection models. Under antibiotic conditions, the deletion of PA3049 resulted in reduced pyocyanin production and altered abundance of enzymes controlling denitrification, pyoverdine, and hydrogen cyanide biosynthesis. Notably, PA3049 directly interacts with two kinases implicated in antibiotic tolerance, inactivating their active sites. Renamed as the Biofilm antibiotic tolerance Regulator (BatR), PA3049 is a key player in P. aeruginosa biofilm maintenance and antimicrobial tolerance. These findings contribute to understanding the complex bacterial lifestyle in biofilms, shedding light on a previously uncharacterized gene with significant implications for combating multidrug-resistant infections.

INSTRUMENT(S):

ORGANISM(S): Pseudomonas Aeruginosa Pao1

TISSUE(S): Cell Culture

SUBMITTER: Gerhard Saalbach

LAB HEAD: Jacob Malone

PROVIDER: PXD050997 | Pride | 2026-06-26

REPOSITORIES: Pride

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Similar Datasets

BatR: A novel regulator of antibiotic tolerance in P. aeruginosa biofilms

Project description:Pseudomonas aeruginosa is a multidrug-resistant opportunistic human pathogen. Chronic infections are associated with biofilms, conferring resistance to antimicrobial agents and complicating treatment strategies. This study focuses on understanding the role of the uncharacterized gene PA3049, upregulated under biofilm conditions. In the context of P. aeruginosa biofilms, PA3049 emerged as a player in withstanding antimicrobial challenges both in vitro and in clinically validated infection models. Under antibiotic conditions, the deletion of PA3049 resulted in reduced pyocyanin production and altered abundance of enzymes controlling denitrification, pyoverdine, and hydrogen cyanide biosynthesis. Notably, PA3049 directly interacts with two kinases implicated in antibiotic tolerance, inactivating their active sites. Renamed as the Biofilm antibiotic tolerance Regulator (BatR), PA3049 is a key player in P. aeruginosa biofilm maintenance and antimicrobial tolerance. These findings contribute to understanding the complex bacterial lifestyle in biofilms, shedding light on a previously uncharacterized gene with significant implications for combating multidrug-resistant infections.

2026-06-26 | PXD050995 | Pride

BatR: A novel regulator of sub-inhibitory antibiotic tolerance in Pseudomonas aeruginosa biofilms

Project description:Pseudomonas aeruginosa is a multidrug-resistant opportunistic pathogen, with chronic infections often associated with biofilms that enhance antibiotic resistance. This study investigates the uncharacterized gene PA3049, which is upregulated under biofilm conditions, to determine its role in infection, biofilm formation, and antimicrobial tolerance. o Using bioinformatics, infection models, and molecular microbiology, we determined that PA3049 contributed to biofilm establishment both in vitro and in high-validity infection models. We also identified its role in bacterial survival under sub-inhibitory concentrations of antibiotics and its impact on pyocyanin production. Proteomic analysis revealed that PA3049 upregulates the R2-type pyocin cluster, which drives explosive cell lysis and extracellular DNA (eDNA) release during early stages of P. aeruginosa biofilm development. Additionally, PA3049 interacts with PA0486, an uncharacterized Ser/Thr protein kinase implicated in pyocyanin production and bacterial killing, suggesting a putative in vivo mechanism of action. o Renamed as the Biofilm antibiotic tolerance Regulator (BatR), PA3049 emerges as a key player in P. aeruginosa biofilm maintenance and resistance. These findings provide new insights into bacterial biofilm dynamics and highlight two previously uncharacterized genes with potential implications for combating multidrug-resistant infections

2026-06-26 | PXD062857 | Pride

Experimental evolution of Pseudomonas aeruginosa biofilms exposed to furanone C-30 with and without tobramycin

Project description:The aim of this study is to evaluate the evolutionary robustness of the quorum sensing inhibitor (QSI) furanone C-30 for the treatment of P. aeruginosa biofilm infections. We repeatedly exposed P. aeruginosa biofilms to furanone C-30 (with or without tobramycin) in the synthetic cystic fibrosis sputum medium (SCFM2) and characterized the genotype and phenotype of the evolved lineages. P. aeruginosa biofilms were grown in SCFM2 for 24 h after which the treatment in fresh SCFM2 was added to obtain a final concentration of 20 µg/ml tobramycin and 100 µg/ml furanone C-30. The negative control was treated with fresh SCFM2, including the same amount of DMSO (0.25%) as for the biofilms treated with C-30. After 24 h of static incubation at 37°C, biofilms were sonicated and vortexed in order to disintegrate the biofilm aggregates. After each cycle the number of CFU was determined and an aliquot of the culture was stored at -80°C in Microbank vials to allow further tests on the evolved strains. A sample from the treated biofilm was used to prepare a new overnight culture, in order to start a new cycle. For each treatment three independent lineages were established, that were each exposed for 16 cycles. Whole-genome sequencing was performed on the wild type P. aeruginosa PAO1 and on the exposed lineages after 5, 10 and 16 cycles.

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In vitro evolution of Pseudomonas aeruginosa AA2 biofilms in the presence of cystic fibrosis lung microbiome members

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2024-01-22 | PXD043651 | Pride

Fitness Landscape of Antibiotic Tolerance in Pseudomonas aeruginosa Biofilms

Project description:Bacteria in biofilms have higher antibiotic tolerance than their planktonic counterparts. A major outstanding question is the degree to which the biofilm-specific cellular state and its constituent genetic determinants contribute to this hyper-tolerant phenotype. Here, using genome-wide functional profiling of a complex, heterogeneous mutant population of Pseudomonas aeruginosa MPAO1, we identified large sets of mutations that contribute to antibiotic tolerance predominantly in the biofilm or planktonic setting only. Our mixed population-based experimental design recapitulated the complexity of natural biofilms and, unlike previous studies, revealed clinically observed behaviors including the emergence of quorum sensing-deficient mutants. Our study revealed a substantial contribution of the cellular state to the antibiotic tolerance of biofilms, providing a rational foundation for the development of novel therapeutics against P. aeruginosa biofilm-associated infections. This dataset compares the expression of SAH108, a strain with enhanced antibiotic tolerance in the biofilm state, to expression in wild-type strains. We compared the expression of two biological replicates from strain SAH108 to samples from three wild-type, reference strains. All samples were collected from exponentially-growing planktonic cultures.

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Identification of genes important for response of Pseudomonas aeruginosa biofilms to ciprofloxacin exposure using TraDIS-Xpress

Project description:Pseudomonas aeruginosa is an opportunistic pathogen that can cause severe infections in immunocompromised individuals, such as patients with cystic fibrosis where it commonly forms biofilms. Ciprofloxacin is used extensively to treat P. aeruginosa infections, but its effectiveness can be significantly reduced due to biofilm formation. Although many individual genes associated with biofilm formation have been characterised, the genetic basis of P. aeruginosa biofilm fitness related to antibiotics challenge remain unexplored. In this study we employed a high-density TraDIS-Xpress library of P. aeruginosa PAO1 to assay the impact of gene disruptions or altered gene expression on biofilm formation at different concentrations of ciprofloxacin. Gene fitness was analysed by comparing the biofilm samples to planktonic samples harvested at 12h, 24h and 48h with and without ciprofloxacin. Gene determinants of survival for biofilms at different stages of maturity in the presence and absence of ciprofloxacin were identified.

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Standardization of in vitro method of Biofilm formation by Clinical Isolates of Pseudomonas aeruginosa and elucidating its Biofilm-specific Proteins through LC-Orbitrap HRMS-based Peptidomics

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