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| Status |
Public on Mar 15, 2024 |
| Title |
A multiplexed chemical screen identifies a novel, species-specific Pseudomonas aeruginosa inhibitor that targets LPS interaction with the outer membrane protein, OprH [1401_dataset2] |
| Organism |
Pseudomonas aeruginosa UCBPP-PA14 |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
The surge of antimicrobial resistance in recent decades threatens efficacy of current antibiotics, particularly against Pseudomonas aeruginosa, a highly resistant gram-negative pathogen. The asymmetric outer membrane of P. aeruginosa combined with its array of efflux pumps provide a barrier to xenobiotic intracellular accumulation, thus making the discovery of novel drugs with whole cell antibacterial activity very challenging. We adapted PROSPECT, a genome-wide, target-based, whole cell screening strategy, to take a focused approach to discover small molecule probes with specific activity against engineered P. aeruginosa mutants depleted for essential proteins localized at the outer membrane. We identified BRD1401, a small molecule that has specific activity against a P. aeruginosa mutant depleted for the essential lipoprotein, OprL. Genetic studies identified a novel link between OprL and the non-essential, outer membrane β barrel protein, OprH, to modulate BRD1401 activity. BRD1401 directly bound to OprH to disrupt the known interaction between OprH and lipopolysaccharide (LPS) in vitro and in whole bacteria. OprH also biochemically interacted with OprL, thus providing a link between outer membrane and peptidoglycan in P. aeruginosa. Thus, a whole cell, multiplexed screen against P. aeruginosa identified a species-specific inhibitor and probe molecule that revealed novel pathogen biology.
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| Overall design |
P. aeruginosa strains, either wildtype PA14 strains, or engineered oprL-hypomorph or ΔoprL or ΔPA14_50630 strains, were exposed to either 0.5% DMSO vehicle control or 256 µM BRD1401 in standard LB medium for 120 minutes.
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| Contributor(s) |
Hung DT, Poulsen BE, Warrier T, Romano KP, Bagnall JS |
| Citation missing |
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| Submission date |
Jan 24, 2024 |
| Last update date |
Mar 15, 2024 |
| Contact name |
Thulasi Warrier |
| E-mail(s) |
twarrier@broadinstitute.org
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| Organization name |
Broad Institute
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| Department |
IDMP
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| Lab |
Hung Lab
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| Street address |
415 Main Street
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| City |
Cambridge |
| State/province |
MA |
| ZIP/Postal code |
02142 |
| Country |
USA |
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| Platforms (1) |
| GPL27892 |
Illumina NovaSeq 6000 (Pseudomonas aeruginosa UCBPP-PA14) |
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| Samples (30)
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| Relations |
| BioProject |
PRJNA1068580 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE254108_1401_dataset2_counts_final_012224.csv.gz |
295.9 Kb |
(ftp)(http) |
CSV |
SRA Run Selector |
| Raw data are available in SRA |
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