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| Status |
Public on Nov 04, 2020 |
| Title |
The small RNAs PA2952.1 and prrH as regulators of virulence, motility and iron metabolism in Pseudomonas aeruginosa |
| Organism |
Pseudomonas aeruginosa PAO1 |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Purpose: Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen that undergoes swarming motility in response to semisolid conditions with amino acids as a nitrogen source. With a genome encoding hundreds of potential intergenic small RNAs (sRNAs), P. aeruginosa can easily adapt to different conditions and stresses. We previously identified 20 sRNAs dysregulated under swarming conditions and here provide phenotypic characterization for these sRNAs.
Methods: Here, these sRNA were overexpressed in strain PAO1 and subjected to an array of phenotypic screens.
Results: Overexpression of prrH resulted in decreased swimming motility, while a ∆prrH mutant had decreased cytotoxicity and increased pyoverdine production. Overexpression of the previously uncharacterized PA2952.1 resulted in decreased swarming and swimming motility, increased gentamicin and tobramycin resistance under swarming conditions, and increased trimethoprim susceptibility. RNA-Seq and proteomics were performed on the wildtype overexpressing PA2952.1 cf. the empty vector control under swarming conditions, and revealed the differential expression of 784 genes and differential abundance of 445 proteins. Amongst these were found 82 transcriptional regulators, two-component systems, sigma and anti-sigma factors. Downstream effectors included downregulated pili, dysregulated flagellar genes, the upregulated efflux pump MexGHI-OpmD, the upregulated arn operon, and downregulated DNA synthesis genes. Genes involved in iron and zinc uptake were also dysregulated, and certain pyoverdine and phenazine genes were upregulated.
Conclusions: Overall, the sRNAs PA2952.1 and prrH appeared to be involved in regulating virulence-related programs in P. aeruginosa, including iron acquisition and motility.
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| Overall design |
Five biological replicates of the wildtype strain overexpressing PA2952.1 were compared to five biological replicates of the wildtype with the empty vector as a control.
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| Contributor(s) |
Coleman SR, Smith M, Spicer V, Lao Y, Taylor P, Mookherjee N, Hancock RE |
| Citation(s) |
33158897 |
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| Submission date |
Mar 10, 2020 |
| Last update date |
Feb 03, 2021 |
| Contact name |
Shannon Coleman |
| Organization name |
University of British Columbia
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| Street address |
2259 Lower Mall
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| City |
Vancouver |
| State/province |
BC |
| ZIP/Postal code |
V6T 1Z4 |
| Country |
Canada |
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| Platforms (1) |
| GPL18782 |
Illumina HiSeq 2500 (Pseudomonas aeruginosa PAO1) |
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| Samples (10)
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| Relations |
| BioProject |
PRJNA611835 |
| SRA |
SRP252284 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE146765_RAW.tar |
240.0 Kb |
(http)(custom) |
TAR (of TXT) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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