The Pseudomonas aeruginosa PilSR two-component system regulates both twitching and swimming motilities
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA448429 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR6927110_1.fastq.gz | Fastqsanger.gz | |||
| SRR6927110_2.fastq.gz | Fastqsanger.gz | |||
| SRR6927111_1.fastq.gz | Fastqsanger.gz | |||
| SRR6927111_2.fastq.gz | Fastqsanger.gz | |||
| SRR6927112_1.fastq.gz | Fastqsanger.gz |
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Project description:We used NGS to find that the PilS-PilR two component system of P. aeruginosa controls the expression of many non-pilus related genes and has a role in regulating swimming motility
2018-12-21 | GSE112597 | GEO
Project description:Motility is an important virulence trait for many bacterial pathogens, allowing them to position themselves in appropriate locations at appropriate times. The motility structures type IV pili and flagella are also involved in sensing surface contact, which modulates pathogenicity. In Pseudomonas aeruginosa, the PilS-PilR two-component system (TCS) regulates expression of the type IV pilus (T4P) major subunit PilA, while biosynthesis of the single polar flagellum is regulated by a hierarchical system that includes the FleSR TCS. Previous studies of Geobacter sulfurreducens and Dichelobacter nodosus implicated PilR in regulation of non-T4P-related genes, including some involved in flagellar biosynthesis. Here we used transcriptome sequencing (RNA-seq) analysis to identify genes in addition to pilA with changes in expression in the absence of pilR Among the genes identified were 10 genes whose transcription increased in the pilA mutant but decreased in the pilR mutant, despite both mutants lacking T4P and pilus-related phenotypes. The products of these inversely dysregulated genes, many of which were hypothetical, may be important for virulence and surface-associated behaviors, as mutants had altered swarming motility, biofilm formation, type VI secretion system expression, and pathogenicity in a nematode model. Further, the PilSR TCS positively regulated transcription of fleSR, and thus many genes in the FleSR regulon. As a result, pilSR deletion mutants had defects in swimming motility that were independent of the loss of PilA. Together, these data suggest that in addition to controlling T4P expression, PilSR could have a broader role in the regulation of P. aeruginosa motility and surface sensing behaviors.IMPORTANCE Surface appendages such as type IV pili and flagella are important for establishing surface attachment and infection in a host in response to appropriate cues. The PilSR regulatory system that controls type IV pilus expression in Pseudomonas aeruginosa has an established role in expression of the major pilin PilA. Here we provide evidence supporting a new role for PilSR in regulating flagellum-dependent swimming motility in addition to pilus-dependent twitching motility. Further, even though both pilA and pilR mutants lack PilA and pili, we identified sets of genes downregulated in the pilR mutant and upregulated in a pilA mutant as well as genes downregulated only in a pilR mutant, independent of pilus expression. This finding suggests that change in the inner membrane levels of PilA is only one of the cues to which PilR responds to modulate gene expression. Identification of PilR as a regulator of multiple motility pathways may make it an interesting therapeutic target for antivirulence compounds.
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Project description:The ability of Pseudomonas aeruginosa to detect host immune responses and coordinate the production of virulence factors is crucial for its high pathogenicity. However, the regulatory mechanisms underlying this process remain largely unknown. In this study, we extensively characterize the conserved two-component system CprRS in P. aeruginosa and uncover the role of the sensor histidine kinase CprS as a receptor for the human host defense peptide LL-37, thereby modulating bacterial virulence. In the presence of LL-37, CprS functions as a phosphatase, targeting the intracellular activator CprR for phosphorylation at position D53, ultimately leading to CprR activation. By employing quantitative proteomics and transcription analysis, we further elucidate that CprR induction under LL-37 treatment promotes the expression of type III secretion system effectors. This, in turn, impedes the expression of proinflammatory cytokines and significantly increases bacterial cytotoxicity towards macrophages. Notably, the mutation of either cprS or cprR markedly diminishes bacterial survival in both macrophage and insect infection models. Collectively, our study uncovers a novel regulatory mechanism of the two-component system in P. aeruginosa, which enables the bacterium to sense and respond to human innate immune system responses while ensuring a timely balance of virulence genes.
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Project description:Pseudomonas aeruginosa is a common nosocomial pathogen which produces siderophores to solubilize and transport chelated Fe3+ to aid its survival in both the environment and the host. However, there is a lack of comprehensive understanding regarding the molecular mechanisms underlying siderophore synthesis, uptake, and regulation within various ecological niches. In this study, we demonstrated that the BfmRS two-component system, part of the core genome of P. aeruginosa, plays a crucial role in siderophore metabolism. We have identified BfmS as an osmosensing histidine kinase that responds to external osmolytes, then modulates the activation of the response regulator BfmR. Under high osmolality, BfmR could directly bind to the promoters of pvd, fpv, and femARI gene clusters, thereby enhancing their expression and promoting siderophore metabolism. The proteomic and phenotypic analyses confirmed that deletion of bfmRS results in reduced expression levels of siderophore-related proteins as well as siderophore production. Importantly, loss of bfmR or bfmS significantly impaired bacterial survival in both iron deficiency medium and mouse lung infection models. Furthermore, phylogenetic analysis revealed that BfmRS is highly conserved and widely distributed across Pseudomonas species, evidences also proved that the BfmR of P. putida KT2440 and P. sp. MRSN12121 activated siderophore genes in response to high osmolality. Overall, this study sheds light on the previously unexplored signal transduction pathway, BfmRS, which governs the siderophore regulation in Pseudomonas species through perceiving an osmotic upshift. Considering that siderophores serve as unique social mediators, our findings contribute to a better understanding of how siderophores facilitate bacterial interactions with their eukaryotic hosts and contribute to the establishment of stable communities.
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Project description:Two component systems (TCSs) control a large proportion of virulence factors in Pseudomonas aeruginosa. Yet, investigations on inhibitors of regulatory pathways of TCS remain scare, despite their potential in anti-virulence strategies. This work encompasses the working mechanism of PIT4, a protein derived from the lytic P. aeruginosa phage LSL4. This viral protein inhibits bacterial motility and in particular twitching motility, while reducing the virulence of P. aeruginosa towards HeLa cells. Via differential gene expression and a yeast two-hybrid screen, PIT4 was shown to interact with components of different two component systems. In one-on-one interaction assays, it was confirmed that PIT4 interacts with the histidine kinases FleS, PilS and PA2882, through interaction with the histidine kinase domain. As such, this work highlights the potential of previously unknown phage proteins in virulence regulation of multidrug resistant pathogens that might be exploited for anti-virulence strategies and biotechnological applications.
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Project description:We report the transcriptome of Pseudomonas aeruginosa PA14 under swarming conditions as compared to a swimming control
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Project description:The widespread presence of antibiotic-resistant bacteria in the environment has been recognized as an important emerging environmental contaminant. Hospital wards, as a special public indoor environment, are of great concern for the risks associated with this emerging environmental contaminant. Pseudomonas aeruginosa, a common nosocomial bacterium, is a contamination risk in the hospital environment due to its drug resistance and transmission of virulence factors. Notably, the antimicrobial peptide-sensing two-component system (TCS) ParRS and CprRS have been implicated in dynorphin-induced signaling, but the underlying Manuscript2 mechanism has remained elusive. In this study, we performed proteomic analysis to systematically investigate the contributions of ParRS and CprRS to P. aeruginosa pathogenesis and dynorphin-induced resistance to polymyxins. Additionally, we characterized the significance of the extracellular sensor domains of ParS and CprS in dynorphin perception. Furthermore, through structural biology, we identified additional TCS sensors with similar extracellular domain conformations, which also directly interacted with dynorphin in vitro. This suggests convergent evolution in different bacterial TCSs for host-derived synthetic peptide signal transmitting. Our findings establish a link between CAMPs resistance associated TCSs and virulence regulation of common nosocomial bacteria. This further illustrates the danger of this emerging contaminant for the environment and humans.
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Project description:Microarray were done on 4 independent cultures. PA01 was grown for 6 hours to late log growth phase (OD600 0.9-1.0) in liquid BM2-swarming media under shaking conditions(swimming) or for 18 hours at 37C on BM2-swarming plate containing 0.5% (w/v) agar and 0.1% casamino acids. Cells were harvested either by centrifugation(swimming) or by a sterile inoculating loop (swarming) and were resuspended n BM2-swarming media supplement with RNAprotect reagent (Qiagen)
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Project description:Gene expression in a Geobacter sulfurreducens strain overexpressing the two-component system GsuTCS1
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