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Phenotypical adaptation of Pseudomonas aeruginosa in the presence of siderophore-antibiotic conjugates during epithelial cell infection

ABSTRACT: This study aims at investigating the ability of Pseudomonas aeruginosa to detect the presence of siderophore-antibiotic conjugates in an epithelial cell infection assay. We show that the presence of siderophore-antibiotic conjugates induces the transcription and expression of their corresponding transporters, indicating the bacteria are able to sense the chelators in their environment and adapt their phenotype accordingly.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Homo Sapiens (human) Pseudomonas Aeruginosa Pao1

TISSUE(S): Epithelial Cell, Cell Culture

SUBMITTER: Paola Cantero

LAB HEAD: Laurence Sabatier

PROVIDER: PXD020378 | Pride | 2021-02-18

REPOSITORIES: Pride

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Phenotypic adaptation of Pseudomonas aeruginosa by hacking siderophores produced by other microorganisms

Project description:This study aims at investigating the ability of Pseudomonas aeruginosa to detect the presence of exosiderophores in an epithelial cell infection assay. We show that the presence of exosiderophores induces the transcription and expression of their corresponding transporters, and that bacteria sense the presence of the chelators in their environment and adapt their phenotype to access iron via the exosiderophore present

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Gene expression profiling of siderophore-drug resistant mutant of Pseudomonas aeruginosa PAO1

Project description:The aim of the experiment was to identify genes differentially expressed between the susceptible wild type strain P. aeruginosa PAO1 (PT5) and a mutant resistant to a drug-siderophore conjugate, in order to obtain information on the resistance mechanism(s). A mutant of PT5 able to grow at 4 mg/l BAL30072, a drug-siderophore conjugate, was selected in vitro . The susceptible wild type strain PT5 and the mutant (BAL6) were grown in LB medium and the mutant also in the presence of 4 mg/l BAL30072 to mid-exponential growth phase (OD600 =2) in triplicate cultures. RNA was extracted using the RNeasy Kit (Qiagen). A total of nine Affymterix P. aeruginosa arrays were hybridized (one for each replicate) under standard conditions.

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Project description:Iron is an essential nutrient for the opportunistic pathogen Pseudomonas aeruginosa, as for almost all living organisms. To access this element, the pathogen is able to express at least 15 different iron-uptake pathways, the vast majority involving small iron chelators called siderophores. Indeed, P. aeruginosa produces two siderophores, pyoverdine and pyochelin, but can also use many produced by other microorganisms. This implies that the bacterium expresses appropriate TonB-dependent transporters (TBDTs) at the outer membrane to import the ferric form of each of the siderophores used. These transporters are highly selective for a given ferri-siderophore complex or for siderophores with similar chemical structures. Here, we show that P. aeruginosa can also use rhizoferrin, staphyloferrin A, aerobactin, and schizokinen as siderophores to access iron. Growth assays in iron-restricted conditions and 55Fe uptake assays showed that the two alpha-carboxylate type siderophores rhizoferrin-Fe and staphyloferrin A-Fe are transported into P. aeruginosa cells by the TBDT ActA (PA3268). Among the mixed alpha-carboxylate/hydroxamate type siderophores, we found aerobactin-Fe to be transported by ChtA (as previously described) and schizokinen-Fe by ChtA and another unidentified TBDT.

2023-09-21 | PXD043056 | Pride

The Serratia marcescens siderophore, serratiochelin, is necessary for full virulence during bloodstream infection

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RNA-seq analysis in simulated microgravity unveils down-regulation of the beta-rhizobial siderophore phymabactin

Project description:Exploiting the symbiotic interaction between crops and nitrogen-fixing bacteria is a simple and ecological solution to promote plants growth in prospective extraterrestrial human outposts. In this study, we investigated the adaptation of the legume symbiont Paraburkholderia phymatum to simulated microgravity (s0-g) at the transcriptome level by performing an RNA-Seq analysis. The results revealed a drastic effect on gene expression, with roughly 23 % of P. phymatum genes being differentially regulated in s0-g. Among those, 951 genes were upregulated and 858 downregulated in the cells grown in s0-g compared to terrestrial gravity (1g). Genes involved in posttranslational modification, proteins turnover and chaperones production were upregulated in s0-g, while those involved in translation, ribosomal structure and biosynthesis, motility or inorganic ions transport were downregulated. Specifically, the whole phm gene cluster, previously predicted to be involved in the production of a hypothetical siderophore, phymabactin, was approximatively 20-fold downregulated in microgravity. Accordingly, a phm-gfp reporter strain showed less expression in s0-g and iron uptake was reduced in microgravity. By constructing a mutant strain (ΔphmJK) we confirmed that the phm gene cluster codes for the only siderophore secreted by P. phymatum. In fact, in contrast to P. phymatum wild-type, ΔphmJK did not produce any siderophores on chrome azurol S plates. These results not only provide a deeper understanding of the physiology of symbiotic organisms exposed to space-like conditions, but also increase our understanding of iron acquisition in rhizobia.

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The proteomic response of a marine Vibrio to low iron and desferrioxamine siderophore bioavailability

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2021-04-19 | PXD011427 | Pride

Siderophore piracy as a resilience factor for Bacteroidetes during enteric pathogen infection

Project description:Siderophore piracy as a resilience factor for Bacteroidetes during enteric pathogen infection

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The two component system BfmRS controls siderophore production in Pseudomonas aeruginosa

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.

2023-10-30 | PXD046481 | iProX

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