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Mass spectrometry-based characterization of secretomes from different strains of Pseudomonas chlororaphis and Pseudomonas entomophila.

ABSTRACT: To characterize the molecular basis of cytotoxicity of different Pseudomonas species and strains, we analyzed the protein content of secretomes of three P. chlororaphis strains (CIP63, CIP75 and the reference strain PA23) and of six P. entomophila strains (L48 WT, or deleted for various virulence factors: the global activator GacA, the pore-forming toxin Mnl, the pore-forming toxin ExlA, and double mutants for these genes).

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Pseudomonas Entomophila L48 Pseudomonas Chlororaphis

SUBMITTER: Yohann Couté

LAB HEAD: Yohann Couté

PROVIDER: PXD029397 | Pride | 2022-07-25

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	P.chlororaphis.mzid.gz	Mzid
	P.entomophila.mzid.gz	Mzid
	Pc_CIP63-R1.mgf	Mgf
	Pc_CIP63-R1.raw	Raw
	Pc_CIP63-R2.mgf	Mgf

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Publications

Genomic erosion and horizontal gene transfer shape functional differences of the ExlA toxin in Pseudomonas spp.

Job Viviana V Gomez-Valero Laura L Renier Adèle A Rusniok Christophe C Bouillot Stephanie S Chenal-Francisque Viviane V Gueguen Erwan E Adrait Annie A Robert-Genthon Mylène M Jeannot Katy K Panchev Peter P Elsen Sylvie S Fauvarque Marie-Odile MO Couté Yohann Y Buchrieser Carmen C Attrée Ina I

iScience 20220614 7

Two-partner secretion (TPS) is widespread in the bacterial world. The pore-forming TPS toxin ExlA of Pseudomonas aeruginosa is conserved in pathogenic and environmental Pseudomonas. While P. chlororaphis and P. entomophila displayed ExlA-dependent killing, P. putida did not cause damage to eukaryotic cells. ExlA proteins interacted with epithelial cell membranes; however, only ExlA Pch induced the cleavage of the adhesive molecule E-cadherin. ...[more]

PMID: 35789842

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Pseudomonas aeruginosa MipA-MipB envelope proteins are new sensors of polymyxin

Project description:Due to the rising incidence of antibiotic resistant infections, last-line antibiotics polymyxins have resurged in the clinics together with the appearance of new bacterial strategies of escape. The Gram-negative opportunistic pathogen Pseudomonas aeruginosa develops resistance to colistin/polymyxin by distinct molecular mechanisms, mostly involving the modification of the lipid A component of the LPS by proteins encoded within the arnBCDATEF-ugD (called arn) operon. We characterized a polymyxin-induced operon, named mipBA, present in P. aeruginosa group of strains devoid of the arn operon. Mass spectrometry-based quantitative proteomics showed that the absence of MipBA modifies the membrane proteome, notably impacting ParRS-regulated proteins, in response to polymyxin.

2024-01-12 | PXD043253 | Pride

Pseudomonas aeruginosa CLJ3 genome sequencing, transcriptomics, and proteomics [transcriptomics]

Project description:Gene expression profiles of two Pseudomonas aeruginosa taxonomic outlier clinical isolates, CLJ1 and CLJ3 [CLJ3] Pseudomonas aeruginosa taxonomic outliers emerged recently as infectious for humans, provoking hemorrhagic pneumonia. Those bacteria lack classical type III secretion system, and utilize the pore-forming toxin for infection. Two clones CLJ1 and CLJ3 belonging to these taxonomic outliers have been isolated from the same patient at two different times during hospitalization. P. aeruginosa CLJ3 displays antibiotic resistance phenotype, while CLJ1 is more cytotoxic on epithelial and endothelial cells.

2018-11-30 | GSE123107 | GEO

Pseudomonas aeruginosa CLJ1 genome sequencing, transcriptomics, and proteomics [transcriptomics]

Project description:Gene expression profiles of two Pseudomonas aeruginosa taxonomic outlier clinical isolates, CLJ1 and CLJ3 [CLJ1] Pseudomonas aeruginosa taxonomic outliers emerged recently as infectious for humans, provoking hemorrhagic pneumonia. Those bacteria lack classical type III secretion system, and utilize the pore-forming toxin for infection. Two clones CLJ1 and CLJ3 belonging to these taxonomic outliers have been isolated from the same patient at two different times during hospitalization. P. aeruginosa CLJ3 displays antibiotic resistance phenotype, while CLJ1 is more cytotoxic on epithelial and endothelial cells.

2018-11-30 | GSE123106 | GEO

Comparison of S. aureus PVL-positive and PVL-negative isogenic strains in exponential and stationary phase of growth

Project description:The Staphylococcus aureus Panton Valentine leukocidin (PVL) is a pore-forming toxin secreted by strains epidemiologically associated with the current outbreak of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) and with the often lethal necrotizing pneumonia. To investigate the role of PVL in pulmonary disease, we tested the pathogenicity of clinical isolates, isogenic PVL-negative and PVL-positive S. aureus strains, as well as purified PVL, in a mouse acute pneumonia model. Here we show that PVL is sufficient to cause pneumonia and that the expression of this leukotoxin induces global changes in transcriptional levels of genes encoding secreted and cell-wall-anchored staphylococcal proteins, including the lung inflammatory factor staphylococcal protein A (Spa). Keywords: comparative transcription profile in the presence or absence of PVL toxin

2007-02-13 | GSE6985 | GEO

Role of RccR in the transcriptional control of primary carbon metabolism in Pseudomonas

Project description:To characterise the RccR regulon, we performed a ChIP-seq assay using a polyclonal RccR antiserum on SBW25 WT/rccR strains grown in minimal pyruvate and glycerol media. We were able to identify 8 RccR binding sites from this experiment including one in the rccR promoter region itself. The peaks identified in our RccR ChIP-seq assays corresponded to strongly enriched regions relative to the respective rccR controls. All 8 peaks were localized in intergenic regions, upstream of one or more genes.

2017-08-24 | E-MTAB-5745 | biostudies-arrayexpress

Analysis of the Hfq regulon in Pseudomonas fluorescens SBW25.

Project description:Hfq is a transcriptional and translational pleiotropic regulator in several bacteria. RNA-Seq, Ribo-Seq and Proteomic analyses were carried out in the wild-type and a hfq deletion strain of Pseudomonas fluorescens SBW25 with the intention to separate the influence of Hfq on the transcript stability and translation. This submission relates to the RNA-Seq data only. RNA was extracted from two replicate cultures each of SBW25-WT and SBW25-Δhfq strains and, after removal of ribosomal RNA, subjected to RNA-Seq in an Illumina NextSeq500 machine. The resulting sequence data was analysed by mapping to the reference sequence of Pseudomonas fluorescens SBW25 as available in the Genbank accession NC_012660.

2019-02-07 | E-MTAB-5396 | biostudies-arrayexpress

Vibrio vulnificus MARTX toxin effector domains differentially remodel the transcriptional response of intestinal epithelial cells

Project description:Vibrio vulnificus is an foodborne pathogen that can cause gastroenteritis and septicemia in humans. V. vulnificus secretes a multifunctional autoprocessing repeats-in-toxin (MARTX) toxin as an essential virulence factor to cause disease. MARTX toxins are pore-forming toxins that translocate multiple functionally independent effector domains into a target cell. MARTX toxins of V. vulnificus can contain anywhere from 3 to 5 of the 10 identified effector domains and strains with different effector repertories having varying virulence potential. The goal of this study was to compare how different effector combinations from an F-type MARTX toxin differentially remodel the transcriptional response of human intestinal epithelial cells (IECs). F-type MARTX toxins contain five effector domains – the actin crosslinking domain (ACD), two copies the makes caterpillar floppy-like domain (MCF), and alpha-beta hydrolase (ABH) domain, and the Ras/Rap1 specific endopeptidase (RRSP). Cultured human IECs were treated with V. vulnificus or strains modified to secrete a toxin with only ACD, ACD with MCF-ABH, ACD with RRSP, or no active effectors. We demonstrate that when no active effectors are present, the bacterium induces minimal changes in the transcriptional profile of IECs. However, the strains containing different effector combinations each uniquely remodeled the transcriptional profile of IECs. These data provide insight into how V. vulnificus strains with varying effector combinations can differentially regulate the host cell response to cause disease.

2021-11-21 | GSE189194 | GEO

Outer membrane vesicle of Pseudomonas aerugionsa

Project description:Objectives: This study aimed to investigate the microbiological characteristics of outer membrane vesicles (OMVs) derived from Pseudomonas aeruginosa (P. aeruginosa) to understand their mechanisms of inhibition of Acinetobacter baumannii (A. baumannii) in vitro. Methods: We assessed the inhibitory effects of P. aeruginosa on A. baumannii using a modified cross-streak assay. Subsequently, OMVs were extracted from P. aeruginosa strains using high-speed centrifugation, tangential flow filtration, ultrafiltration, and ultracentrifugation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transmission electron microscopy (TEM), and nanoparticle tracking assays (NTAs) were performed to confirm the presence of the extracted OMVs. P. aeruginosa-derived OMVs’ inhibitory activity against A. baumannii was tested using a modified time-kill assay. The proteomic analysis of OMVs revealed potential antibacterial protein clusters with diverse functions. Results: P. aeruginosa 022 (PA022) demonstrated inhibition of A. baumannii in the cross-streak assay. The protein levels of OMVs for PA022 and P. aeruginosa ATCC 27853 (PA ATCC 27853) were 1665 and 428.6 μg/mL, respectively. Additionally, PA 022 and PA ATCC 27853 exhibited variable patterns and sizes in the SDS-PAGE, TEM, and NTA. Furthermore, the growth inhibitory effect of PA022 OMVs on A. baumannii was evaluated using a modified time-kill assay. The proteomic analysis of OMVs revealed potential antibacterial protein clusters in PA 022 associated with virulence, motility, and post-translational modifications. Conclusion: Our study contributes to the understanding of P. aeruginosa OMVs characteristics and their inhibitory effect against A. baumannii, providing insights into the development of alternative therapeutic approaches against multi-drug resistance.

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Ribosome profiling of Pseudomonas fluorescens SBW25 and its hfq deletion mutant.

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