Identification of immunoreactive proteins in murine sera after immunization with outer membrane vesicles isolated from Francisella tularensis subsp. holarctica strain FSC200
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Q Exactive
ORGANISM(S): Francisella Tularensis Subsp. Holarctica Fsc200
DISEASE(S): Tularemia
SUBMITTER: 
Jana Klimentova
LAB HEAD: Jiri Stulik
PROVIDER: PXD047830 | Pride | 2024-02-22
REPOSITORIES: Pride
Similar Datasets
Project description:Francisella tularensis is a Gram-negative, facultative intracellular bacterium, causing a severe disease known as tularemia. It is known to secrete unusually shaped nanotubular outer membrane vesicles (OMV) loaded with number of virulence factors and immunoreactive proteins. In this study the nanotubes were purified from a clinical isolate of subsp. holarctica strain FSC200. We here provide a comprehensive proteomic characterization of OMV isolated from bacteria grown under different cultivation conditions simulating the diverse conditions of F. tularensis life cycle. These included conditions mimicking the environment inside the mammalian host during inflammation: oxidative stress, low pH and high temperature (42 °C); and by contrast, low temperature (25 °C) that mimicked the external milieu and agar plate cultivation. We observed several-fold increase in vesiculation rate at high temperature and low pH but the differences were not only in the amount of secreted OMV but particularly in their protein cargo. Amongst the proteins preferentially selectively packed into OMV under conditions mimicking mammalian host many were previously described as virulence factors connected to the unique intracellular trafficking of Francisella. Protein changes revealed also high probability of alterations of the bacterial surface on the level of lipopolysaccharide, polysaccharide capsule and phospholipids.
2019-09-30 | PXD013074 | Pride
Project description:Secretion of outer membrane vesicles (OMV) presents an important phenomenon in Gram-negative bacteria and they play multiple roles in their lifestyle including virulence and host-pathogen interaction. Francisella tularensis secretes unusually shaped tubular OMV. We here present a proteomic comparison of whole cell lysates, OMV and membrane fractions derived from two F. tularensis strains: a moderately virulent subsp. holarctica strain FSC200 and highly virulent subsp. tularensis SchuS4 strain. We describe a novel approach to the cross-species proteomic comparison based on creating an intersection protein database from the individual single-species databases. This approach proved to be less prone to quantification errors arising from differences in the protein sequences than using the individual databases. The consecutive comparison of the subproteomes of OMV and membranes of the two strains allowed us to distinguish the differences in protein amounts caused by global expression changes from those caused by preferential protein packing to OMV or membrane. Amongst the most different proteins we detected proteins involved in the biosynthesis and metabolism of the bacterial envelope components like O-antigen, lipid A, phospholipids, and fatty acids, as well as some of the major structural outer membrane proteins.
2021-02-08 | PXD022406 | Pride
Project description:Francisella tularensis is a Gram-negative bacteria responsible for tularemia, a disease for which the high prevalence of failure and relapses is a main concern. Directed evolution experiments carried out in presence of antibiotics revealed that acquisition of fluoroquinolones (FQ) resistance was not exclusively related to mutations in DNA gyrase genes which are the first targets of these molecules. Here, using F. tularensis LVS as model, we investigated the role of FupA/B (Fer-Utilization Protein), whose possible contribution to FQ resistance was suggested by genomic analysis of resistant isolates. Using trans-complementation/mutagenesis approaches we definitely connected FupA/B expression to FQ sensitivity. Furthermore, we showed that the virulent strain F. tularensis subsp. tularensis SCHU S4, lacking the homologous FupA protein, exhibited a lower FQ susceptibility. Importantly, the deletion of this lipoprotein promoted an increased secretion of outer membrane vesicles (OMVs). Mass spectrometry-based quantitative proteomic characterization of OMVs from LVS and LVS-∆fupA/B strains led to the identification of 801 proteins among which a subset of 23 proteins exhibited a differential abundance between OMVs from both strains and may therefore contribute to the observed increased FQ susceptibility. We finally demonstrated that OMVs are structural key elements of F. tularensis LVS biofilms providing protection against FQ. Taken together, these results showed that mutations targeting FupA/B contribute to antimicrobial resistance through quantitative and qualitative modulations of OMV biogenesis and biofilm formation, providing new basis for understanding and fighting against Francisella antibiotic resistance and/or persistence.
2019-11-12 | PXD010305 | Pride
Project description:Mass spectrometry analysis of immunoreactive protein spots revealed that seveal prostate cancer patient sera had autoantibodies to a number of proteins associated with both the glycolysis and plasminogen pathways.
2017-01-06 | PXD003968 | Pride
Project description:Mycobacterium avium subsp. paratuberculosis (MAP) is the cause of a chronic enteritis of ruminants (bovine paratuberculosis-Johne disease) that is associated with enormous worldwide economic losses for the zootechnical industries. Diagnosis is based on observation of clinical signs, on the detection of antibodies in milk or serum or on evaluation of bacterial culture from feces. The limit of these methods is that they are not able to detect the disease in the subclinical stage and are applicable only when the disease is already in an advanced status. For this reason the main purpose of this study is to use the MAP proteome to detect novel immunoreactive proteins that may be helpful for paratuberculosis diagnoses. 2D electrophoresis and 2D immunoblotting of MAP proteins were performed using sera of control cattle and paratuberculosis infected cattle in order to highlight the specific immunoreactive proteins. Among the assigned identifiers to immunoreactive spots it was found that most of them correspond to surface-located proteins while three of them have never been described before as antigens. The identification of these proteins improves scientific knowledge that could be useful for paratuberculosis diagnoses. The sequence of the identified protein can be used for the synthesis of immunoreactive peptides that could be screened for their immunoreaction against bovine sera infected with MAP.
2014-11-20 | PXD001159 | Pride
Project description:Aggregatibacter actinomycetemcomitans is an oral and systemic pathogen associated with aggressive forms of periodontitis, and with endocarditis. Outer membrane vesicles (OMVs) released by this species have been demonstrated to deliver effector proteins such as cytolethal distending toxin (CDT) and leukotoxin (LtxA) into human host cells, and to act as triggers of innate immunity upon carriage of NOD1- and NOD2-active pathogen-associated molecular patterns (PAMPs). To improve our understanding of the pathogenicity-associated functions that A. actinomycetemcomitans exports via OMVs, we have used liquid chromatography-tandem mass spectrometry (LC-MS/MS) to characterize the OMV-associated proteome of a rough-colony type clinical isolate, strain 173 (serotype e). This yielded identification of 151 proteins overlapping in three out of four independent experiments, using density gradient-purified OMVs. Further to confirming the vesicle-associated release of LtxA, and of several proteins earlier demonstrated to act as immunoreactive antigens in the human host, we identified numerous additional putative virulence-related proteins in the A. actinomycetemcomitans OMV proteome, including proteins involved in immune evasion, drug targeting, and iron/nutrient acquisition. In summary our findings are consistent with an OMV-associated proteome that exhibits several offensive and defensive functions, and they provide a comprehensive basis to further disclose roles of A. actinomycetemcomitans OMVs in periodontal and systemic disease.
2015-09-21 | PXD002509 | Pride
Project description:Francisella tularensis is a Gram-negative bacterium that causes a fatal human disease known as tularemia. The Centers for Disease Control have classified F. tularensis as Category A Tier-1 Select Agent. The virulence mechanisms of Francisella are not entirely understood. Francisella possesses very few transcription regulators, and most of these regulate the expression of genes involved in intracellular survival and virulence. The F. tularensis genome sequence analysis reveals an AraC (FTL_0689) transcriptional regulator homologous to the AraC/XylS family of transcriptional regulators. In Gram-negative bacteria, AraC activates genes required for L-arabinose utilization and catabolism. The role of the FTL_0689 regulator in F. tularensis is not known. In this study, we characterized the role of FTL_0689 in gene regulation of F. tularensis and investigated its contribution to intracellular survival and virulence. The results demonstrate that FTL_0689 in Francisella is not required for L-arabinose utilization. Instead, FTL_0689 specifically regulates the expression of the oxidative and global stress response, virulence, metabolism, and other key pathways genes required by Francisella when exposed to oxidative stress. The FTL_0689 mutant is attenuated for intramacrophage growth, and mice infected with the FTL_0689 mutant survive better than wild-type F. tularensis LVS infected mice. Based on the deletion mutant phenotype, FTL_0689 was termed osrR (oxidative stress response regulator). Altogether, this study elucidates the role of the osrR transcriptional regulator in tularemia pathogenesis.
2021-10-13 | GSE183001 | GEO
Project description:Differential expression in human peripheral blood monocytes between F. novicida-infected and uninfected, and between Francisella tularensis tularensis isolate Schu S4 and uninfected. The goal was to examine genomewide transcriptional reponses to these two strains, and identify differentially-regulated genes that may help explain the virulence of Schu S4. Keywords: Immune Response, Human Monocytes, Bacteria, Francisella
2008-08-13 | GSE12108 | GEO
Project description:Francisella tularensis may enter the body thorugh the lungs and cause fatal infection. In this study the inflammatory response to the virulent strain of Francisella (Schu4) was mapped over a 96h time-course using a custom microarray.
2011-03-01 | GSE17248 | GEO
Project description:Raghunathan2010 - Genome-scale metabolic
network of Francisella tularensis (iRS605)
This model is described in the article:
Systems approach to
investigating host-pathogen interactions in infections with the
biothreat agent Francisella. Constraints-based model of
Francisella tularensis.
Raghunathan A, Shin S, Daefler
S.
BMC Syst Biol 2010; 4: 118
Abstract:
BACKGROUND: Francisella tularensis is a prototypic example
of a pathogen for which few experimental datasets exist, but
for which copious high-throughout data are becoming available
because of its re-emerging significance as biothreat agent. The
virulence of Francisella tularensis depends on its growth
capabilities within a defined environmental niche of the host
cell. RESULTS: We reconstructed the metabolism of Francisella
as a stoichiometric matrix. This systems biology approach
demonstrated that changes in carbohydrate utilization and amino
acid metabolism play a pivotal role in growth, acid resistance,
and energy homeostasis during infection with Francisella. We
also show how varying the expression of certain metabolic genes
in different environments efficiently controls the metabolic
capacity of F. tularensis. Selective gene-expression analysis
showed modulation of sugar catabolism by switching from
oxidative metabolism (TCA cycle) in the initial stages of
infection to fatty acid oxidation and gluconeogenesis later on.
Computational analysis with constraints derived from
experimental data revealed a limited set of metabolic genes
that are operational during infection. CONCLUSIONS: This
integrated systems approach provides an important tool to
understand the pathogenesis of an ill-characterized biothreat
agent and to identify potential novel drug targets when rapid
target identification is required should such microbes be
intentionally released or become epidemic.
This model is hosted on
BioModels Database
and identified by:
MODEL1507180003.
To cite BioModels Database, please use:
BioModels Database:
An enhanced, curated and annotated resource for published
quantitative kinetic models.
To the extent possible under law, all copyright and related or
neighbouring rights to this encoded model have been dedicated to
the public domain worldwide. Please refer to
CC0
Public Domain Dedication for more information.
2015-07-30 | MODEL1507180003 | BioModels