Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

The dynamic surface proteome of allergenic fungal conidia

ABSTRACT: Fungal spores and hyphal fragments are major contributors to the allergic response in the human lung. In this study, using a trypsin-shaving-proteomics approach, we identified the surface-exposed and secreted/shed proteins of Aspergillus fumigatus conidia and investigated the dynamics of the surface proteome under different conditions, including temperature variation and germination. We find that the surface proteome of resting A. fumigatus conidia is quite dynamic, as evidenced by drastically different surface proteomes under different growth conditions. We further investigated two observed A. fumigatus surface proteins, ScwA and CweA; ScwA is specific to the Aspergillus genus and only expressed on conidia grown at higher temperatures, whereas CweA is found throughout the Aspergillus and Penicillium genera. We extended our analysis of the surface proteome of A. fumigatus to other allergy-inducing pathogens; Alternaria alternata, Penicillium rubens, and Cladosporium herbarum, and performed comparative proteomics on resting and swollen conidia, as well as secreted proteins from germinating conidia. In this dataset, we detected 125 protein orthologs in the extracellular region of all four organisms, and 42 nonorthologous proteins produced by A. fumigatus, which may be useful in the development of future diagnostics. This study highlights the dynamic nature of the conidial surface and implicates growth conditions in the antigenicity of fungal conidia.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Penicillium Rubens Neosartorya Fumigata (aspergillus Fumigatus) Alternaria Alternata Cladosporium Herbarum

SUBMITTER: Thomas Krüger

LAB HEAD: Axel A. Brakhage

PROVIDER: PXD017067 | Pride | 2020-01-24

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Similar Datasets

The role of RodAp-conserved cysteine residues in the Aspergillus fumigatus conidial surface organization

Project description:Immune inertness of Aspergillus fumigatus conidia, an airborne fungal pathogen, is attributed to its surface rodlet-layer made up of RodAp, a protein belonging to the hydrophobin family, characterized by eight conserved cysteine residues forming four disulfide bonding. Earlier, we showed that the conserved cysteine residue point (ccrp) mutations result in conidia devoid of the rodlet-layer. Here we extended our study in comparing ccrp-mutation with RODA deletion on their mutant conidial surface organization, permeability and immunoreactivity. Western blot using anti-RodAp antibodies indicated the absence of RodAp in the cytoplasm of ccrp-mutant conidia. Upon immunolabelling, ccrp-mutant conidia showed strong positivity for -(1,3)-glucan and weak positivity for -(1,3)-glucan, which was reverse in ∆rodA conidia, and the parental strain conidia were negative; all of their conidial cell wall permeability was similar. Proteomic analyses of the conidial surface exposed proteins of the ccrp-mutants, although lower in number, showed more similarities with the parental strain, but a significant difference with the RODA deletion mutant strain. Further, ccrp-mutant conidia are less immunostimulatory compared to ∆rodA conidia. Together, our data suggest that (i) the conserved cysteine residues are essential for the trafficking of RodAp and the organization of rodlet-layer on the conidial surface, and (ii) point-mutation could be an alternative strategy to study the proteins involved in the organization of fungal cell wall.

2021-02-18 | PXD020136 | Pride

Next Generation Sequencing of HUVECs stimulated by conidia of Aspergillus fumigatus

Project description:Purpose: The goal of this study is to investigate the responses of HUVECs after the stimulation of conidia of A. fumigatus Methods: HUVECs were stimulated with conidia of Aspergillus fumigatus for 2 and 6 hours. Three biological repeats of stimulated cells or un-stimulated controls were send for RNA sequencing. Results: Using an optimized data analysis workflow, we mapped about 40 million sequence reads per sample to the human genome (build hg38) and identified round 80,000 transcripts in the HUVECs upon stimulation. Conclusions: Our resutls showed the detailed analysis of HUVECs transcriptomes upton conidia of Aspergillus fumigatus stimulation.

2022-03-29 | GSE191240 | GEO

Aspergillus fumigatus melanin manipulates the cargo and kinetics of neutrophil-derived extracellular vesicles

Project description:Neutrophil-derived extracellular vesicles have regained scientific interest as potent ‘wireless’ modulators with pleiotropic effects on the immune system. A few studies have addressed their role in the context of microbial pathogenesis showing their bacteriostatic effects against Staphylococcus aureus in vitro and their potential as diagnostic markers in the onset of bacterimic sepsis in vivo. Here, we provided first insights into the vesicle release driven by the clinically relevant pathogenic fungus Aspergillus fumigatus that causes invasive infections in immunocompromised individuals worldwide often with a lethal outcome. Neutrophils, stimulated with fungal spores, potently produced vesicles that we refer as anti-fungal extracellular vesicles (afEVs). We revealed that the conidial surface pigment of A. fumigatus dihydroxynaphthalene (DHN) melanin - a well known fungal virulence factor - is a key ‘switch-on/off’ governing vesicle production. We profiled the kinetics of afEV formation and showed that DHN-melanin interferes with the vesicle number, vesicle surface maturation markers such as CD11b, CD63 and CD177 and governs the enrichment of distinct afEV subpopulations unrelated to apoptosis. Simultaneously we tracked down the fate of neutrophils during afEV formation and observed mulivesicular body formation that led to active export of CD11b, CD63 and CD177 by shedding from the surface of neutrophils onto the surface afEVs. By dissecting the proteome cargo of afEVs produced against wild-type and melanin-free conidia we verified that DHN-melanin also manipulates the type of cargo neutrophils accumulate in the vesicles upon confronting conidia with different cell wall integrity. afEVs contained higher amounts of antimicrobial peptides than spontaneously released EVs from uninfected neutrophils. Last but not least afEVs exhibit fungistatic effects and also modulate cell wall morphogenesis of hyphae.

2019-05-17 | PXD005994 | Pride

Aspergillus fumigatus whole genome transcriptomic study using AWAFUGE microarray after exposition to murine macrophages (RAW 264.7) and human lung epithelial cells (A549).

Project description:Aspergillus fumigatus is a filamentous fungus capable to cause an important disease known as invasive aspergillosis. Challenge different cell lines is a crucial strategy to undercover new virulence factors involved in the infection process. In this research, RAW 264.7 macrophages and A549 lung epithelial cells were challenge using A. fumigatus conidia in a MOI of 10 and 5 respectively. When these conidia reach 30% of germination, the total RNA was isolated (A. fumigatus alone (Control), A. fumigatus vs RAW 264.7, and A. fumigatus vs A549) and purified using the RNeasy Plant Mini Kit (Qiagen). The AWAFUGE (Agilent Whole A. fumigatus Genome Expression 44K v.1) hybridization was carried out following previously publish protocols (A-MEXP-2352 and E-MTAB-5314).

2024-01-20 | E-MTAB-13476 | biostudies-arrayexpress

Human p11-mediated re-direction of phagosomes to the recycling endosome-expulsion pathway induced by fungal pathogen

Project description:Aspergillus fumigatus invades pulmonary epithelial cells by induced phagocytosis, and survives for some time in phagosomes. Although dihydroxynaphthalene melanin on fungal conidia has been intensively investigated for its role in inhibiting phagosome maturation, a proportion of melanin-lacking mutant conidia still escaped killing by phagosomes, implying that additional mechanisms must be in place. Here, we identified the fungal surface-exposed heat shock protein HscA as an effector protein that binds the human p11 protein. A. fumigatus induces and increases the amount of p11 and anchors p11 and Annexin A2 to phagosomes and phagocytic cups, the latter facilitates phagocytosis of conidia by epithelial cells. We identified a SNP in the in the non-coding region of the human p11 gene resulting in heightened susceptibility for invasive aspergillosis in hematopoietic stem-cell transplant recipients. This finding can help for stratification of donors for hematopoietic stem cell transplantation. On phagosomes, HscA triggers an increase of p11 on the phagosomal membrane, which prevents directing the phagosome to the degradative pathway by re-directing the p11-positive phagosome to the recycling endosomal pathway. Therefore, a surface-located heat shock protein enables fungal conidia to escape phagolysosome killing.

2023-03-13 | PXD030501 | Pride

Phagolysosomal proteomics identifies DHN-melanin-regulated processes in activated macrophage in response to Aspergillus fumigatus conidia

Project description:Infection by the human pathogenic fungus Aspergillus fumigatus is initialized by the outgrowth of asexual spores (conidia) into the lung tissue of the immunocompromised host following an inhalation of the airborne conidia. The resident phagocytes, the alveolar macrophages are the first immune cells to encounter invading conidia. However, A. fumigatus conidia employ versatile mechanisms to evade the host immune defense and establish a severe invasive infection. Previously, we showed that depending on the presence of conidial 1,8-dihydroxynaphthalene (DHN) melanin, A. fumigatus circumvents intracellular killing by manipulating the phagolysosomal maturation process. Here, by comparative dual proteomics we analyzed proteins of phagolysosomes containing melanized wild-type or non-melanized pksP mutant conidia. Bioinformatics compiled a regulatory module of differentially abundant proteins that mirrors processes targeted by the fungus for immune evasion. Those are i.e. vATPAse-driven phagolysosomal acidification, endocytic trafficking, signal transduction, energy metabolism and immune response. In detail, we found alterations of vATPase complex assembly and impaired abundances of mTOR and MAPK signaling molecules, Rab5 and Vamp8 mediators of endosomal trafficking as well as Lamp1 and cathepsin Z lysosomal markers.

2018-03-08 | PXD006134 | Pride

Small RNA sequencing of Aspergillus fumigatus

Project description:Aspergillus fumigatus is an important human pathogen and a leading fungal killer. This study aimed to determine the small RNA repertoire of A. fumigatus in conidia and mycelium grown for 24 or 48 hours in liquid culture.

2023-02-14 | GSE223619 | GEO

Phagolysosomal proteomics identifies different macrophage activation profiles in response to Aspergillus fumigatus conidia

Project description:An Aspergillus fumigatus infection is initialized by the inhalation and germination of airborne asexual spores (conidia). Alveolar macrophages are the first immune cells to counteract an infection by phagocytosis of conidia and intracellular degradation of the pathogen. However, A. fumigatus circumvents its intracellular killing by the manipulation of the phagolysosomal maturation. With a comparative proteomic study of the phagolysosomal proteome of a virulent wild type A. fumigatus strain and an avirulent mutant strain we aimed at the identification of proteins and processes that are hijacked by the virulent strain. We found that wild type conidia strongly modulate the protein composition of the phagolysosome. Assembly and activity of vATPase, formation of lipid rafts, signal transduction pathways as well as the generation of energy and metabolites were significantly regulated in the wild type conidia containing phagolysosomes. In different experimental setups we confirmed a disassembly of the vATPase complex, reduced formation of lipid rafts and altered abundances of mTOR, MAPK signaling molecules, Rab5 and Vamp8 mediators of endosomal trafficking as well as LAMP1 and cathepsin Z lysosomal markers. Our interactome analysis indicates that A. fumigatus proteins target small GTPases of the endosomal trafficking system and signal transducers of the host, which might lead to a reduced fungicidal activity of the phagolysosome.

2018-03-08 | PXD005724 | Pride

Gene expression level of Aspergillus fumigatus CEA17_ΔakuBKU80 at three different stages

Project description:We report the gene expression level of Aspergillus fumigatus CEA17_ΔakuBKU80 strain in dormant conidia, in swollen conidia (after 4h of culture in Glucose 3%, Yeast Exrtract 1% liquid medium) and in germinated conidia (after 8h of culture in Glucose 3%, Yeast Extract 1% liquid medium after construction of an RNAseq library.

2016-09-26 | GSE81956 | GEO

Human monocyte-derived macrophage response to Aspergillus fumigatus

Project description:To investigate the influence of Aspergillus fumigatus on iron regulation in macrophages, we obtained macrophages in culture from human derived monocytes and co-cultured the monocyte-derived macrophages with Aspergillus conidia at a 1:1 ratio. We collected samples at 0, 2, 4, 6 and 8 hours and extracted RNA. We then performed gene expression profiling analysis using data obtained from RNA-seq of control macrophages and macrophage co-cultured with Aspergillus fumigatus at five time points.

2022-05-08 | GSE202286 | GEO

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data