Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Phagosome proteome of mock and Atg8 silenced Entamoeba histolytica strains

ABSTRACT: The enteric protozoan parasite Entamoeba histolytica have high phagocytic ability. Phagocytosis is also important for the pathogenicity of this parasite; molecular mechanisms of phagocytosis and phagosome maturation is focused. Atg8 is well studied autophagy marker protein. We previously shown that E. histolytica Atg8 translocate to nascent trogosomes and expression silencing of the Atg8 caused retardation of phagosome acidification. To investigate how Atg8 regulates phagosome maturation, here we conducted proteome analysis of phagosomes isolated from an E. histolytica strain in which atg8 gene expression are silenced (atg8 gene silenced, atg8-gs) and its mock control strain (transfected with psAP2-Gunma).

INSTRUMENT(S): Orbitrap Fusion ETD

ORGANISM(S): Entamoeba Histolytica Hm-1:imss-a

TISSUE(S): Diploid Cell

DISEASE(S): Amebiasis

SUBMITTER: Kumiko Tsukui

LAB HEAD: Kumiko Nakada-Tsukui

PROVIDER: PXD034376 | Pride | 2022-10-20

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Similar Datasets

Pacific oyster phagosome LC-MSMS

Project description:Phagosomes are task-force organelles of innate immune systems responsible for the recognition, processing, and ultimately destruction of invading microorganisms. Among invertebrate and vertebrate species, evolutionary diversity and continuity abound in the protein machinery executing this coordinately regulated process, though its dynamics and full scope of regulators remain incompletely understood. In order to clarify molecular mechanisms underlying phagocytosis, we studied phagocyte response to beads and Vibrio species, using hemocytes of the Pacific oysters (Crassostrea gigas) as a marine invertebrate model. Phagosomes from different stages of phagocytosis were isolated by density-gradient centrifugation, and more than 400 phagosome-associated proteins were subsequently identified via high-throughput quantitative proteomics. In modeling key networks of phagosomal protein-protein interactions, our results support the essential roles of several processes driving phagosome formation and maturation, including actin cytoskeleton remodeling, and signal transduction by myosin and Rab proteins. In further quantitative proteomic analysis, trends of both upregulation and downregulation of protein expression were observed proteins during phagosome formation and maturation. Notably, the signal transducers CgRhoGDI and CgPI4K were implicated. In addition, six Rab proteins including Rab1, Rab2, Rab7, Rab11, RaB21, and Rab33 were also identified as active regulators or mediators in hemocyte phagocytosis. Our current work illustrates the diversity and dynamic interplay of phagosomal proteins, providing a framework for better understanding host-microbe interactions during phagosome formation and maturation in under-examined invertebrate species.

2020-04-27 | PXD015810 | Pride

The E3 ubiquitin ligase RNF115 regulates phagosome maturation and host response to infection

Project description:Phagocytosis is an essential process for the microbicidal function of professional phagocytes, in which phagosome maturation plays a critical role. Macrophage activation by interferons (IFN) increases their microbicidal activity, but delays phagosomal maturation. Here, we investigated the role of ubiquitylation in phagosome functions. We show that phagosomal proteins are ubiquitylated and that phagosomes contain diverse ubiquitin chain types, including atypical ubiquitin chains. IFN-γ activation of macrophages substantially enhanced phagosomal ubiquitylation of both innate immune response proteins and vesicle trafficking proteins. We identified the E3 ubiquitin ligase RNF115, which is enriched on phagosomes of IFN-γ activated macrophages, as an important regulator of phagosomal maturation. Loss of RNF115 protein or ubiquitin ligase activity facilitated enhanced phagosomal maturation, and increased cytokine responses to Staphylococcus aureus. Our data suggests that both innate immune signalling and phagolysosomal trafficking are controlled through ubiquitylation. In conclusion, we identify RNF115 and ubiquitylation as important regulators of innate immune signalling from the phagosome during bacterial infections.

2022-07-20 | PXD026843 | Pride

Spatiotemporal changes of the phagosomal proteome in dendritic cells in response to LPS stimulation

Project description:Dendritic cells (DCs) are professional phagocytes that use innate sensing and phagocytosis to internalize and degrade self as well as foreign material, such as pathogenic bacteria, within phagosomes. These intracellular compartments are equipped to generate antigenic peptides that serve as source for antigen presentation to T cells initiating adaptive immune responses. Nonetheless, the phagosomal proteome of DCs is only partially studied, in particular in response to inflammatory cues. The phagosomal proteome is highly dynamic as it changes during phagosome maturation, when phagosomes sequentially interact with endosomes and lysosomes. In addition, the activation status of the phagocyte can modulate the phagosomal composition and is able to shape phagosomal functions. In this study, we determined spatiotemporal changes of the proteome of DC phagosomes during their maturation and compared resting and lipopolysaccharide (LPS)-stimulated bone marrow-derived DCs by label-free, quantitative mass spectrometry. Ovalbumin-coupled latex beads were used as phagocytosis model system and revealed that LPS-treated DCs show decreased recruitment of proteins involved in phagosome maturation, such as subunits of the vacuolar proton ATPase, cathepsin B, D, S and RAB7. In contrast, those phagosomes were characterized by an increased recruitment of proteins involved in antigen cross-presentation, e.g. different subunits of the proteasome and MHC I molecules, tapasin etc., confirming increased antigen presentation efficacy in those cells. In addition, we identified several phagosomal proteins that were not previously associated with phagosomal functions and reveal a novel role for immune-responsive gene 1 (IRG1) in phagocytic uptake of particles in resting DCs.

2019-02-28 | PXD012155 | Pride

LRRK2 signalling pathway in Mycobacterium tuberculosis

Project description:LRRK2 promotes Mycobacterium tuberculosis clearance by regulating phagosome maturation in mouse macrophages

2018-05-16 | PXD006909 | Pride

Temporal and ligand effects on phagosome maturation

Project description:Systems biology analysis of temporally-resolved phagosome proteomes following uptake via key phagocytic receptors Macrophages operate at the forefront of innate immunity, and their discrimination of pathogenic versus “self” particles is critical for a number of responses including efficient pathogen killing, antigen presentation and cytokine induction. In order to efficiently phagocytose and destroy the particles, macrophages express an array of receptors to sense and internalise prey particles. In this manuscript, particles conjugated to seven ligands representing pathogen-associated molecular patterns, immune opsonins or apoptotic cell markers were inoculated to murine bone marrow-derived macrophages (BMDMs) and proteomes of isolated phagosomes were accurately quantified among conditions and across a timecourse. While we identified a clear functional differentiation over the three timepoints, only subtle differences were detected between certain ligand-phagosomes, suggesting that triggering of receptors through a single ligand has only mild effects on phagosome proteome and function. This data shows for the first time a systematic time-course analysis of BMDM phagosomes and how phagosome fate is regulated by the receptors triggered for phagocytosis.

2015-09-10 | PXD000772 | Pride

Effect of Rom1 silencing on gene expression in Entamoeba histolytica

Project description:Entamoeba histolytica trophozoites epigenetically silenced for ameobapore A (G3 parasites) were additionally silenced for the Rhomboid gene Rom1 (EHI_197460) using the mechanism outlined in Mirelman et al. Parasite. 2008 Sep;15(3):266-74 (PMID 18814693). Rom1 silencing was confirmed by RT-PCR. Gene expression in these parasites was compared to that of the parent G3 strain. RNA was isolated from axenically grown G3 and Rom1 silenced parasites, and hybridized to a custom Affymetrix array for E. histolytica. Two biological replicates were performed for each condition.

2010-07-07 | E-GEOD-21522 | biostudies-arrayexpress

Expression data form EhCPBF1 gene silenced strain.

Project description:Enteric protozoan parasite Entamoeba histolytica is a causative agent of amebiasis. One of the major virulence factors of this parasite is cysteine protease (CP). To understand the molecular basis of intracellular trafficking of CP, we identified CP5 binding protein and named cysteine protease binding protein family 1 (CPBF1). To study the function of CPBF1, we generate CPBF1 gene silenced strain and analyzed the expression profile by custom made microarray. E. histolytica G3 strain was transfected with pSAP2 vector or pSAP2-CPBF1 and established the transformats by selecting the transfected cells with G418. After confirm the gene silencing of CPBF1 by RT-PCR, we performed the transcriptome analysis of pSAP2 (mock) and pSAP2-CPBF1 (gene silenced) cells.

2011-12-13 | E-GEOD-34368 | biostudies-arrayexpress

Effect of Rom1 silencing on gene expression in Entamoeba histolytica

2010-07-07 | GSE21522 | GEO

Rab7D small GTPase is involved in phago-, trogocytosis and cytoskeletal reorganization in the enteric protozoan Entamoeba histolytica

Project description:To know the role of Rab7 isotypes in enteric protozoan parasite Entamoeba histolytica, one out of nine Rab7 isotypes, Rab7D, was specifically silenced and examined the effect on virulence-related phenotypes. To clarify transcriptomic difference caused by rab7d gene silencing, RNA-seq analysis was conducted.

2020-09-01 | GSE152875 | GEO

Expression data form EhCPBF1 gene silenced strain.

2011-12-13 | GSE34368 | 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