Transcriptome of Histomonas meleagridis virulent and attenuated strains
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ABSTRACT:
PROVIDER: PRJEB19109 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| ERR1810606.fastq.gz | Fastqsanger.gz | |||
| ERR1810607.fastq.gz | Fastqsanger.gz |
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Project description:The current study aimed to detect and identify significant differentially expressed proteins between a virulent and an attenuated Histomonas meleagridis strain which was in vitro co-cultivated with Escherichia coli DH5α. Two-dimensional gel electrophoresis (2-DE) was used for proteome visualization , gel image software for computational detection of significantly up-regulated protein spots and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS) for protein identification. The statistical analysis fulfilling two criteria (> or = 3-fold up-regulation and P<0.05) detected 119 differentially expressed protein spots out of which 62 spots were located in gels of the virulent strain and 57 spots in gels of the attenuated strain. The mass spectrometric analysis of 32 spots, up-regulated in gels of the virulent strain, showed that they are of H. meleagridis origin. As opposed to this, the mass spectrometric analysis of 49 protein spots , up-regulated in the gels of the attenuated strain , identified 32 spots as specific to the protozoan. Additionally, the analysis identified a number of E. coli DH5α proteins which were detected as differentially expressed by the computational gel image and statistical analysis.
2017-12-07 | PXD007579 | Pride
Project description:Histomonas meleagridis Genome sequencing and assembly
| PRJNA594289 | ENA
Project description:The current work aimed to detect and identify significantly differentially expressed proteins between a virulent and an attenuated Histomonas meleagridis strain which were in vitro co-cultivated with Escherichia coli DH5alpha. This was done by applying 2D-DIGE and sequential window acquisition of all theoretical spectra (SWATH) MS as tools on the two well-defined strains. In the gel-based experiments, 49 identified protein spots were found to be differentially expressed, of which 37 belonged to the in vitro cultivated virulent strain and 12 to the attenuated one. The most frequently identified proteins in the virulent strain take part in cytoskeleton formation, carbohydrate metabolism and adaptation to stress. However, post-translationally modified or truncated ubiquitous cellular proteins such as actin and GAPDH were identified as upregulated in multiple gel positions. This indicated their contribution to processes not related to cytoskeleton and carbohydrate metabolism, such as fibronectin or plasminogen binding. On the other hand, proteins involved in cytoskeleton, cell division and cytoskeleton organization were frequently observed in the attenuated strain. The findings of the gel-based studies were supplemented by the gel-free SWATH MS analysis, which identified and quantified 42 significantly differentially regulated proteins. In this case proteins with peptidase activity, metabolic proteins and actin-regulating proteins were the most frequent findings in the virulent strain, while proteins involved in hydrogenosomal carbohydrate metabolism dominated the results in the attenuated one.
2018-11-21 | PXD009319 | Pride
Project description:The current work aimed to detect and identify significant differentially expressed proteins between a virulent and an attenuated Histomonas meleagridis strain which was in vitro co-cultivated with Escherichia coli DH5alpha. This was done by applying 2D-DIGE and sequential window acquisition of all theoretical spectra (SWATH) MS as tools on the two well-defined strains. In the gel-based experiments, 49 identified protein spots were found to be differentially expressed, of which 37 belonged to the in vitro cultivated virulent strain and 12 to the attenuated one. The most frequently identified proteins in the virulent strain take part in cytoskeleton formation, carbohydrate metabolism and adaptation to stress. However, post-translationally modified or truncated ubiquitous cellular proteins such as actin and GAPDH were identified as upregulated in multiple gel positions. This indicated their contribution to processes not related to cytoskeleton and carbohydrate metabolism, such as fibronectin or plasminogen binding. On the other hand, proteins involved in cytoskeleton, cell division and cytoskeleton organization were frequently observed in the attenuated strain. The findings of the gel-based studies were supplemented by the gel-free SWATH MS analysis, which identified and quantified 42 significantly differentially regulated proteins. In this case proteins with peptidase activity, metabolic proteins and actin-regulating proteins were the most frequent findings in the virulent strain, while proteins involved in hydrogenosomal carbohydrate metabolism dominated the results in the attenuated one.
2018-11-22 | PXD009332 | Pride
Project description:The current work aimed to detect and identify significant differentially expressed proteins between a virulent and an attenuated Histomonas meleagridis strain which were in vitro co-cultivated with Escherichia coli DH5alpha. This was done by applying 2D-DIGE and sequential window acquisition of all theoretical spectra (SWATH) MS as tools on the two well-defined strains. In the gel-based experiments, 49 identified protein spots were found to be differentially expressed, of which 37 belonged to the in vitro cultivated virulent strain and 12 to the attenuated one. The most frequently identified proteins in the virulent strain take part in cytoskeleton formation, carbohydrate metabolism and adaptation to stress. However, post-translationally modified or truncated ubiquitous cellular proteins such as actin and GAPDH were identified as upregulated in multiple gel positions. This indicated their contribution to processes not related to cytoskeleton and carbohydrate metabolism, such as fibronectin or plasminogen binding. On the other hand, proteins involved in cytoskeleton, cell division and cytoskeleton organization were frequently observed in the attenuated strain. The findings of the gel-based studies were supplemented by the gel-free SWATH MS analysis, which identified and quantified 42 significantly differentially regulated proteins. In this case proteins with peptidase activity, metabolic proteins and actin-regulating proteins were the most frequent findings in the virulent strain, while proteins involved in hydrogenosomal carbohydrate metabolism dominated the results in the attenuated one.
2018-12-10 | PXD009316 | Pride
Project description:The current work aimed to detect and identify significantly differentially expressed proteins between a virulent and an attenuated Histomonas meleagridis strain which were in vitro co-cultivated with Escherichia coli DH5alpha. This was done by applying 2D-DIGE and sequential window acquisition of all theoretical spectra (SWATH) MS as tools on the two well-defined strains. In the gel-based experiments, 49 identified protein spots were found to be differentially expressed, of which 37 belonged to the in vitro cultivated virulent strain and 12 to the attenuated one. The most frequently identified proteins in the virulent strain take part in cytoskeleton formation, carbohydrate metabolism and adaptation to stress. However, post-translationally modified or truncated ubiquitous cellular proteins such as actin and GAPDH were identified as upregulated in multiple gel positions. This indicated their contribution to processes not related to cytoskeleton and carbohydrate metabolism, such as fibronectin or plasminogen binding. On the other hand, proteins involved in cytoskeleton, cell division and cytoskeleton organization were frequently observed in the attenuated strain. The findings of the gel-based studies were supplemented by the gel-free SWATH MS analysis, which identified and quantified 42 significantly differentially regulated proteins. In this case proteins with peptidase activity, metabolic proteins and actin-regulating proteins were the most frequent findings in the virulent strain, while proteins involved in hydrogenosomal carbohydrate metabolism dominated the results in the attenuated one.
2023-03-11 | PXD034844 | Pride
Project description:Background: Histomonas meleagridis is an anaerobic, intercellular parasite that infects the Galliformes such as turkeys and chickens. In recent years, the reemergence of Histomoniasis has caused serious economic losses as drugs to treat the disease have been banned. At present, studies on H. meleagridis mainly focus on virulence, gene expression analysis, and the innate immunity of the host. However, there are no studies on differential expression of miRNAs (DEMs) in host immune and inflammatory response induced by H. meleagridis infection in chickens. In this study, the expression profile of cecum miRNA at 10 and 15 days post-infection (DPI) with Chinese JSYZ-F strain H. meleagridis was studied by high-throughput sequencing. Results: Compared with the control group, 94 and 127 DEMs were found in the cecum of infected chickens at 10 DPI (CE vs CC) and 15 DPI (CEH vs CCH), respectively, of which 60 DEMs were shared at two-time points. Gene Ontology (GO) enrichment analysis of the target genes of DEMs showed that 881 and 1027 GO terms were significantly enriched at 10 and 15 DPI. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of the target genes of DEMs showed that only 5 and 3 pathways were significantly enriched at 10 and 15 DPI, respectively. The integrated analysis of miRNA–gene network revealed that the DEMs played important roles in the host immune and inflammatory responses to H. meleagridis infection by dynamically regulating the expression of immune and inflammation-related cytokines. Conclusion: Our results not only suggested that host miRNA expression was dynamically altered by H. meleagridis and host, but also revealed that more miRNAs and genes were involved in the later stage of the disease. In addition, host and H. meleagridis regulated the expression of immune and inflammation-related cytokines to respond to H. meleagridis infection. Our results will contribute to future research on miRNA-target interaction during H. meleagridis infection in chickens and provide new ideas for H. meleagridis control.
2022-01-21 | GSE193859 | GEO
Project description:Exoproteome from parasitic protists constitutes of extracellular proteins which play a fundamental role in multifactorial host-parasite interactions. Lytic factors, especially secreted proteases in extracellular milieu, are capable to modulate tissue invasion, thereby aggravating host susceptibility. Despite the important role of exoproteins during infection, exoproteomic data on Histomonas meleagridis are non-existent. The present study employed traditional 1D-in-gel-zymography (1D-IGZ) and micro-LC-ESI-MS/MS (shotgun proteomics), to scrutinize H. meleagridis exoproteomes, obtained from a clonal virulent and attenuated strains. Both strains were maintained as mono-eukaryotic monoxenic culture with Escherichia coli. We demonstrated active in vitro secretion kinetics of proteases by both parasites, with widespread proteolytic activity ranging from 17 kDa to 120 kDa. Based on protease inhibitor-susceptibility tests, a predominant repertoire of cysteine proteolysis was present in the parasite exoproteomes, with stronger activity from virulent H. meleagridis. Shotgun proteomics, aided by customized database, identified 176 proteins including actin, potential moonlighting glycolytic enzymes, lytic molecules such as pore-forming proteins (PFPs) and proteases like cathepsin-L like cysteine protease. To quantify the exoproteomic differences between the virulent and the attenuated H. meleagridis cultures, a sequential window acquisition of all theoretical spectra mass spectrometric (SWATH-MS) approach was applied. Surprisingly results showed most of the exoproteomic differences to be of bacterial origin, involving metabolism and locomotion. By deciphering such molecular signatures, novel insights into an inherent complex in vitro protozoan- bacteria relationship was elucidated.
2019-02-11 | PXD012037 | Pride
Project description:Histomonas meleagridis can cause histomonosis in poultry. Due to the prohibition of effective drugs, the prevention and treatment of the disease requires new strategies. Questions about its pathogenic mechanisms and virulence factors remain puzzling. To address some of these issues, comparative proteomic analysis of TMT was performed on a virulent strain and its attenuated strain of Chinese chicken-origin . A total of 3494 proteins were identified in the experiment, of which 745 proteins were differentially expressed (fold change ≥1.2 or ≤0.8 and p＜0.05 ), 192 were up-regulated in virulent strain, and 553 were up-regulated in attenuated strain. Gene ontology (GO) analysis revealed that the differentially expressed proteins were mainly involved in organic material metabolic processes, primary metabolic processes and cellular metabolic processes, exerting protein binding functions, and most were distributed in the cytoplasm and organelle. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that these differentially expressed proteins were mainly involved in pathway of Spliceosome, Cytosolic DNA-sensing pathway, Ribosome biogenesis in eukaryotes, RNA transport and Ribosome. In addition, some differentially expressed proteins were quantified by parallel reverse monitoring (PRM) assay to verify the results of TMT analysis. The above results provide some candidate protein-coding genes for further functional verification, which will help to understand the molecular mechanism of pathogenicity and attenuation of H. meleagridis more comprehensively.
2022-11-23 | PXD038344 |