Transcriptome sequencing of WT and chloroquine resistance transporter (TgCRT) ortholog-deficient Toxoplasma parasites
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PROVIDER: PRJNA478951 | ENA |
REPOSITORIES: ENA
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Project description:Purpose: TgCRT spans the membrane of the digestive vacuole of Toxoplasma parasites. The deletion of TgCRT causes the swelling of the digestive vacuole, which could interfere with the physiology and function of the VAC. The goals of this study are to compare transcriptome profiling (RNA-Seq) of WT and TgCRT-deletion strains to understand what strategies Toxoplasma takes to regulate the size and morphology of the digestive vacuole to help parasite disseminate infection. Methods: WT and TgCRT-deficient parasites were grown in human foreskin fibroblast cells for 2 days. The parasites were syringed, filter-purified, and resuspended in ice-cold PBS buffer. A trizol-based method was used to extract the total RNA from both strains. Total RNA was subjected to cDNA library construction using Illumina TruSeq RNA sample prep kit and Hi-Seq transcriptome sequencing. Results: Approximately 20 million sequence reads per sample were mapped to the genome of Toxoplasma GT1 strain (release 34). The differential gene analysis identified 102 genes with a fold-change ≥1.5 and FDR <0.05. Conclusions: Our study found that the ∆crt parasites reduce the transcript levels of several endolysosomal proteases, which could alleviate the proteolytic activities within the VAC to minimize the swelling. In addition, the RNA-Seq analysis also suggests that the parasites may activate their signaling pathways to manage the stress caused by the swelled VAC and increase other transporters to regulate the morphology of the digestive vacuole. Grant title: Regulation of proteolytic activity within a digestive vacuole in Toxoplasma gondii, the most common pathogen causing infectious posterior uveitis in infants and children Name of the funding source: Knights Templar Eye Foundation Principal Investigator: Dr. Zhicheng Dou
2019-04-29 | GSE116539 | GEO
Project description:Purpose: Toxoplasma must import inorganic phosphate via a transporter at the plasma membrane. Methods: WT and TgPiT-deficient parasites were grown in human foreskin fibroblasts for 2 days. The parasites were syringed, filter-purified, and resuspended n ice-cold PBS. A trizol-based method was used to extract total RNA from both strains. Toral RNA was subjected to cDNA library construction using Ilumina. Results: The parasite expresses a unique TgPiT as selective Pi transporter, located at the plasma membrane and on VAC internal membranes of endosomal organelles. Conclusion: TgPiT is essential for parasite survival, virulence, ionic homeostasis and osmoregulation
2021-03-01 | GSE145660 | GEO
Project description:Transcriptome sequencing of WT and inorganic phosphate transporter (TgPiT) ortholog-deficient Toxoplasma parasites
| PRJNA607846 | ENA
Project description:Apicomplexa are intracellular parasites that cause human and animal disease. They proliferate by a unique mechanism that combines closed mitosis with daughter cell budding. In past work we demonstrated that the Toxoplasma gondii centromeres are sequestered to the nuclear periphery proximal to the centrosome throughout the cell cycle. Here we show that interphase centromere clustering is not mediated by the mitotic spindle. Instead we propose a chromatin model of centromere clustering and define structural maintenance of chromosomes protein 1 (SMC1) as a persisting centromeric protein. We biochemically identify proteins that physically interact with SMC1, and CenH3 (a centromeric histone); prominent among those are predicted peripheral soluble components of the nuclear pore complex including TgExportin1. Treatment of parasites with the highly specific exportin1 inhibitor leptomycin B causes the dispersal of centromeres. Our results suggest that the nuclear envelope, and in particular peripheral components of the nuclear transport machinery orchestrate centromere positioning and parasite nuclear architecture. Chromatin immunoprecipitation (ChIP) of TgSMC1, three replicates applied to two microarrays each representing half of the Toxoplasma genome
2014-09-27 | E-GEOD-61806 | biostudies-arrayexpress
Project description:Apicomplexa are intracellular parasites that cause human and animal disease. They proliferate by a unique mechanism that combines closed mitosis with daughter cell budding. In past work we demonstrated that the Toxoplasma gondii centromeres are sequestered to the nuclear periphery proximal to the centrosome throughout the cell cycle. Here we show that interphase centromere clustering is not mediated by the mitotic spindle. Instead we propose a chromatin model of centromere clustering and define structural maintenance of chromosomes protein 1 (SMC1) as a persisting centromeric protein. We biochemically identify proteins that physically interact with SMC1, and CenH3 (a centromeric histone); prominent among those are predicted peripheral soluble components of the nuclear pore complex including TgExportin1. Treatment of parasites with the highly specific exportin1 inhibitor leptomycin B causes the dispersal of centromeres. Our results suggest that the nuclear envelope, and in particular peripheral components of the nuclear transport machinery orchestrate centromere positioning and parasite nuclear architecture.
2014-09-27 | GSE61806 | GEO
Project description:ITDR MS-CETSA was conducted on chloroquine-treated live parasites and lysate of P. falciparum trophozoites in order to identify potential drug protein targets.
2023-12-11 | PXD046662 | Pride
Project description:Infection of RAW264.7 cells with RH?ku80 parasites or mock-infection for 24 hours To measure changes in gene expression induced in macropahges upon Toxoplasma infection, we infected RAW 264.7 macrophages in cell culture with RH?ku80 parasites or syringe-lysed human foreskin fibroblast monolayers (mock-infected). RNA was harvested 24 hours post infection. Cells were infected with Toxoplasma or mock-infected in vitro, in duplicate
2014-02-25 | E-GEOD-55298 | biostudies-arrayexpress
Project description:Acetyl-CoA participates in post-translational modification of proteins, central carbon and lipid metabolism in several cell compartments. In mammals, the acetyl-CoA transporter 1 (AT1) facilitates the flux of cytosolic acetyl-CoA into the endoplasmic reticulum (ER), enabling the acetylation of proteins of the secretory pathway, in concert with dedicated acetyltransferases including Nat8. However, the implication of the ER acetyl-CoA pool in acetylation of ER-transiting proteins and their relevance throughout the parasites’ life cycle is unknown. Here, we evaluated the impact of blocking putative cytosolic acetyl-CoA import on acetylation of proteins through KO of the homologue of AT1 in the parasite Toxoplasma gondii.
2022-07-25 | PXD030932 | Pride
Project description:Infection of RAW264.7 cells with RHΔku80 parasites or mock-infection for 24 hours To measure changes in gene expression induced in macropahges upon Toxoplasma infection, we infected RAW 264.7 macrophages in cell culture with RHΔku80 parasites or syringe-lysed human foreskin fibroblast monolayers (mock-infected). RNA was harvested 24 hours post infection.
2014-02-25 | GSE55298 | GEO
Project description:Pyronaridine (PN) and chloroquine (CQ) are structurally related anti-malarial drugs with primarily the same mode of action. However, PN is effective against several multidrug-resistant lines of Plasmodium falciparum, including CQ-resistant lines, suggestive of important operational differences between the two drugs. Synchronized trophozoite-stage cultures of P. falciparum strain K1 (CQ resistant) were exposed to 50% inhibitory concentrations (IC50) of PN and CQ, and parasites were harvested from culture after 4 and 24 hours exposure. Global transcriptional changes effected by drug treatment were investigated using DNA microarrays.
2011-08-02 | GSE30867 | GEO