Transcriptome sequencing of WT and inorganic phosphate transporter (TgPiT) ortholog-deficient Toxoplasma parasites
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ABSTRACT:
PROVIDER: PRJNA607846 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR11127604_1.fastq.gz | Fastqsanger.gz | |||
| SRR11127604_2.fastq.gz | Fastqsanger.gz | |||
| SRR11127605_1.fastq.gz | Fastqsanger.gz | |||
| SRR11127605_2.fastq.gz | Fastqsanger.gz | |||
| SRR11127606_1.fastq.gz | Fastqsanger.gz |
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Similar Datasets
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 chloroquine resistance transporter (TgCRT) ortholog-deficient Toxoplasma parasites
| PRJNA478951 | ENA
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:Toxoplasma gondii is an obligate intracellular parasite that acquires all necessary nutrients from the hosts, but the exact nutrient acquisition mechanisms are poorly understood. Here, we identified three putative phosphate transporters in T. gondii. TgPiT and TgPT2 are mainly on the plasma membrane, whereas TgmPT is localized to the mitochondrion. TgPiT and TgmPT are widely present and conserved in apicomplexan parasites that include Plasmodium and Eimeria species. Nonetheless, they are dispensable for the growth and virulence of Toxoplasma. TgPT2, on the other hand, is restricted to coccidia parasites and is essential for Toxoplasma survival. TgPT2 depletion led to reduced motility and invasion, as well as growth arrest of the parasites both in vitro and in vivo. Both TgPiT and TgPT2 have phosphate transport activities and contribute to parasites' inorganic phosphate (Pi) absorption. Interestingly, the Pi importing activity of Toxoplasma parasites could be competitively inhibited by ATP and AMP. Furthermore, direct uptake of 32P-ATP was also observed, indicating the parasites' ability to scavenge host ATP. Nonetheless, ATP/AMP import is not mediated by TgPiT or TgPT2, suggesting additional mechanisms. Together, these results show the complex pathways of phosphate transport in Toxoplasma, and TgPT2 is a potential target for antitoxoplasmic intervention design due to its essential role in parasite growth. IMPORTANCE To grow and survive within host cells, Toxoplasma must scavenge necessary nutrients from hosts to support its parasitism. Transporters located in the plasma membrane of the parasites play critical roles in nutrient acquisition. Toxoplasma encodes a large number of transporters, but so far, only a few have been characterized. In this study, we identified two phosphate transporters, TgPiT and TgPT2, to localize to the plasma membrane of Toxoplasma. Although both TgPiT and TgPT2 possess phosphate transport activities, only the novel transporter TgPT2 was essential for parasite growth, both in vitro and in vivo. In addition, TgPT2 and its orthologs are only present in coccidia parasites. As such, TgPT2 represents a potential target for drug design against toxoplasmosis. In addition, our data indicated that Toxoplasma can take up ATP and AMP from the environment, providing new insights into the energy metabolism of Toxoplasma.
| S-EPMC9604053 | biostudies-literature
Project description:One of the main regulators of phosphate homeostasis is fibroblast growth factor 23 (FGF23), secreted by osteocytes. The effects of organic versus inorganic dietary phosphate on this homeostasis is unclear. This study used MC3T3-E1 osteocyte-like cells to examine the transcriptomic responses to these phosphates. Most importantly, the expression and secretion of FGF23 was only increased in response to organic phosphate. Gene ontology terms related to a response to environmental change were only enriched in osteocytes treated with organic phosphate while osteocytes treated with inorganic phosphate were enriched for terms associated with regulation of cellular phosphate metabolism. Inhibition of MAPK signaling diminished the response of Fgf23 to organic phosphate, suggesting it activates FGF23. TGF-β signaling inhibition increased Fgf23 expression after the addition of organic phosphate, while the negative TGF-β regulator Skil decreased this response. In summary, the observed differential response of osteocytes to phosphate types may have consequences for phosphate homeostasis.
2024-03-18 | GSE245758 | GEO
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: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:Acclimation of Nodularia spumigena CCY9914 to inorganic phosphate limitation
| PRJNA880843 | ENA
Project description:The liver plays a central role in whole-body lipid and glucose homeostasis. Increasing dietary fat intake results in increased hepatic fat deposition, which is associated with a risk for development of insulin resistance and type 2 diabetes. In this study, we demonstrate a role for the phosphate inorganic transporter 1 (PiT1/SLC20A1) in regulating metabolism. Specific knockout of Pit1 in hepatocytes significantly improved glucose tolerance and insulin sensitivity, enhanced insulin signalling, and decreased hepatic lipogenesis. We identified USP7 as a PiT1 binding partner and demonstrated that Pit1 deletion inhibited USP7/IRS1 dissociation upon insulin stimulation. This prevented IRS1 ubiquitination and its subsequent proteasomal degradation. As a consequence delayed insulin negative feedback loop and sustained insulin signalling were observed. Moreover, PiT1-deficient mice were protected against high fat diet-induced obesity and diabetes. Our findings indicate that PiT1 has potential as a therapeutic target in the context of metabolic syndrome, obesity, and diabetes.
2016-08-25 | E-MTAB-4989 | biostudies-arrayexpress
Project description:In E. coli the phosphate homeostasis is regulated by the Pst system and the two-component system PhoB/R. Pathogens like E. coli O157:H7 are responsible for many outbreaks and can be found and survive in poor inorganic phosphate (Pi) environments. To understand global EHEC O157:H7 EDL933 strain responses to Pi-starvation, we compared the transcriptomes of EDL933 the WT strain grown in MOPS Pi rich medium and that grown in MOPS Pi poor medium, using the Affymetrix GeneChip® E. coli Genome 2.0 Array. Also we investigated the EDL933 global response to the absence of PhoB by comparing the transcriptomes of the WT strain the ΔphoB mutant both grown in Low Pi
2014-03-15 | GSE50529 | GEO