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Transcriptome sequencing of WT and inorganic phosphate transporter (TgPiT) ortholog-deficient Toxoplasma parasites

ABSTRACT: 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

ORGANISM(S): Toxoplasma gondii

PROVIDER: GSE145660 | GEO | 2021/03/01

REPOSITORIES: GEO

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Transcriptome sequencing of WT and chloroquine resistance transporter (TgCRT) ortholog-deficient Toxoplasma parasites

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

The Plasmodium falciparum protein PfMRP1 functions as an influx ABC transporter

Project description:Adenosine triphosphate (ATP)-binding cassette (ABC) transporters play an important role in mediating solute or drug transport across cellular membranes. Although this class of transporters has been well characterized in diverse organisms little is known about the physiological roles in Plasmodium falciparum, the deadliest malaria parasite species. We studied the Plasmodium falciparum Multidrug Resistance-associated Protein 1 (PfMRP1; PF3D7_0112200), an ABC transporter localized to the parasite plasma membrane, generating genetic disrupted parasites. We demonstrate that parasites with disrupted pfmrp1 are resistant to folate analogs, methotrexate and aminopterin, with antimalarial activity. This phenotype occurs due to reduction in compound accumulation in the parasite cytoplasm. Phylogenetic analysis supports pfmrp1 being distantly related to ABC transporters in other eukaryotes, suggesting an unusual function. We propose that PfMRP1 can act as a solute importer, a function not previously observed in this organism.

2022-01-31 | GSE195649 | GEO

Transcriptome sequencing of WT and inorganic phosphate transporter (TgPiT) ortholog-deficient Toxoplasma parasites

Project description:Transcriptome sequencing of WT and inorganic phosphate transporter (TgPiT) ortholog-deficient Toxoplasma parasites

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Malaria parasite manipulates erythrocyte membranes for intracellular adaptation : RNA-seq

Project description:The ability to respond to stressful host environments is crucial for the survival of intracellular pathogens, during which transporters play key roles in the nutrient scavenging, such as lipid uptake. Here we discover a conserved malarial gene that orchestrates the parasitic adaption in the terminally differentiated, phospholipid scarce erythrocytes. The expression level of this gene, here named CAP, is the intensively upregulated in the erythrocyte-infecting parasites than the reticulocyte-infecting counterparts. We show that CAP encodes a small protein exported onto the membrane of infected erythrocytes where it interacts with host CTL1, a choline and ethanolamine transporter, and upregulates biosynthesis of phosphatidylcholine and phosphatidylethanolamine, the major component of parasite membrane. Deletion of CAP causes Plasmodium to upturn reticulocyte invasion and sexual stage differentiation. This work presents a novel mechanism for intracellular parasite to import lipid precursors into parasitized cells by manipulating an endogenous host transporter.

2024-01-12 | GSE252704 | GEO

A domino effect in drug action: From metabolic assault towards parasite differentiation

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2010-10-18 | E-GEOD-24275 | biostudies-arrayexpress

A domino effect in drug action: From metabolic assault towards parasite differentiation

2010-10-18 | GSE24275 | GEO

A G-protein-coupled receptor modulates gametogenesis via PKG-mediated signaling cascade in Plasmodium berghei

Project description:Gametogenesis is essential for malaria parasite transmission, but the molecular mechanism of this process remains to be refined. Here, we identified a G-protein-coupled receptor 180 (GPR180) that plays a critical role in signal transduction during gametogenesis in Plasmodium. P. berghei GPR180 was predominantly expressed in gametocytes and ookinetes and associated with the plasma membrane in female gametes and ookinetes. Knockout of pbgpr180 (Δpbgpr180) had no noticeable effect on blood-stage development but impaired gamete formation and reduced transmission of the parasites to mosquitoes. Transcriptome analysis revealed that a large proportion of the dysregulated genes in the Δpbgpr180 gametocytes had assigned functions in cyclic nucleotide signaling transduction. In the Δpbgpr180 gametocytes, the intracellular cGMP level was significantly reduced, and the cytosolic Ca2+ mobilization showed a delay and a reduction in the magnitude during gametocyte activation. These results suggest that PbGPR180 functions upstream of the cGMP-protein kinase G-Ca2+ signaling pathway. In line with this functional prediction, the PbGPR180 protein was found to interact with several transmembrane transporter proteins and the small GTPase Rab6 in activated gametocytes. Allele replacement of pbgpr180 with the P. vivax ortholog pvgpr180 showed equal competence of the transgenic parasite in sexual development, suggesting functional conservation of this gene in Plasmodium spp. Furthermore, an anti-PbGPR180 monoclonal antibody and the anti-PvGPR180 serum possessed robust transmission-blocking activities. These results indicate that GPR180 is involved in signal transduction during gametogenesis in malaria parasites and is a promising target for blocking parasite transmission.

2022-04-06 | GSE198287 | GEO

Transcriptome sequencing of WT and chloroquine resistance transporter (TgCRT) ortholog-deficient Toxoplasma parasites

Project description:Transcriptome sequencing of WT and chloroquine resistance transporter (TgCRT) ortholog-deficient Toxoplasma parasites

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Comparison of gene expression between wild type and AP2-L KO parasites

Project description:In the rodent malaria parasite Plasmodium berghei, we found that an AP2 family transcription factor designated AP2-L plays a critical role in the development of the liver stage. Using DNA microarray analysis we showed that the expression of several genes, including those of parasitophorous vacuole membrane proteins, was significantly decreased in the early liver stage of AP2-L-depleted parasites. Gene expression of P. berghei liver stages was compared between wild type and AP2-L KO parasites. Liver stage parasites were cultured for 24h using HepG2 cells. Ten and seven biologically independent experiments were performed for wild type and AP2-L KO parasites, respectively.

2012-04-30 | E-GEOD-34877 | biostudies-arrayexpress

sut2_vs_WT

Project description:Series of 12 repetitions of hybridization of treatment (sut2) and control (WT) each. Comparison of the Arabidopsis sut2 mutant lacking plasma-membrane sucrose sensor/transporter versus WT. L. Barker et al., Plant Cell 12 (2000), pp. 1153-64 Keywords: repeat sample

2010-06-10 | E-GEOD-1252 | biostudies-arrayexpress

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