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Profound effect of asparagine-linked glycosylation protein 11 (TgALG11) on genomic expression of Toxoplasma gondii

ABSTRACT: Toxoplasma gondii is an intracellular parasite that relies on complex molecular mechanisms to support its proliferation and survival. In this study, we investigate the role of TgALG11 in these processes and explore the underlying regulatory network. Our data demonstrate that TgALG11 is essential for maintaining T. gondii proliferation and for normal N-glycosylation. To further elucidate the molecular mechanisms underlying the essential functions of TgALG11, we performed transcriptome sequencing to identify the genes regulated by this factor. Total RNA was extracted from iTgALG11 strains cultured with 0.5 μg/ml ATc (knockdown strain) and DMSO (control), and subjected to deep sequencing using the Illumina Novaseq™ 6000 platform. The sequence reads were subsequently mapped to the T. gondii genome, resulting in a comprehensive transcriptome encompassing 8,948 genes. Among these, 934 genes were differentially expressed due to TgALG11 deficiency, including 315 downregulated and 5 upregulated genes (DESeq, |log2(FoldChange)| ≥ 1, padj ≤ 0.05). Functional enrichment analysis revealed that these genes were associated with various Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Our findings provide novel insights into the profound impact of TgALG11 on the T. gondii transcriptome, underscoring its critical role in parasite biology.

ORGANISM(S): Toxoplasma gondii

PROVIDER: GSE303275 | GEO | 2025/07/27

REPOSITORIES: GEO

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Project description:Toxoplasma gondii is a globally distributed obligate intracellular parasite which can cause zoonotic toxoplasmosis with great harms. The average death time of mice that infected with Toxoplasma gondii RH strain tachyzoites recovered from the liquid nitrogen was shortened after multiple generations. It has been reported that the parasite is in a state of static virulence during cryopreservation and the virulence of the protozoan parasite can be enhanced after continuous passages in hosts under laboratory conditions. However, no research has been conducted to elucidate its biological mechanism. Herein, we sequenced the T. gondii transcriptome using RNA-Seq technology and performed de novo assembly to investigated the virulence factors expression changes by comparing gene expression profiles between incipiently recovered and completely resuscitated tachyzoites. Transcriptome analysis identified 1,951 differentially expressed transcripts in infected liver, of which 1,752 were significantly downregulated and 199 upregulated. We identified many differentially expressed proteins and genes, including serine/threonine kinase, calnexin, myosin and microtubule-associated protein which have previously been reported to be either involved in cell adhesion, parasite gliding or participate in cell invasion. The great majority of the virulence factors including microneme proteins, rhoptry proteins and dense granule proteins were upregulated in fully recovered tachyzoites. The enhanced virulence of recovered Toxoplasma gondii RH strain from the liquid nitrogen is associated with the up-regulated expression of MICs, ROPs and GRAs. Our data will facilitate future genomic research and in-depth annotation of Toxoplasma gondii RH strain genomes. This study provides a profile of the candidate genes that are suspected to be involved with virulence enhancement of recovered Toxoplasma gondii RH strain tachyzoites. Many further studies should be carried out to confirm the function of the candidate genes. Moreover, the preliminary identification of genes and pathways exhibiting differential expression in complete resuscitation stage may further our general understanding of virulence enhancement in this parasite.

Project description:Tymoshenko2015 - Genome scale metabolic model - ToxoNet1 This model is described in the article: Metabolic Needs and Capabilities of Toxoplasma gondii through Combined Computational and Experimental Analysis. Tymoshenko S, Oppenheim RD, Agren R, Nielsen J, Soldati-Favre D, Hatzimanikatis V. PLoS Comput. Biol. 2015 May; 11(5): e1004261 Abstract: Toxoplasma gondii is a human pathogen prevalent worldwide that poses a challenging and unmet need for novel treatment of toxoplasmosis. Using a semi-automated reconstruction algorithm, we reconstructed a genome-scale metabolic model, ToxoNet1. The reconstruction process and flux-balance analysis of the model offer a systematic overview of the metabolic capabilities of this parasite. Using ToxoNet1 we have identified significant gaps in the current knowledge of Toxoplasma metabolic pathways and have clarified its minimal nutritional requirements for replication. By probing the model via metabolic tasks, we have further defined sets of alternative precursors necessary for parasite growth. Within a human host cell environment, ToxoNet1 predicts a minimal set of 53 enzyme-coding genes and 76 reactions to be essential for parasite replication. Double-gene-essentiality analysis identified 20 pairs of genes for which simultaneous deletion is deleterious. To validate several predictions of ToxoNet1 we have performed experimental analyses of cytosolic acetyl-CoA biosynthesis. ATP-citrate lyase and acetyl-CoA synthase were localised and their corresponding genes disrupted, establishing that each of these enzymes is dispensable for the growth of T. gondii, however together they make a synthetic lethal pair. This model is hosted on BioModels Database and identified by: MODEL1504280000. To cite BioModels Database, please use: BioModels Database: An enhanced, curated and annotated resource for published quantitative kinetic models. To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer to CC0 Public Domain Dedication for more information.

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Profound effect of asparagine-linked glycosylation protein 11 (TgALG11) on genomic expression of Toxoplasma gondii

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