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Toxoplasma gondii eIF-5A knockdown iTraq

ABSTRACT: Proliferative stage (tachyzoite) is a critical factor in the transmission and pathogenesis of Toxoplasma gondii (T. gondii), which is capable of infecting a large variety of host cells. Proto-oncogene eukaryotic translation initiation factor (eIF-5A) is thought to function in regulation of many cellular processes including cell proliferation. Investigation the specific roles of eIF-5A involved in T. gondii is instrumental for the elucidation of tachyzoite replication. Here, eIF-5A knockdown strain was generated and analyzed by isobaric tags for relative and absolute quantification (iTraq), which suggested that proto-oncogene eIF-5A also acts as an essential gene to ensure proper protein synthesis during T. gondii invasion and replication.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Toxoplasma Gondii Rh

SUBMITTER: xiangrui Li

LAB HEAD: Li xiangrui

PROVIDER: PXD016810 | Pride | 2021-09-09

REPOSITORIES: Pride

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			Action	DRS
	F18FTSECKF0107_rep1.dat	Other
	P180032_1.raw	Raw
	P180032_10.raw	Raw
	P180032_11.raw	Raw
	P180032_12.raw	Raw

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Publications

Proteomics analysis reveals that the proto-oncogene eIF-5A indirectly influences the growth, invasion and replication of Toxoplasma gondii tachyzoite.

Liu Xinchao X Li Chunjing C Li Xiaoyu X Ehsan Muhammad M Lu Mingmin M Li Ke K Xu Lixin L Yan Ruofeng R Song Xiaokai X Li XiangRui X

Parasites & vectors 20210526 1

<h4>Background</h4>The proliferative stage (tachyzoite) of Toxoplasma gondii (T. gondii) is critical for its transmission and pathogenesis, and a proto-oncogene eukaryotic translation initiation factor (eIF-5A) plays an important role in various cellular processes such as cell multiplication.<h4>Methods</h4>We performed a proteomic study to evaluate the specific roles of eIF-5A involved in invasion and replication of T. gondii, and both in vivo and in vitro trials using eIF-5A-interfered and wil ...[more]

PMID: 34039408

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2008-05-22 | E-GEOD-11515 | biostudies-arrayexpress

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Coordinated Progression Through Two Subtranscriptomes Underlies the Tachyzoite Cycle of Toxoplasma gondii

Project description:We describe the cell cycle transcriptome of the Toxoplasma gondii that has emerged as a major genetic model for the study of Apicomplexa parasites. Two distinct transcriptional waves accompany the relatively simple binary replication of tachyzoite stage (endodyogeny) functionally separating conserved gene expression in the eukaryotic G1 phase from the lineage-specific expression that predominates in the parasite S phase and mitotic periods. This division of transcriptional focus closely mirrors the intimate relationship that has evolved between mitosis and building of the daughter parasites and invasion organelles. Promoter mechanisms appear to orchestrate the induction of gene expression in dividing tachyzoites with up to two dozen cell cycle AP2 factors likely acting within a transcriptional regulatory network to coordinate the parasite cell cycle transcriptome. To characterize the cell cycle transcriptome of Toxoplasma tachyzoites, we expanded a thymidine-synchrony model to isolate sufficient RNA for microarray expression studies. The ToxoGeneChip microarray (http://ancillary.toxodb.org/docs/Array-Tutorial.html) was used to measure mRNA expression in 13 duplicate samples (R0 to R12) covering 12 hours post-synchronization and nearly two tachyzoite replication cycles.

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Project description:Toxoplasma gondii is an apicomplexan parasite infecting human and animals, causing huge health concerns and economic losses. However, it is unclear about the exact mechanism of T.gondii tachyzoite infected macrophage and macrophage resisted T.gondii, especially for local isolates such as TgHB1 isolated in China. Our study focused on the transcriptional difference of pig alveolar macrophages (3D4/21) infected with china isolated TgHB1 compared to TgRH and TgME49 toxoplasma gondii standard strains.

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Toxoplasma gondii MOB1 interactome

Project description:MOB1 is a conserved protein that regulates cellular proliferation versus apoptosis, centrosome duplication and cellular differentiation in multicellular eukaryotes and also cytokinesis and division axis orientation in unicellular and multicellular eukaryotes. Toxoplasma gondii, an obligate intracellular parasite of veterinary and medical importance, presents one MOB1 protein. T. gondii interconverts between several cellular stages during its life cycle, namely between fast replicating tachyzoite and slow replicating bradyzoite stages during its asexual cycle, a key ability for its success as a parasite. Bradyzoites produce tissue cysts, establishing a chronic infection that enables recrudescence. Conversion is dependent on cell cycle regulation and involves cell differentiation and regulation of replication. This led us to select MOB1 as a strong candidate to be involved in the Toxoplasma replication process. To elucidate how MOB1 acts in T. gondii, we employed a proximity biotinylation method and identified the MOB1 interactome. Toxoplasma gondii RH tachyzoites were transfected with BirA containing plasmid vectors for random integration and two strains were isolated. The control strain expresses a FLAG-BirA recombinant protein while the test strain expresses a FLAG-BirA-MOB1 recombinant protein. Biotinylated proteins were purified using streptavidin-agarose beads. The purified proteins were trypsinized and analyzed by nanoLC-MS/MS.

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The Toxoplasma protein phosphatase 6 catalytic subunit (TgPP6C) is essential for cell cycle progression and virulence

Project description:Protein phosphatases are post-translational regulators of Toxoplasma gondii proliferation, tachyzoite-bradyzoite differentiation and pathogenesis. Here, we identify the putative protein phosphatase 6 (TgPP6) subunits of T. gondii and elucidate their role in the parasite lytic cycle. The putative catalytic subunit TgPP6C and regulatory subunit TgPP6R likely form a complex whereas the predicted structural subunit TgPP6S, with low homology to the human PP6 structural subunit, does not coassemble with TgPP6C and TgPP6R. Functional studies showed that TgPP6C and TgPP6R are essential for parasite growth and replication. The ablation of TgPP6C significantly reduced the synchronous division of the parasite's daughter cells during endodyogeny, resulting in disordered rosettes. Moreover, the six conserved motifs of TgPP6C were required for efficient endodyogeny. Phosphoproteomic analysis revealed that ablation of TgPP6C predominately altered the phosphorylation status of proteins involved in the regulation of the parasite cell cycle. Deletion of TgPP6C significantly attenuated the parasite virulence in mice. Immunization of mice with TgPP6C-deficient type I RH strain induced protective immunity against challenge with a lethal dose of RH or PYS tachyzoites and Pru cysts. Taken together, the results show that TgPP6C contributes to the cell division, replication and pathogenicity in T. gondii.

2023-11-28 | PXD041907 | Pride

72hr Toxoplasma gondii bradyzoite induction

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Single-cell RNA sequencing (scRNA-seq) on Treg cells from PBS and STAg treated mice

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Transcription profiling of Toxoplasma gondii tachyzoite to bradyzoite differentiation mutants

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