iTRAQ-Based Global Phosphoproteomics Reveals Novel Molecular Differences Between Toxoplasma gondii Strains of Different Genotypes

Wang, Ze-Xiang; Zhou, Chun-Xue; Calderón-Mantilla, Guillermo; Petsalaki, Evangelia; He, Jun-Jun; Song, Huwei; Elsheikha, Hany M.; Zhu, Xing‐Quan

doi:10.3389/fcimb.2019.00307

Cited by 17 publications

(21 citation statements)

References 79 publications

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“…KEGG pathway analysis showed that the identified phosphoproteins were annotated into pathways with direct relevance to toxoplasmosis and metabolism, such as "Glycerophospholipid metabolism" and "Metabolic pathways" (Table S3, Figure S1), which is consistent with the reported phosphoproteins function in the phosphoproteomic analysis of different T. gondii strains (Wang et al, 2019). This result suggests that phosphoprotein-mediated pathway regarding toxoplasmosis and metabolism may be critical for the pathogenesis of T. gondii.…”

Section: Discussionsupporting

confidence: 82%

“…For example, Wang et al analyzed the phosphoproteome of three different strains of T. gondii tachyzoites (RH strain, PRU stain, and PYS strain), and identified several hundreds of phosphoproteins. This phosphoproteomic analysis is therefore beneficial for explaining the contribution of phosphorylation in regulating the virulence heterogeneity of the three parasite strains ( Wang et al, 2019 ). In our study, however, only 38 phosphoproteins were identified, which were much less than the reported number of phosphoproteins in T. gondii tachyzoites.…”

Section: Discussionmentioning

confidence: 99%

“…In our study, however, only 38 phosphoproteins were identified, which were much less than the reported number of phosphoproteins in T. gondii tachyzoites. The discrepancy between the number of identified T. gondii phosphoproteins may be due to the far below amount of loading protein of T. gondii for the analysis performed with the same approach, completely different lytic cycles of tachyzoites and various sources of parasite collection ( Wang et al, 2019 ). Since the phosphoproteome data generated in this study was confirmed by western blot from the aspect of human cells ( He et al, 2019 ), the phosphoproteome data reported in this study therefore is very reliable.…”

Section: Discussionmentioning

confidence: 99%

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iTRAQ-Based Phosphoproteomic Analysis of Toxoplasma gondii Tachyzoites Provides Insight Into the Role of Phosphorylation for its Invasion and Egress

Pan

et al. 2020

Front. Cell. Infect. Microbiol.

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The invasion and egress are two key steps in lytic cycle vital to the propagation of Toxoplasma gondii infection, and phosphorylation is believed to play important roles in these processes. However, the phosphoproteome of T. gondii at these two stages has not been characterized. In this study, we profiled the phosphoproteome of tachyzoites at the stages of “just invading” (JI) and “prior to egress” (PE) based on iTRAQ quantitative analysis, in which a total of 46 phosphopeptides, 42 phosphorylation sites, and 38 phosphoproteins were detected. In the comparison of PE vs. JI, 10 phosphoproteins were detected with their phosphorylation level significantly changed, and four of them were demonstrated to be significantly down-regulated at the transcriptional level. Bioinformatic analysis of these identified phosphoproteins suggested that phosphorylation-mediated modulation of protein function was employed to regulate the pathway of toxoplasmosis and metabolism and cellular processes correlated with tachyzoite’s binding, location, and metabolism, and thus play vital roles in the parasite lytic cycle. Moreover, cytoskeletal network (CN)-associated Inner Membrane Complex (IMC1, IMC4, IMC6 and IMC12), Intravascular Network (IVN)-related GRAs (GRA2, GRA3, GRA7 and GRA12), and Parasitophorous Vacuole Membrane (PVM)-localized ROP5 were shown to be enriched at the central nodes in the protein interaction network generated by bioinformatic analysis, in which the phosphorylation level of IMC4, GRA2, GRA3, and GRA12 were found to be significantly regulated. This study revealed the main cellular processes and key phosphoproteins crucial for the invasion and egress of T. gondii, which will provide new insights into the developmental biology of T. gondii in vitro and contribute to the understanding of pathogen-host interaction from the parasite perspective.

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Section: Discussionsupporting

confidence: 82%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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iTRAQ-Based Phosphoproteomic Analysis of Toxoplasma gondii Tachyzoites Provides Insight Into the Role of Phosphorylation for its Invasion and Egress

Pan

et al. 2020

Front. Cell. Infect. Microbiol.

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“…In this context, TgWNGI, a parasite specific kinase, has been shown to phosphorylate and regulate the association of GRAs such as GRA17 with the membrane (Beraki et al, 2019). However, GRA60 is not reported to be phosphorylated in any of the available phosphoproteome analyses (Z. X. Wang, Zhou, et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

Toxoplasma gondii GRA60 is an effector protein that modulates host cell autonomous immunity and contributes to virulence

Nyonda

Hammoudi

et al. 2020

Cellular Microbiology

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Toxoplasma gondii infects virtually any nucleated cell and resides inside a non‐phagocytic vacuole surrounded by a parasitophorous vacuolar membrane (PVM). Pivotal to the restriction of T. gondii dissemination upon infection in murine cells is the recruitment of immunity regulated GTPases (IRGs) and guanylate binding proteins (GBPs) to the PVM that leads to pathogen elimination. The virulent T. gondii type I RH strain secretes a handful of effectors including the dense granule protein GRA7, the serine–threonine kinases ROP17 and ROP18, and a pseudo‐kinase ROP5, that synergistically inhibit the recruitment of IRGs to the PVM. Here, we characterise GRA60, a novel dense granule effector, which localises to the vacuolar space and PVM and contributes to virulence of RH in mice, suggesting a role in the subversion of host cell defence mechanisms. Members of the host cell IRG defence system Irgb10 and Irga6 are recruited to the PVM of RH parasites lacking GRA60 as observed previously for the avirulent RHΔrop5 mutant, with RH preventing such recruitment. Deletion of GRA60 in RHΔrop5 leads to a recruitment of IRGs comparable to the single knockouts. GRA60 therefore represents a novel parasite effector conferring resistance to IRGs in type I parasites, and found associated to ROP18, a member of the virulence complex.

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“…The sensitivity of tissue culture and mouse inoculation methods for the demonstration of T. gondii organisms were found to be equal [24]. In addition, many studies prepared tachyzoites using mice inoculation [15,25,26,27,28,29]. The purpose of our research was to investigate acetylomic signatures of different genotypes of Toxoplasma gondii purified in vivo .…”

Section: Methodsmentioning

confidence: 99%

Label-Free Quantitative Acetylome Analysis Reveals Toxoplasma gondii Genotype-Specific Acetylomic Signatures

Wang

Zhou

et al. 2019

Microorganisms

Self Cite

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Distinct genotypic and pathogenic differences exist between Toxoplasma gondii genotypes. For example, genotype I is highly virulent, whereas genotype II and genotype III are less virulent. Moreover, Chinese 1 genotype (ToxoDB#9) is also virulent. Here, we compare the acetylomes of genotype 1 (RH strain) and Chinese 1 genotype (ToxoDB#9, PYS strain) of T. gondii. Using mass spectrometry enriched for acetylated peptides, we found a relationship between the levels of protein acetylation and parasite genotype-specific virulence. Notably, lysine acetylation was the largest (458 acetylated proteins) in RH strain, followed by PYS strain (188 acetylated proteins), whereas only 115 acetylated proteins were detected in PRU strain. Our analysis revealed four, three, and four motifs in RH strain, PRU strain and PYS strain, respectively. Three conserved sequences around acetylation sites, namely, xxxxxKAcHxxxx, xxxxxKAcFxxxx, and xxxxGKAcSxxxx, were detected in the acetylome of the three strains. However, xxxxxKAcNxxxx (asparagine) was found in RH and PYS strains but was absent in PRU strain. Our analysis also identified 15, 3, and 26 differentially expressed acetylated proteins in RH strain vs. PRU strain, PRU strain vs. PYS strain and PYS strain vs. RH strain, respectively. KEGG pathway analysis showed that a large proportion of the acetylated proteins are involved in metabolic processes. Pathways for the biosynthesis of secondary metabolites, biosynthesis of antibiotics and microbial metabolism in diverse environments were featured in the top five enriched pathways in all three strains. However, acetylated proteins from the virulent strains (RH and PYS) were more enriched in the pyruvate metabolism pathway compared to acetylated proteins from PRU strain. Increased levels of histone-acetyl-transferase and glycyl-tRNA synthase were detected in RH strain compared to PRU strain and PYS strain. Both enzymes play roles in stress tolerance and proliferation, key features in the parasite virulence. These findings reveal novel insight into the acetylomic profiles of major T. gondii genotypes and provide a new important resource for further investigations of the roles of the acetylated parasite proteins in the modulation of the host cell response to the infection of T. gondii.

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iTRAQ-Based Global Phosphoproteomics Reveals Novel Molecular Differences Between Toxoplasma gondii Strains of Different Genotypes

Cited by 17 publications

References 79 publications

iTRAQ-Based Phosphoproteomic Analysis of Toxoplasma gondii Tachyzoites Provides Insight Into the Role of Phosphorylation for its Invasion and Egress

iTRAQ-Based Phosphoproteomic Analysis of Toxoplasma gondii Tachyzoites Provides Insight Into the Role of Phosphorylation for its Invasion and Egress

Toxoplasma gondii GRA60 is an effector protein that modulates host cell autonomous immunity and contributes to virulence

Label-Free Quantitative Acetylome Analysis Reveals Toxoplasma gondii Genotype-Specific Acetylomic Signatures

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