Comparative Phosphoproteomic Analysis of Sporulated Oocysts and Tachyzoites of <i>Toxoplasma gondii</i> Reveals Stage-Specific Patterns

<i>Toxoplasma gondii</i> is an obligate intracellular protozoan of severe threat to humans and livestock, whose life history harbors both gamic and apogamic stages. Chinese 1 (ToxoDB#9) was a preponderant genotype epidemic in food-derived animals and humans in China, with a different pat...

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Main Authors: Ze-Xiang Wang, Liang Che, Rui-Si Hu, Xiao-Lin Sun
Format: Article
Language:English
Published: MDPI AG 2022-02-01
Series:Molecules
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Online Access:https://www.mdpi.com/1420-3049/27/3/1022
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author Ze-Xiang Wang
Liang Che
Rui-Si Hu
Xiao-Lin Sun
author_facet Ze-Xiang Wang
Liang Che
Rui-Si Hu
Xiao-Lin Sun
author_sort Ze-Xiang Wang
collection DOAJ
description <i>Toxoplasma gondii</i> is an obligate intracellular protozoan of severe threat to humans and livestock, whose life history harbors both gamic and apogamic stages. Chinese 1 (ToxoDB#9) was a preponderant genotype epidemic in food-derived animals and humans in China, with a different pathogenesis from the strains from the other nations of the world. Posttranslational modifications (PTMs) of proteins were critical mediators of the biology, developmental transforms, and pathogenesis of protozoan parasites. The phosphoprotein profiling and the difference between the developmental phases of <i>T. gondii</i>, contributing to development and infectivity, remain unknown. A quantitative phosphoproteomic approach using IBT integrated with TiO<sub>2</sub> affinity chromatography was applied to identify and analyze the difference in the phosphoproteomes between the sporulated oocysts and the tachyzoites of the virulent ToxoDB#9 (PYS) strain of <i>T. gondii</i>. A total of 4058 differential phosphopeptides, consisting of 2597 upregulated and 1461 downregulated phosphopeptides, were characterized between sporulated the oocysts and tachyzoites. Twenty-one motifs extracted from the upregulated phosphopeptides contained 19 serine motifs and 2 threonine motifs (GxxTP and TP), whereas 16 motifs identified from downregulated phosphopeptides included 13 serine motifs and 3 threonine motifs (KxxT, RxxT, and TP). Beyond the traditional kinases, some infrequent classes of kinases, including Ab1, EGFR, INSR, Jak, Src and Syk, were found to be corresponding to motifs from the upregulated and downregulated phosphopeptides. Remarkable functional properties of the differentially expressed phosphoproteins were discovered by GO analysis, KEGG pathway analysis, and STRING analysis. S8GFS8 (DNMT1-RFD domain-containing protein) and S8F5G5 (Histone kinase SNF1) were the two most connected peptides in the kinase-associated network. Out of these, phosphorylated modifications in histone kinase SNF1 have functioned in mitosis and interphase of <i>T. gondii</i>, as well as in the regulation of gene expression relevant to differentiation. Our study discovered a remarkable difference in the abundance of phosphopeptides between the sporulated oocysts and tachyzoites of the virulent ToxoDB#9 (PYS) strain of <i>T. gondii</i>, which may provide a new resource for understanding stage-specific differences in PTMs and may enhance the illustration of the regulatory mechanisms contributing to the development and infectivity of <i>T. gondii</i>.
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spelling doaj.art-219d5703b14d43159842ea4be96598af2023-11-23T17:16:22ZengMDPI AGMolecules1420-30492022-02-01273102210.3390/molecules27031022Comparative Phosphoproteomic Analysis of Sporulated Oocysts and Tachyzoites of <i>Toxoplasma gondii</i> Reveals Stage-Specific PatternsZe-Xiang Wang0Liang Che1Rui-Si Hu2Xiao-Lin Sun3College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, ChinaLanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, ChinaCollege of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China<i>Toxoplasma gondii</i> is an obligate intracellular protozoan of severe threat to humans and livestock, whose life history harbors both gamic and apogamic stages. Chinese 1 (ToxoDB#9) was a preponderant genotype epidemic in food-derived animals and humans in China, with a different pathogenesis from the strains from the other nations of the world. Posttranslational modifications (PTMs) of proteins were critical mediators of the biology, developmental transforms, and pathogenesis of protozoan parasites. The phosphoprotein profiling and the difference between the developmental phases of <i>T. gondii</i>, contributing to development and infectivity, remain unknown. A quantitative phosphoproteomic approach using IBT integrated with TiO<sub>2</sub> affinity chromatography was applied to identify and analyze the difference in the phosphoproteomes between the sporulated oocysts and the tachyzoites of the virulent ToxoDB#9 (PYS) strain of <i>T. gondii</i>. A total of 4058 differential phosphopeptides, consisting of 2597 upregulated and 1461 downregulated phosphopeptides, were characterized between sporulated the oocysts and tachyzoites. Twenty-one motifs extracted from the upregulated phosphopeptides contained 19 serine motifs and 2 threonine motifs (GxxTP and TP), whereas 16 motifs identified from downregulated phosphopeptides included 13 serine motifs and 3 threonine motifs (KxxT, RxxT, and TP). Beyond the traditional kinases, some infrequent classes of kinases, including Ab1, EGFR, INSR, Jak, Src and Syk, were found to be corresponding to motifs from the upregulated and downregulated phosphopeptides. Remarkable functional properties of the differentially expressed phosphoproteins were discovered by GO analysis, KEGG pathway analysis, and STRING analysis. S8GFS8 (DNMT1-RFD domain-containing protein) and S8F5G5 (Histone kinase SNF1) were the two most connected peptides in the kinase-associated network. Out of these, phosphorylated modifications in histone kinase SNF1 have functioned in mitosis and interphase of <i>T. gondii</i>, as well as in the regulation of gene expression relevant to differentiation. Our study discovered a remarkable difference in the abundance of phosphopeptides between the sporulated oocysts and tachyzoites of the virulent ToxoDB#9 (PYS) strain of <i>T. gondii</i>, which may provide a new resource for understanding stage-specific differences in PTMs and may enhance the illustration of the regulatory mechanisms contributing to the development and infectivity of <i>T. gondii</i>.https://www.mdpi.com/1420-3049/27/3/1022<i>Toxoplasma gondii</i>life cycleIBTphosphoproteomics
spellingShingle Ze-Xiang Wang
Liang Che
Rui-Si Hu
Xiao-Lin Sun
Comparative Phosphoproteomic Analysis of Sporulated Oocysts and Tachyzoites of <i>Toxoplasma gondii</i> Reveals Stage-Specific Patterns
Molecules
<i>Toxoplasma gondii</i>
life cycle
IBT
phosphoproteomics
title Comparative Phosphoproteomic Analysis of Sporulated Oocysts and Tachyzoites of <i>Toxoplasma gondii</i> Reveals Stage-Specific Patterns
title_full Comparative Phosphoproteomic Analysis of Sporulated Oocysts and Tachyzoites of <i>Toxoplasma gondii</i> Reveals Stage-Specific Patterns
title_fullStr Comparative Phosphoproteomic Analysis of Sporulated Oocysts and Tachyzoites of <i>Toxoplasma gondii</i> Reveals Stage-Specific Patterns
title_full_unstemmed Comparative Phosphoproteomic Analysis of Sporulated Oocysts and Tachyzoites of <i>Toxoplasma gondii</i> Reveals Stage-Specific Patterns
title_short Comparative Phosphoproteomic Analysis of Sporulated Oocysts and Tachyzoites of <i>Toxoplasma gondii</i> Reveals Stage-Specific Patterns
title_sort comparative phosphoproteomic analysis of sporulated oocysts and tachyzoites of i toxoplasma gondii i reveals stage specific patterns
topic <i>Toxoplasma gondii</i>
life cycle
IBT
phosphoproteomics
url https://www.mdpi.com/1420-3049/27/3/1022
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AT liangche comparativephosphoproteomicanalysisofsporulatedoocystsandtachyzoitesofitoxoplasmagondiiirevealsstagespecificpatterns
AT ruisihu comparativephosphoproteomicanalysisofsporulatedoocystsandtachyzoitesofitoxoplasmagondiiirevealsstagespecificpatterns
AT xiaolinsun comparativephosphoproteomicanalysisofsporulatedoocystsandtachyzoitesofitoxoplasmagondiiirevealsstagespecificpatterns