Volume 97, Issue 1
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



The invasion of tachyzoites into the host cell results in extensive host cell signaling activation/deactivation that is usually regulated by the phosphorylation/dephosphorylation. To elucidate how regulates host cell signal transduction, the comparative phosphoproteome of stable isotope labeling with amino acids in cell culture–labeled human foreskin fibroblast cells was analyzed. The cells were grouped (Light [L], Medium [M], and Heavy [H] groups) based on the labeling isotope weight and were infected with for different lengths of time (L: 0 hour; M: 2 hours; and H: 6 hours). A total of 892 phosphoproteins were identified with 1,872 phosphopeptides and 1,619 phosphorylation sites. The M versus L comparison revealed 694 significantly regulated phosphopeptides (436 upregulated and 258 downregulated). The H versus L comparison revealed 592 significantly regulated phosphopeptides (146 upregulated and 446 downregulated). The H versus M comparison revealed 794 significantly regulated phosphopeptides (149 upregulated and 645 downregulated). At 2 and 6 hours post- infection, the most predominant host cell reactions were cell cycle regulation and cytoskeletal reorganization, which might be required for the efficient invasion and multiplication of . Similar biological process profiles but different molecular function categories of host cells infected with for 2 and 6 hours, which suggested that the host cell processes were not affected significantly by infection but emphasized some differences in specific cellular processes at this two time points. Western blotting verification of some significantly regulated phosphoprotein phosphorylation sites was consistent with the mass spectra data. This study provided new insights into and further understanding of pathogen–host interactions from the host cell perspective.


Article metrics loading...

Loading full text...

Full text loading...



  1. Hotez PJ, , 2014. Neglected parasitic infections and poverty in the United States. PLoS Negl Trop Dis 8: e3012.[Crossref]
  2. Laliberte J, Carruthers VB, , 2008. Host cell manipulation by the human pathogen Toxoplasma gondii . Cell Mol Life Sci 65: 19001915.[Crossref]
  3. Weiss LM, Dubey JP, , 2009. Toxoplasmosis: a history of clinical observations. Int J Parasitol 39: 895901.[Crossref]
  4. Peng HJ, Chen XG, Lindsay DS, , 2011. A review: competence, compromise, and concomitance-reaction of the host cell to Toxoplasma gondii infection and development. J Parasitol 97: 620628.[Crossref]
  5. Olsen JV, Blagoev B, Gnad F, Macek B, Kumar C, Mortensen P, Mann M, , 2006. Global, in vivo, and site-specific phosphorylation dynamics in signaling networks. Cell 127: 635648.[Crossref]
  6. Cohen P, , 2001. The role of protein phosphorylation in human health and disease. The Sir Hans Krebs Medal Lecture. Eur J Biochem 268: 50015010.[Crossref]
  7. Song KJ, Ahn HJ, Nam HW, , 2012. Anti-apoptotic effects of SERPIN B3 and B4 via STAT6 activation in macrophages after infection with Toxoplasma gondii . Korean J Parasitol 50: 16.[Crossref]
  8. Peixoto L, Chen F, Harb OS, Davis PH, Beiting DP, Brownback CS, Ouloguem D, Roos DS, , 2010. Integrative genomic approaches highlight a family of parasite-specific kinases that regulate host responses. Cell Host Microbe 8: 208218.[Crossref]
  9. Vutova P, Wirth M, Hippe D, Gross U, Schulze-Osthoff K, Schmitz I, Luder CG, , 2007. Toxoplasma gondii inhibits Fas/CD95-triggered cell death by inducing aberrant processing and degradation of caspase 8. Cell Microbiol 9: 15561570.[Crossref]
  10. Melo MB, Nguyen QP, Cordeiro C, Hassan MA, Yang N, McKell R, Rosowski EE, Julien L, Butty V, Darde ML, Ajzenberg D, Fitzgerald K, Young LH, Saeij JP, , 2013. Transcriptional analysis of murine macrophages infected with different Toxoplasma strains identifies novel regulation of host signaling pathways. PLoS Pathog 9: e1003779.[Crossref]
  11. Molestina RE, El-Guendy N, Sinai AP, , 2008. Infection with Toxoplasma gondii results in dysregulation of the host cell cycle. Cell Microbiol 10: 11531165.[Crossref]
  12. Bottova I, Sauder U, Olivieri V, Hehl AB, Sonda S, , 2010. The P-glycoprotein inhibitor GF120918 modulates Ca2+-dependent processes and lipid metabolism in Toxoplasma gondii . PLoS One 5: e10062.[Crossref]
  13. Da SC, Da SE, Cruz MC, Chavrier P, Mortara RA, , 2009. ARF6, PI3-kinase and host cell actin cytoskeleton in Toxoplasma gondii cell invasion. Biochem Biophys Res Commun 378: 656661.[Crossref]
  14. Na RH, Zhu GH, Luo JX, Meng XJ, Cui L, Peng HJ, Chen XG, Gomez-Cambronero J, , 2013. Enzymatically active Rho and Rac small-GTPases are involved in the establishment of the vacuolar membrane after Toxoplasma gondii invasion of host cells. BMC Microbiol 13: 125.[Crossref]
  15. Treeck M, Sanders JL, Elias JE, Boothroyd JC, , 2011. The phosphoproteomes of Plasmodium falciparum and Toxoplasma gondii reveal unusual adaptations within and beyond the parasites' boundaries. Cell Host Microbe 10: 410419.[Crossref]
  16. Bodenmiller B, Malmstrom J, Gerrits B, Campbell D, Lam H, Schmidt A, Rinner O, Mueller LN, Shannon PT, Pedrioli PG, Panse C, Lee HK, Schlapbach R, Aebersold R, , 2007. PhosphoPep-a phosphoproteome resource for systems biology research in Drosophila Kc167 cells. Mol Syst Biol 3: 139.[Crossref]
  17. Villen J, Gygi SP, , 2008. The SCX/IMAC enrichment approach for global phosphorylation analysis by mass spectrometry. Nat Protoc 3: 16301638.[Crossref]
  18. Elias JE, Gygi SP, , 2007. Target-decoy search strategy for increased confidence in large-scale protein identifications by mass spectrometry. Nat Methods 4: 207214.[Crossref]
  19. Taus T, Kocher T, Pichler P, Paschke C, Schmidt A, Henrich C, Mechtler K, , 2011. Universal and confident phosphorylation site localization using phosphoRS. J Proteome Res 10: 53545362.[Crossref]
  20. Wu X, Tian L, Li J, Zhang Y, Han V, Li Y, Xu X, Li H, Chen X, Chen J, Jin W, Xie Y, Han J, Zhong CQ, , 2012. Investigation of receptor interacting protein (RIP3)-dependent protein phosphorylation by quantitative phosphoproteomics. Mol Cell Proteomics 11: 16401651.[Crossref]
  21. Graves JD, Krebs EG, , 1999. Protein phosphorylation and signal transduction. Pharmacol Ther 82: 111121.[Crossref]
  22. Marks F, , 2001. Protein phosphorylation: cellular functions and therapeutic perspectives. Tenth colloquium on cellular signal transduction: DKFZ Heidelberg, 9 February 2001. J Cancer Res Clin Oncol 127: 744750.
  23. Mundy J, Schneitz K, , 2002. Protein phosphorylation in and around signal transduction. Trends Plant Sci 7: 5455.[Crossref]
  24. Sawyer TK, Shakespeare WC, Wang Y, Sundaramoorthi R, Huang WS, Metcalf CR, Thomas M, Lawrence BM, Rozamus L, Noehre J, Zhu X, Narula S, Bohacek RS, Weigele M, Dalgarno DC, , 2005. Protein phosphorylation and signal transduction modulation: chemistry perspectives for small-molecule drug discovery. Med Chem 1: 293319.[Crossref]
  25. Ling YM, Shaw MH, Ayala C, Coppens I, Taylor GA, Ferguson DJ, Yap GS, , 2006. Vacuolar and plasma membrane stripping and autophagic elimination of Toxoplasma gondii in primed effector macrophages. J Exp Med 203: 20632071.[Crossref]
  26. Pawlowski N, Khaminets A, Hunn JP, Papic N, Schmidt A, Uthaiah RC, Lange R, Vopper G, Martens S, Wolf E, Howard JC, , 2011. The activation mechanism of Irga6, an interferon-inducible GTPase contributing to mouse resistance against Toxoplasma gondii . BMC Biol 9: 7.[Crossref]
  27. Wang Y, Weiss LM, Orlofsky A, , 2010. Coordinate control of host centrosome position, organelle distribution, and migratory response by Toxoplasma gondii via host mTORC2. J Biol Chem 285: 1561115618.[Crossref]
  28. Dvorak JA, Crane MS, , 1981. Vertebrate cell cycle modulates infection by protozoan parasites. Science 214: 10341036.[Crossref]
  29. Grimwood J, Mineo JR, Kasper LH, , 1996. Attachment of Toxoplasma gondii to host cells is host cell cycle dependent. Infect Immun 64: 40994104.
  30. Lavine MD, Arrizabalaga G, , 2009. Induction of mitotic S-phase of host and neighboring cells by Toxoplasma gondii enhances parasite invasion. Mol Biochem Parasitol 164: 9599.[Crossref]
  31. Williams GT, , 1994. Programmed cell death: a fundamental protective response to pathogens. Trends Microbiol 2: 463464.[Crossref]
  32. Green DR, , 2003. Overview: apoptotic signaling pathways in the immune system. Immunol Rev 193: 59.[Crossref]
  33. Duronio V, , 2008. The life of a cell: apoptosis regulation by the PI3K/PKB pathway. Biochem J 415: 333344.[Crossref]
  34. Quan JH, Cha GH, Zhou W, Chu JQ, Nishikawa Y, Lee YH, , 2013. Involvement of PI 3 kinase/Akt-dependent Bad phosphorylation in Toxoplasma gondii-mediated inhibition of host cell apoptosis. Exp Parasitol 133: 462471.[Crossref]
  35. Datta SR, Dudek H, Tao X, Masters S, Fu H, Gotoh Y, Greenberg ME, , 1997. Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell 91: 231241.[Crossref]
  36. Howells CC, Baumann WT, Samuels DC, Finkielstein CV, , 2011. The Bcl-2-associated death promoter (BAD) lowers the threshold at which the Bcl-2-interacting domain death agonist (BID) triggers mitochondria disintegration. J Theor Biol 271: 114123.[Crossref]
  37. Rombouts K, Lottini B, Caligiuri A, Liotta F, Mello T, Carloni V, Marra F, Pinzani M, , 2008. MARCKS is a downstream effector in platelet-derived growth factor-induced cell motility in activated human hepatic stellate cells. Exp Cell Res 314: 14441454.[Crossref]
  38. Thelen M, Rosen A, Nairn AC, Aderem A, , 1991. Regulation by phosphorylation of reversible association of a myristoylated protein kinase C substrate with the plasma membrane. Nature 351: 320322.[Crossref]
  39. Tinoco LW, Fraga JL, Anobom CD, Zolessi FR, Obal G, Toledo A, Pritsch O, Arruti C, , 2014. Structural characterization of a neuroblast-specific phosphorylated region of MARCKS. Biochim Biophys Acta 1844: 837849.[Crossref]
  40. Michaut MA, Williams CJ, Schultz RM, , 2005. Phosphorylated MARCKS: a novel centrosome component that also defines a peripheral subdomain of the cortical actin cap in mouse eggs. Dev Biol 280: 2637.[Crossref]
  41. Halonen SK, Weidner E, , 1994. Overcoating of Toxoplasma parasitophorous vacuoles with host cell vimentin type intermediate filaments. J Eukaryot Microbiol 41: 6571.[Crossref]
  42. Aziz A, Hess JF, Budamagunta MS, Voss JC, Fitzgerald PG, , 2010. Site-directed spin labeling and electron paramagnetic resonance determination of vimentin head domain structure. J Biol Chem 285: 1527815285.[Crossref]

Data & Media loading...

Supplementary Data

Supplemental Figure and Table

  • Received : 14 Nov 2016
  • Accepted : 27 Feb 2017

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error