The Cyst Wall Protein CST1 Is Critical for Cyst Wall Integrity and Promotes Bradyzoite Persistence
Toxoplasma gondii infects up to one third of the world's population. A key to the success of T. gondii as a parasite is its ability to persist for the life of its host as bradyzoites within tissue cysts. The glycosylated cyst wall is the key structural feature that facilitates persistence and oral transmission of this parasite. Because most of the antibodies and reagents that recognize the cyst wall recognize carbohydrates, identification of the components of the cyst wall has been technically challenging. We have identified CST1 (TGME49_064660) as a 250 kDa SRS (SAG1 related sequence) domain protein with a large mucin-like domain. CST1 is responsible for the Dolichos biflorus Agglutinin (DBA) lectin binding characteristic of T. gondii cysts. Deletion of CST1 results in reduced cyst number and a fragile brain cyst phenotype characterized by a thinning and disruption of the underlying region of the cyst wall. These defects are reversed by complementation of CST1. Additional complementation experiments demonstrate that the CST1-mucin domain is necessary for the formation of a normal cyst wall structure, the ability of the cyst to resist mechanical stress, and binding of DBA to the cyst wall. RNA-seq transcriptome analysis demonstrated dysregulation of bradyzoite genes within the various cst1 mutants. These results indicate that CST1 functions as a key structural component that confers essential sturdiness to the T. gondii tissue cyst critical for persistence of bradyzoite forms.
Vyšlo v časopise:
The Cyst Wall Protein CST1 Is Critical for Cyst Wall Integrity and Promotes Bradyzoite Persistence. PLoS Pathog 9(12): e32767. doi:10.1371/journal.ppat.1003823
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1003823
Souhrn
Toxoplasma gondii infects up to one third of the world's population. A key to the success of T. gondii as a parasite is its ability to persist for the life of its host as bradyzoites within tissue cysts. The glycosylated cyst wall is the key structural feature that facilitates persistence and oral transmission of this parasite. Because most of the antibodies and reagents that recognize the cyst wall recognize carbohydrates, identification of the components of the cyst wall has been technically challenging. We have identified CST1 (TGME49_064660) as a 250 kDa SRS (SAG1 related sequence) domain protein with a large mucin-like domain. CST1 is responsible for the Dolichos biflorus Agglutinin (DBA) lectin binding characteristic of T. gondii cysts. Deletion of CST1 results in reduced cyst number and a fragile brain cyst phenotype characterized by a thinning and disruption of the underlying region of the cyst wall. These defects are reversed by complementation of CST1. Additional complementation experiments demonstrate that the CST1-mucin domain is necessary for the formation of a normal cyst wall structure, the ability of the cyst to resist mechanical stress, and binding of DBA to the cyst wall. RNA-seq transcriptome analysis demonstrated dysregulation of bradyzoite genes within the various cst1 mutants. These results indicate that CST1 functions as a key structural component that confers essential sturdiness to the T. gondii tissue cyst critical for persistence of bradyzoite forms.
Zdroje
1. TenterAM, HeckerothAR, WeissLM (2000) Toxoplasma gondii: from animals to humans. International Journal for Parasitology 30: 1217–1258.
2. PappasG, RoussosN, FalagasME (2009) Toxoplasmosis snapshots: global status of Toxoplasma gondii seroprevalence and implications for pregnancy and congenital toxoplasmosis. International Journal for Parasitology 39: 1385–1394.
3. FergusonDJ, HutchisonWM (1987) The host-parasite relationship of Toxoplasma gondii in the brains of chronically infected mice. Virchows Archiv A, Pathological anatomy and histopathology 411: 39–43.
4. HillD, DubeyJP (2002) Toxoplasma gondii: transmission, diagnosis and prevention. Clinical Microbiology and Infection 8: 634–640.
5. FergusonDJ, HutchisonWM, PettersenE (1989) Tissue cyst rupture in mice chronically infected with Toxoplasma gondii. An immunocytochemical and ultrastructural study. Parasitology Research 75: 599–603.
6. DubeyJP, LindsayDS, SpeerCA (1998) Structures of Toxoplasma gondii Tachyzoites, Bradyzoites, and Sporozoites and Biology and Development of Tissue Cysts Clinical Microbiology Reviews. 11: 267–299.
7. SimsTA, HayJ, TalbotIC (1988) Host-parasite relationship in the brains of mice with congenital toxoplasmosis. The Journal of Pathology 156: 255–261.
8. BoothroydJC, BlackM, BonnefoyS, HehlA, KnollLJ, et al. (1997) Genetic and biochemical analysis of development in Toxoplasma gondii. Philosophical transactions of the Royal Society of London Series B, Biological sciences 352: 1347–1354.
9. WeissLM, KimK (2000) The development and biology of bradyzoites of Toxoplasma gondii. Frontiers in Bioscience 5: D391–405.
10. ZhangYW, HalonenSK, MaYF, WittnerM, WeissLM (2001) Initial characterization of CST1, a Toxoplasma gondii cyst wall glycoprotein. Infection and Immunity 69: 501–507.
11. WeissLM, LaPlaceD, TanowitzHB, WittnerM (1992) Identification of Toxoplasma gondii bradyzoite-specific monoclonal antibodies. The Journal of Infectious Diseases 166: 213–215.
12. Weiss LM, Kim K (2007) Bradyzoite development. In: Weiss LM, Kim K, editors. Toxoplasma gondii: The Model Apicomplexan. Elsevier Ltd. pp. 341–366.
13. BuchholzKR, FritzHM, ChenX, Durbin-JohnsonB, RockeDM, et al. (2011) Identification of tissue cyst wall components by transcriptome analysis of in vivo and in vitro Toxoplasma gondii bradyzoites. Eukaryotic Cell 10: 1637–1647.
14. WasmuthJD, PszennyV, HaileS, JansenEM, GastAT, et al. (2012) Integrated Bioinformatic and Targeted Deletion Analyses of the SRS Gene Superfamily Identify SRS29C as a Negative Regulator of Toxoplasma Virulence. mBio 3: e00321–12.
15. PetersenTN, BrunakS, Von HeijneG, NielsenH (2011) SignalP 4.0: discriminating signal peptides from transmembrane regions. Nature Methods 8: 785–786.
16. JuleniusK, GuptaR (2005) Prediction, conservation analysis, and structural characterization of mammalian mucin-type O-glycosylation sites. Glycobiology 15: 153–164.
17. EtzlerME, KabatEA (1970) Purification and characterization of a lectin (plant hemagglutinin) with blood group A specificity from Dolichos biflorus. Biochemistry 9: 869–877.
18. FoxBA, FallaA, RommereimLM, TomitaT, GigleyJP, et al. (2011) Type II Toxoplasma gondii KU80 knockout strains enable functional analysis of genes required for cyst development and latent infection. Eukaryotic Cell 10: 1193–1206.
19. CornelissenAWCA, OverdulveJP, HoenderboomJM (1981) Separation of Isospora (Toxoplasma) gondii cysts and cystozoites from mouse brain tissue by continuous density - gradient centrifugation. Parasitology 83: 103.
20. BarnesDA, BonninA, HuangJX, GoussetL, WuJ, et al. (1998) A novel multi-domain mucin-like glycoprotein of Cryptosporidium parvum mediates invasion. Molecular and Biochemical Parasitology 96: 93–110.
21. ChatterjeeA, BanerjeeS, SteffenM, O'ConnorRM, WardHD, et al. (2010) Evidence for mucin-like glycoproteins that tether sporozoites of Cryptosporidium parvum to the inner surface of the oocyst wall. Eukaryotic Cell 9: 84–96.
22. MortaraRa, Da SilvaS, AraguthMF, BlancoSa, YoshidaN (1992) Polymorphism of the 35- and 50-kilodalton surface glycoconjugates of Trypanosoma cruzi metacyclic trypomastigotes. Infection and Immunity 60: 4673–4678.
23. BuscagliaCa, CampoVa, FraschACC, Di NoiaJM (2006) Trypanosoma cruzi surface mucins: host-dependent coat diversity. Nature Reviews Microbiology 4: 229–236.
24. McFaddenDC, TomavoS, BerryEA, BoothroydJC (2000) Characterization of cytochrome b from Toxoplasma gondii and Qo domain mutations as a mechanism of atovaquone-resistance. Molecular and Biochemical Parasitology 108: 1–12.
25. CraverMPJ, RooneyPJ, KnollLJ (2010) Isolation of Toxoplasma gondii development mutants identifies a potential proteophosphogylcan that enhances cyst wall formation. Molecular and Biochemical Parasitology 169: 120–123.
26. BuchholzKR, BowyerPW, BoothroydJC (2013) Bradyzoite Pseudokinase 1 Is Crucial for Efficient Oral Infectivity of the Toxoplasma gondii Tissue Cyst. Eukaryotic Cell 12: 399–410.
27. FergusonDJJP (2004) Use of molecular and ultrastructural markers to evaluate stage conversion of Toxoplasma gondii in both the intermediate and definitive host. International Journal for Parasitology 34: 347–360.
28. Stwora-WojczykMM, DzierszinskiF, RoosDS, SpitalnikSL, WojczykBS (2004) Functional characterization of a novel Toxoplasma gondii glycosyltransferase: UDP-N-acetyl-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase-T3. Archives of Biochemistry and Biophysics 426: 231–240.
29. CaffaroCE, KoshyAa, LiuL, ZeinerGM, HirschbergCB, et al. (2013) A Nucleotide Sugar Transporter Involved in Glycosylation of the Toxoplasma Tissue Cyst Wall Is Required for Efficient Persistence of Bradyzoites. PLoS Pathogens 9: e1003331.
30. BrownC (1990) Class I MHC genes and CD8+ T cells determine cyst number in Toxoplasma gondii infection. The Journal of Immunology 145: 3438–3441.
31. Madrid-AlisteCJ, DybasJM, AngelettiRH, WeissLM, KimK, et al. (2009) EPIC-DB: a proteomics database for studying Apicomplexan organisms. BMC Genomics 10: 38.
32. ZhangYW, KimK, MaYF, WittnerM, TanowitzHB, et al. (1999) Disruption of the Toxoplasma gondii bradyzoite-specific gene BAG1 decreases in vivo cyst formation. Molecular Microbiology 31: 691–701.
33. PfefferkornER, PfefferkornLC (1977) Toxoplasma gondii: specific labeling of nucleic acids of intracellular parasites in Lesch-Nyhan cells. Experimental Parasitology 41: 95–104.
34. SchwartzmanJD, PfefferkornER (1981) Pyrimidine synthesis by intracellular Toxoplasma gondii. The Journal of Parasitology 67: 150–158.
Štítky
Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
PLOS Pathogens
2013 Číslo 12
- Očkování proti virové hemoragické horečce Ebola experimentální vakcínou rVSVDG-ZEBOV-GP
- Parazitičtí červi v terapii Crohnovy choroby a dalších zánětlivých autoimunitních onemocnění
- Koronavirus hýbe světem: Víte jak se chránit a jak postupovat v případě podezření?
Najčítanejšie v tomto čísle
- Influence of Mast Cells on Dengue Protective Immunity and Immune Pathology
- Myeloid Dendritic Cells Induce HIV-1 Latency in Non-proliferating CD4 T Cells
- Host Defense via Symbiosis in
- Coronaviruses as DNA Wannabes: A New Model for the Regulation of RNA Virus Replication Fidelity