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Micronemal protein 13 contributes to the optimal growth of Toxoplasma gondii under stress conditions

  • Shu Ye
  • Ningbo Xia
  • Pengfei Zhao
  • Jichao Yang
  • Yanqin Zhou
  • Bang Shen
  • Junlong Zhao
Protozoology - Original Paper
  • 131 Downloads

Abstract

Toxoplasma gondii is a ubiquitous parasitic protozoan infecting humans and a wide variety of animals. Fast-replicating tachyzoites during acute infection and slowly growing bradyzoites during chronic infection are the two basic forms of T. gondii in intermediate hosts. Interconversion between the two contributes to the transmission and pathogenesis of this parasite. Secretory micronemal proteins are thought to mediate interactions with host cells and facilitate parasite invasion, therefore the majority of them are highly expressed in tachyzoites. Micronemal protein 13 (MIC13) is unique in that its expression is low in tachyzoites and is upregulated under bradyzoite-inducing conditions. Previous attempts to disrupt this gene were not successful, implying that it may play critical roles during parasite growth. However, in this study, MIC13 was successfully disrupted in type 1 strain RH and type 2 strain ME49 using CRISPR/Cas9-mediated gene disruption techniques. Consistent with its low expression in tachyzoites and increased expression under stress or bradyzoite-inducing conditions, MIC13-inactivated mutants displayed normal growth, host cell invasion, intracellular replication, and egress, as well as acute virulence at the tachyzoite stage. However, under stress conditions, such as high pH or oxygen limitation, MIC13-disrupted parasites showed significantly slower growth rates compared to the parental strains, suggesting that it is required for optimal parasite growth under bradyzoite-inducing or stress conditions. This is the first micronemal protein reported to have such expression pattern and function modes, which expands our understanding of the diverse functions of micronemal proteins.

Keywords

MIC13 Bradyzoite Stress Toxoplasma gondii Micronemal protein 

Notes

Funding information

This work was supported the National Key Research and Development Program of China (Grant no. 2017YFD0500402), the National Basic Science Research Program (973 program) of China (Grant no. 2015CB150300), and the Natural Science Foundation of Hubei Province (Project 2017CFA020). The funders had no role in the study design, data collection and analysis, preparation of the manuscript, or decision to submit the work for publication.

Compliance with ethical standards

All animal experiments were approved by the Ethical Committee of Huazhong Agricultural University (permit no. HZAUMO-2017-023).

Competing interests

The authors declare they have no conflict of interest.

References

  1. Bargieri DY, Andenmatten N, Lagal V, Thiberge S, Whitelaw JA, Tardieux I, Meissner M, Ménard R (2013) Apical membrane antigen 1 mediates apicomplexan parasite attachment but is dispensable for host cell invasion. Nat Commun 4:2552CrossRefGoogle Scholar
  2. Blumenschein TM et al (2007) Atomic resolution insight into host cell recognition by toxoplasma gondii. EMBO J 26:2808–2820CrossRefGoogle Scholar
  3. Brecht S, Carruthers VB, Ferguson DJP, Giddings OK, Wang G, Jäkle U, Harper JM, Sibley LD, Soldati D (2001) The toxoplasma micronemal protein MIC4 is an adhesin composed of six conserved apple domains. J Biol Chem 276:4119–4127CrossRefGoogle Scholar
  4. Carruthers V, Boothroyd JC (2007) Pulling together: an integrated model of Toxoplasma cell invasion. Curr Opin Microbiol 10:83–89CrossRefGoogle Scholar
  5. Carruthers VB, Tomley FM (2008) Microneme Proteins in Apicomplexans. In: Microneme proteins in apicomplexans. Springer, New YorkCrossRefGoogle Scholar
  6. Cérède O, Dubremetz JF, Soête M, Deslée D, Vial H, Bout D, Lebrun M (2005) Synergistic role of micronemal proteins in Toxoplasma gondii virulence. J Exp Med 201:453–463CrossRefGoogle Scholar
  7. Daher W, Plattner F, Carlier MF, Soldatifavre D (2010) Concerted action of two formins in gliding motility and host cell invasion by Toxoplasma gondii. PLoS Pathog 6:e1001132CrossRefGoogle Scholar
  8. Dubey JP, Hill DE, Rozeboom DW, Rajendran C, Choudhary S, Ferreira LR, Kwok OCH, Su C (2012) High prevalence and genotypes of Toxoplasma gondii isolated from organic pigs in northern USA. Vet Parasitol 188:14–18CrossRefGoogle Scholar
  9. Elmore SA, Jones JL, Conrad PA, Patton S, Lindsay DS, Dubey JP (2010) Toxoplasma gondii: epidemiology, feline clinical aspects, and prevention. Trends Parasitol 26:190–196.  https://doi.org/10.1016/j.pt.2010.01.009 CrossRefGoogle Scholar
  10. Ferreira da Silva Mda F, Barbosa HS, Gross U, Luder CG (2008) Stress-related and spontaneous stage differentiation of toxoplasma gondii. Mol BioSyst 4:824–834.  https://doi.org/10.1039/b800520f CrossRefGoogle Scholar
  11. Frénal K, Dubremetz JF, Lebrun M, Soldatifavre D (2017) Gliding motility powers invasion and egress in Apicomplexa. Nat Rev Microbiol 15:645–660CrossRefGoogle Scholar
  12. Friedrich N, Santos JM, Liu Y, Palma AS, Leon E, Saouros S, Kiso M, Blackman MJ, Matthews S, Feizi T, Soldati-Favre D (2010) Members of a novel protein family containing microneme adhesive repeat domains act as sialic acid-binding lectins during host cell invasion by apicomplexan parasites. J Biol Chem 285:2064–2076CrossRefGoogle Scholar
  13. Fritz HM, Buchholz KR, Chen X, Durbinjohnson B, Rocke DM, Conrad PA, Boothroyd JC (2012) Transcriptomic analysis of toxoplasma development reveals many novel functions and structures specific to sporozoites and oocysts. PLoS One 7:e29998CrossRefGoogle Scholar
  14. Hirdes W, Davis DW, Eisenlohr BN (2012) Prevalence and genotypes of Toxoplasma gondii in pork from retail meat stores in Eastern China. Int J Food Microbiol 157:393CrossRefGoogle Scholar
  15. Hunter CA, Sibley LD (2012) Modulation of innate immunity by Toxoplasma gondii virulence effectors. Nat Rev Microbiol 10:766–778CrossRefGoogle Scholar
  16. Huynh MH, Carruthers VB (2006) Toxoplasma MIC2 is a major determinant of invasion and virulence. PLoS Pathog 2:e84CrossRefGoogle Scholar
  17. Huynh MH, Rabenau KE, Harper JM, Beatty WL, Sibley LD, Carruthers VB (2003) Rapid invasion of host cells by Toxoplasma requires secretion of the MIC2-M2AP adhesive protein complex. EMBO J 22:2082–2090CrossRefGoogle Scholar
  18. Jr SW, Jeffers V (2012) Mechanisms of Toxoplasma gondii persistence and latency. FEMS Microbiol Rev 36:717–733CrossRefGoogle Scholar
  19. Kessler H, Herm-Götz A, Hegge S, Rauch M, Soldati-Favre D, Frischknecht F, Meissner M (2008) Microneme protein 8--a new essential invasion factor in Toxoplasma gondii. J Cell Sci 121:947–956CrossRefGoogle Scholar
  20. Krishnamurthy S, Deng B, del Rio R, Buchholz KR, Treeck M, Urban S, Boothroyd J, Lam YW, Ward GE (2016) Not a simple tether: binding of toxoplasma gondii AMA1 to RON2 during invasion protects AMA1 from rhomboid-mediated cleavage and leads to dephosphorylation of its cytosolic tail. mBio 7(5):e00754-16Google Scholar
  21. Lyons RE, Mcleod R, Roberts CW (2002) Toxoplasma gondii tachyzoite-bradyzoite interconversion. Trends Parasitol 18:198–201CrossRefGoogle Scholar
  22. Matthias R et al (2001) Identification and characterization of an escorter for two secretory adhesins in Toxoplasma gondii. J Cell Biol 152:563–578CrossRefGoogle Scholar
  23. Meissner M, Reiss M, Viebig N, Carruthers VB, Toursel C, Tomavo S, Ajioka JW, Soldati D (2002) A family of transmembrane microneme proteins of Toxoplasma gondii contain EGF-like domains and function as escorters. J Cell Sci 115:563–574Google Scholar
  24. Montoya JG, Liesenfeld O (2004) Toxoplasmosis. Lancet 363:1965–1976CrossRefGoogle Scholar
  25. Pittman KJ, Aliota MT, Knoll LJ (2014) Dual transcriptional profiling of mice and Toxoplasma gondii during acute and chronic infection. BMC Genomics 15(1(2014-09-20) 15):806CrossRefGoogle Scholar
  26. Radke JB, Lucas O, de Silva EK, Ma Y, Sullivan WJ Jr, Weiss LM, Llinas M, White MW (2013) ApiAP2 transcription factor restricts development of the Toxoplasma tissue cyst. Proc Natl Acad Sci U S A 110:6871–6876CrossRefGoogle Scholar
  27. Radke JB, Worth D, Hong D (2018) Transcriptional repression by ApiAP2 factors is central to chronic toxoplasmosis. PLoS Pathog 14:e1007035.  https://doi.org/10.1371/journal.ppat.1007035 CrossRefGoogle Scholar
  28. Remington JS, Cavanaugh EN (1965) Isolation of the encysted form of Toxoplasma gondii from human skeletal muscle and brain. N Engl J Med 273:1308–1310CrossRefGoogle Scholar
  29. Selseleh M, Modarressi MH, Mohebali M, Shojaee S, Eshragian MR, Selseleh M, Azizi E, Keshavarz H (2012) Real-time RT-PCR on SAG1 and BAG1 gene expression during stage conversion in immunosuppressed mice infected with Toxoplasma gondii Tehran strain. Korean J Parasitol 50:199–205CrossRefGoogle Scholar
  30. Shen B, Sibley LD (2014) Toxoplasma aldolase is required for metabolism but dispensable for host-cell invasion. Proc Natl Acad Sci U S A 111:3567–3572.  https://doi.org/10.1073/pnas.1315156111 CrossRefGoogle Scholar
  31. Shen B, Brown KM, Lee TD, Sibley LD (2014) Efficient gene disruption in diverse strains of Toxoplasma gondii using CRISPR/CAS9. Mbio 5:01114–01114Google Scholar
  32. Skariah S, McIntyre MK, Mordue DG (2010) Toxoplasma gondii: determinants of tachyzoite to bradyzoite conversion. Parasitol Res 107:253–260.  https://doi.org/10.1007/s00436-010-1899-6 CrossRefGoogle Scholar
  33. Soete M, Fortier B, Camus D, Dubremetz JF (1993) Toxoplasma gondii: kinetics of bradyzoite-tachyzoite interconversion in vitro. Exp Parasitol 76:259–264.  https://doi.org/10.1006/expr.1993.1031 CrossRefGoogle Scholar
  34. Ueno A, Dautu G, Munyaka B, Carmen G, Kobayashi Y, Igarashi M (2009) Toxoplasma gondii : identification and characterization of bradyzoite-specific deoxyribose phosphate aldolase-like gene ( Tg DPA). Exp Parasitol 121:55–63CrossRefGoogle Scholar
  35. Walker R, Gissot M, Croken MM, Huot L, Hot D, Kim K, Tomavo S (2013) The Toxoplasma nuclear factor TgAP2XI-4 controls bradyzoite gene expression and cyst formation. Mol Microbiol 87:641–655CrossRefGoogle Scholar
  36. White MW, Radke JR, Radke JB (2014) Toxoplasma development - turn the switch on or off? Cell Microbiol 16:466–472CrossRefGoogle Scholar
  37. Xia N, Yang J, Ye S, Zhang L, Zhou Y, Zhao J, David Sibley L, Shen B (2018) Functional analysis of Toxoplasma lactate dehydrogenases suggests critical roles of lactate fermentation for parasite growth in vivo. Cell Microbiol 20:e12794CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Agricultural MicrobiologyHuazhong Agricultural UniversityWuhanPeople’s Republic of China
  2. 2.Key Laboratory of Preventive Medicine in Hubei ProvinceWuhanPeople’s Republic of China
  3. 3.Hubei Cooperative Innovation Center for Sustainable Pig ProductionWuhanPeople’s Republic of China

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