The Moving Junction Protein RON8 Facilitates Firm Attachment and Host Cell Invasion in

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The apicomplexan moving junction (MJ) is a highly conserved structure formed during host cell entry that anchors the invading parasite to the host cell and serves as a molecular sieve of host membrane proteins that protects the parasitophorous vacuole from host lysosomal destruction. While recent work in Toxoplasma and Plasmodium has reinforced the composition of the MJ as an important association of rhoptry neck proteins (RONs) with micronemal AMA1, little is known of the precise role of RONs in the junction or how they are targeted to the neck subcompartment. We report the first functional analysis of a MJ/RON protein by disrupting RON8 in T. gondii. Parasites lacking RON8 are severely impaired in both attachment and invasion, indicating that RON8 enables the parasite to establish a firm clasp on the host cell and commit to invasion. The remaining junction components frequently drag in trails behind invading knockout parasites and illustrate a malformed complex without RON8. Complementation of Δron8 parasites restores invasion and reveals a processing event at the RON8 C-terminus. Replacement of an N-terminal region of RON8 with a mCherry reporter separates regions within RON8 that are necessary for rhoptry targeting and complex formation from those required for function during invasion. Finally, the invasion defects in Δron8 parasites seen in vitro translate to radically impaired virulence in infected mice, promoting a model in which RON8 has a crucial and unprecedented task in committing Toxoplasma to host cell entry.

Vyšlo v časopise: The Moving Junction Protein RON8 Facilitates Firm Attachment and Host Cell Invasion in. PLoS Pathog 7(3): e32767. doi:10.1371/journal.ppat.1002007
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002007

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1. BaumJRichardDHealerJRugMKrnajskiZ 2006 A conserved molecular motor drives cell invasion and gliding motility across malaria life cycle stages and other apicomplexan parasites. J Biol Chem 281 5197 5208

2. CowmanAFCrabbBS 2006 Invasion of red blood cells by malaria parasites. Cell 124 755 766

3. CarruthersVBoothroydJC 2007 Pulling together: an integrated model of Toxoplasma cell invasion. Curr Opin Microbiol 10 83 89

4. AikawaMMillerLHJohnsonJRabbegeJ 1978 Erythrocyte entry by malarial parasites. A moving junction between erythrocyte and parasite. J Cell Biol 77 72 82

5. Soldati-FavreD 2008 Molecular dissection of host cell invasion by the apicomplexans: the glideosome. Parasite 15 197 205

6. MordueDGDesaiNDustinMSibleyLD 1999 Invasion by Toxoplasma gondii establishes a moving junction that selectively excludes host cell plasma membrane proteins on the basis of their membrane anchoring. J Exp Med 190 1783 1792

7. CharronAJSibleyLD 2004 Molecular partitioning during host cell penetration by Toxoplasma gondii. Traffic 5 855 867

8. AlexanderDLMitalJWardGEBradleyPBoothroydJC 2005 Identification of the moving junction complex of Toxoplasma gondii: a collaboration between distinct secretory organelles. PLoS Pathog 1 e17

9. BesteiroSMichelinAPoncetJDubremetzJFLebrunM 2009 Export of a Toxoplasma gondii rhoptry neck protein complex at the host cell membrane to form the moving junction during invasion. PLoS Pathog 5 e1000309

10. LebrunMMichelinAEl HajjHPoncetJBradleyPJ 2005 The rhoptry neck protein RON4 re-localizes at the moving junction during Toxoplasma gondii invasion. Cell Microbiol 7 1823 1833

11. StraubKWChengSJSohnCSBradleyPJ 2009 Novel components of the Apicomplexan moving junction reveal conserved and coccidia-restricted elements. Cell Microbiol 11 590 603

12. BradleyPJWardCChengSJAlexanderDLCollerS 2005 Proteomic analysis of rhoptry organelles reveals many novel constituents for host-parasite interactions in Toxoplasma gondii. J Biol Chem 280 34245 34258

13. BoothroydJCDubremetzJF 2008 Kiss and spit: the dual roles of Toxoplasma rhoptries. Nat Rev Microbiol 6 79 88

14. GilbertLARavindranSTuretzkyJMBoothroydJCBradleyPJ 2007 Toxoplasma gondii targets a protein phosphatase 2C to the nuclei of infected host cells. Eukaryot Cell 6 73 83

15. SaeijJPCollerSBoyleJPJeromeMEWhiteMW 2007 Toxoplasma co-opts host gene expression by injection of a polymorphic kinase homologue. Nature 445 324 327

16. AlexanderDLArastu-KapurSDubremetzJFBoothroydJC 2006 Plasmodium falciparum AMA1 binds a rhoptry neck protein homologous to TgRON4, a component of the moving junction in Toxoplasma gondii. Eukaryot Cell 5 1169 1173

17. CaoJKanekoOThongkukiatkulATachibanaMOtsukiH 2009 Rhoptry neck protein RON2 forms a complex with microneme protein AMA1 in Plasmodium falciparum merozoites. Parasitol Int 58 29 35

18. NarumDLNguyenVZhangYGlenJShimpRL 2008 Identification and characterization of the Plasmodium yoelii PyP140/RON4 protein, an orthologue of Toxoplasma gondii RON4, whose cysteine-rich domain does not protect against lethal parasite challenge infection. Infect Immun 76 4876 4882

19. RichardDMacRaildCARiglarDTChanJAFoleyM 2010 Interaction between Plasmodium falciparum apical membrane antigen 1 and the rhoptry neck protein complex defines a key step in the erythrocyte invasion process of malaria parasites. J Biol Chem 285 14815 14822

20. TrigliaTThamWHHodderACowmanAF 2009 Reticulocyte binding protein homologues are key adhesins during erythrocyte invasion by Plasmodium falciparum. Cell Microbiol 11 1671 1687

21. ProellocksNICoppelRLWallerKL 2010 Dissecting the apicomplexan rhoptry neck proteins. Trends Parasitol 26 297 304

22. MitalJMeissnerMSoldatiDWardGE 2005 Conditional expression of Toxoplasma gondii apical membrane antigen-1 (TgAMA1) demonstrates that TgAMA1 plays a critical role in host cell invasion. Mol Biol Cell 16 4341 4349

23. FoxBARistucciaJGGigleyJPBzikDJ 2009 Efficient gene replacements in Toxoplasma gondii strains deficient for nonhomologous end joining. Eukaryot Cell 8 520 529

24. HuynhMHCarruthersVB 2009 Tagging of endogenous genes in a Toxoplasma gondii strain lacking Ku80. Eukaryot Cell 8 530 539

25. BeckJRRodriguez-FernandezIACruz de LeonJHuynhMHCarruthersVB 2010 A Novel Family of Toxoplasma IMC Proteins Displays a Hierarchical Organization and Functions in Coordinating Parasite Division. PLoS Pathog 6. pii e1001094

26. CareyKLJongcoAMKimKWardGE 2004 The Toxoplasma gondii rhoptry protein ROP4 is secreted into the parasitophorous vacuole and becomes phosphorylated in infected cells. Eukaryot Cell 3 1320 1330

27. HuynhMHRabenauKEHarperJMBeattyWLSibleyLD 2003 Rapid invasion of host cells by Toxoplasma requires secretion of the MIC2-M2AP adhesive protein complex. Embo J 22 2082 2090

28. CarruthersVBSibleyLD 1997 Sequential protein secretion from three distinct organelles of Toxoplasma gondii accompanies invasion of human fibroblasts. Eur J Cell Biol 73 114 123

29. HakanssonSCharronAJSibleyLD 2001 Toxoplasma evacuoles: a two-step process of secretion and fusion forms the parasitophorous vacuole. Embo J 20 3132 3144

30. LalKBromleyEOakesRPrietoJHSandersonSJ 2009 Proteomic comparison of four Eimeria tenella life-cycle stages: unsporulated oocyst, sporulated oocyst, sporozoite and second-generation merozoite. Proteomics 9 4566 4576

31. Tufet-BayonaMJanseCJKhanSMWatersAPSindenRE 2009 Localisation and timing of expression of putative Plasmodium berghei rhoptry proteins in merozoites and sporozoites. Mol Biochem Parasitol 166 22 31

32. GonzalezVCombeADavidVMalmquistNADelormeV 2009 Host cell entry by apicomplexa parasites requires actin polymerization in the host cell. Cell Host Microbe 5 259 272

33. MillerSAThathyVAjiokaJWBlackmanMJKimK 2003 TgSUB2 is a Toxoplasma gondii rhoptry organelle processing proteinase. Mol Microbiol 49 883 894

34. ReissMViebigNBrechtSFourmauxMNSoeteM 2001 Identification and characterization of an escorter for two secretory adhesins in Toxoplasma gondii. J Cell Biol 152 563 578

35. RichardDKatsLMLangerCBlackCGMitriK 2009 Identification of rhoptry trafficking determinants and evidence for a novel sorting mechanism in the malaria parasite Plasmodium falciparum. PLoS Pathog 5 e1000328

36. SaourosSEdwards-JonesBReissMSawmynadenKCotaE 2005 A novel galectin-like domain from Toxoplasma gondii micronemal protein 1 assists the folding, assembly, and transport of a cell adhesion complex. J Biol Chem 280 38583 38591

37. MorahanBJSallmannGBHuestisRDubljevicVWallerKL 2009 Plasmodium falciparum: genetic and immunogenic characterisation of the rhoptry neck protein PfRON4. Exp Parasitol 122 280 288

38. DonaldRGCarterDUllmanBRoosDS 1996 Insertional tagging, cloning, and expression of the Toxoplasma gondii hypoxanthine-xanthine-guanine phosphoribosyltransferase gene. Use as a selectable marker for stable transformation. J Biol Chem 271 14010 14019

39. SadakATaghyZFortierBDubremetzJF 1988 Characterization of a family of rhoptry proteins of Toxoplasma gondii. Mol Biochem Parasitol 29 203 211

40. TuretzkyJMChuDKHajagosBEBradleyPJ 2010 Processing and secretion of ROP13: A unique Toxoplasma effector protein. Int J Parasitol 40 1037 44

41. RomeMEBeckJRTuretzkyJMWebsterPBradleyPJ 2008 Intervacuolar transport and unique topology of GRA14, a novel dense granule protein in Toxoplasma gondii. Infect Immun 76 4865 4875

42. BradleyPJHsiehCLBoothroydJC 2002 Unprocessed Toxoplasma ROP1 is effectively targeted and secreted into the nascent parasitophorous vacuole. Mol Biochem Parasitol 125 189 193

43. KarasovAOBoothroydJCArrizabalagaG 2005 Identification and disruption of a rhoptry-localized homologue of sodium hydrogen exchangers in Toxoplasma gondii. Int J Parasitol 35 285 291

44. DobrowolskiJMSibleyLD 1996 Toxoplasma invasion of mammalian cells is powered by the actin cytoskeleton of the parasite. Cell 84 933 939