Obligate intracellular Apicomplexa parasites share a unique invasion mechanism involving a tight interaction between the host cell and the parasite surfaces called the moving junction (MJ). The MJ, which is the anchoring structure for the invasion process, is formed by secretion of a macromolecular complex (RON2/4/5/8), derived from secretory organelles called rhoptries, into the host cell membrane. AMA1, a protein secreted from micronemes and associated with the parasite surface during invasion, has been shown in vitro to bind the MJ complex through a direct association with RON2. Here we show that RON2 is inserted as an integral membrane protein in the host cell and, using several interaction assays with native or recombinant proteins, we define the region that binds AMA1. Our studies were performed both in Toxoplasma gondii and Plasmodium falciparum and although AMA1 and RON2 proteins have diverged between Apicomplexa species, we show an intra-species conservation of their interaction. More importantly, invasion inhibition assays using recombinant proteins demonstrate that the RON2-AMA1 interaction is crucial for both T. gondii and P. falciparum entry into their host cells. This work provides the first evidence that AMA1 uses the rhoptry neck protein RON2 as a receptor to promote invasion by Apicomplexa parasites.
Vyšlo v časopise: The RON2-AMA1 Interaction is a Critical Step in Moving Junction-Dependent Invasion by Apicomplexan Parasites. PLoS Pathog 7(2): e32767. doi:10.1371/journal.ppat.1001276 Kategorie: Research Article prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1001276
1. AikawaM
MillerLH
JohnsonJ
RabbegeJ
1978
Erythrocyte entry by malarial parasites. A moving junction between erythrocyte and parasite.
J Cell Biol
77
72
82
2. AlexanderDL
MitalJ
WardGE
BradleyP
BoothroydJC
2005
Identification of the Moving Junction Complex of Toxoplasma gondii: A Collaboration between Distinct Secretory Organelles.
PLoS Pathog
1
e17
3. LebrunM
MichelinA
El HajjH
PoncetJ
BradleyPJ
The rhoptry neck protein RON4 relocalizes at the moving junction during Toxoplasma gondii invasion.
Cell Microbiol
7
1823
1833
4. StraubKW
ChengSJ
SohnCS
2009
Novel components of the Apicomplexan moving junction reveal conserved and coccidia-restricted elements.
11
590
603
5. BesteiroS
DubremetzJF
LebrunM
Export of a Toxoplasma gondii rhoptry neck protein complex at the host cell membrane to form the moving junction during invasion.
Plos Pathogen
5
e1000309
6. DonahueCG
CarruthersVB
GilkSD
2000
The Toxoplasma homolog of Plasmodium apical membrane antigen-1 (AMA-1) is a microneme protein secreted in response to elevated intracellular calcium levels.
Mol Biochem Parasitol
111
15
30
7. HehlAB
LekutisC
GriggME
Toxoplasma gondii homologue of plasmodium apical membrane antigen 1 is involved in invasion of host cells.
Infect Immun
68
7078
7086
8. CarruthersVB
SibleyLD
1997
Sequential protein secretion from three distinct organelles of Toxoplasma gondii accompanies invasion of human fibroblasts.
Eur J Cell Biol
73
114
123
9. DeansJA
AldersonT
ThomasAW
MitchellGH
LennoxES
1982
Rat monoclonal antibodies which inhibit the in vitro multiplication of Plasmodium knowlesi.
Clin Exp Immunol
49
297
309
10. MitalJ
MeissnerM
SoldatiD
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
11. TrigliaT
HealerJ
CaruanaSR
HodderAN
AndersRF
Apical membrane antigen 1 plays a central role in erythrocyte invasion by Plasmodium species.
Mol Microbiol
38
706
718
12. HodderAN
CrewtherPE
2001
Specificity of the protective antibody response to apical membrane antigen 1.
69
3286
3294
13. RemarqueEJ
FaberBW
KockenCH
2008
Apical membrane antigen 1: a malaria vaccine candidate in review.
Trends Parasitol
24
74
84
14. CollinsCR
Withers-MartinezC
HackettF
BlackmanMJ
An inhibitory antibody blocks interactions between components of the malarial invasion machinery.
e1000273
15. CaoJ
KanekoO
ThongkukiatkulA
TachibanaM
OtsukiH
Rhoptry neck protein RON2 forms a complex with microneme protein AMA1 in Plasmodium falciparum merozoites.
Parasitol Int
58
29
35
16. NarumDL
NguyenV
ZhangY
GlenJ
ShimpRL
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.
76
4876
4882
17. KockenCH
DubbeldMA
van der WelA
2002
High-level expression of the malaria blood-stage vaccine candidate Plasmodium falciparum apical membrane antigen 1 and induction of antibodies that inhibit erythrocyte invasion.
70
4471
4476
18. PizarroJC
Vulliez-Le NormandB
Chesne-SeckML
CollinsCR
Crystal structure of the malaria vaccine candidate apical membrane antigen 1.
Science
308
408
411
19. BaiT
BeckerM
GuptaA
StrikeP
MurphyVJ
Structure of AMA1 from Plasmodium falciparum reveals a clustering of polymorphisms that surround a conserved hydrophobic pocket.
Proc Natl Acad Sci U S A
102
12736
12741
20. CrawfordJ
TonkinML
GrujicO
BoulangerMJ
2010
Structural characterization of apical membrane antigen 1 (AMA1) from Toxoplasma gondii.
J Biol Chem
285
15644
15652
21. ColeyAM
ParisiK
MasciantonioR
HoeckJ
CaseyJL
2006
The most polymorphic residue on Plasmodium falciparum apical membrane antigen 1 determines binding of an invasion-inhibitory antibody.
2628
2636
22. HendersonKA
StreltsovVA
ColeyAM
DolezalO
HudsonPJ
2007
Structure of an IgNAR-AMA1 complex: targeting a conserved hydrophobic cleft broadens malarial strain recognition.
Structure
1452
1466
23. RichardD
MacRaildCA
RiglarDT
ChanJA
FoleyM
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.
14815
14822
24. CeredeO
BoutD
The Toxoplasma gondii protein MIC3 requires pro-peptide cleavage and dimerization to function as adhesin.
Embo J
21
2526
2536
25. KesslerH
Herm-GotzA
HeggeS
RauchM
Soldati-FavreD
Microneme protein 8–a new essential invasion factor in Toxoplasma gondii.
J Cell Sci
121
947
956
26. IgonetS
FaureG
RiottotMM
Cross-reactivity studies of an anti-Plasmodium vivax apical membrane antigen 1 monoclonal antibody: binding and structural characterisation.
J Mol Biol
366
1523
1537
27. Chesne-SeckML
PizarroJC
Structural comparison of apical membrane antigen 1 orthologues and paralogues in apicomplexan parasites.
144
55
67
28. KusiKA
RemarqueEJ
Humoral immune response to mixed PfAMA1 alleles; multivalent PfAMA1 vaccines induce broad specificity.
PLoS One
4
e8110
29. KockenCH
Van Der WelA
PronkJT
WatersAP
1999
High-level expression of Plasmodium vivax apical membrane antigen 1 (AMA-1) in Pichia pastoris: strong immunogenicity in Macaca mulatta immunized with P. vivax AMA-1 and adjuvant SBAS2.
43
30. HarrisKS
SaboJK
Binding hot spot for invasion inhibitory molecules on Plasmodium falciparum apical membrane antigen 1.
6981
6989
31. DonaldRG
RoosDS
1998
Gene knock-outs and allelic replacements in Toxoplasma gondii: HXGPRT as a selectable marker for hit-and-run mutagenesis.
91
295
305
32. JensenJB
TragerW
1977
Plasmodium falciparum in culture: use of outdated erthrocytes and description of the candle jar method.
J Parasitol
63
883
886
33. KafsackBF
BeckersC
2004
Synchronous invasion of host cells by Toxoplasma gondii.
136
311
34. LericheMA
1991
Characterization of the protein contents of rhoptries and dense granules of Toxoplasma gondii tachyzoites by subcellular fractionation and monoclonal antibodies.
45
249
259
35. Garcia-ReguetN
FourmauxMN
Mercereau-PuijalonO
MannT
The microneme protein MIC3 of Toxoplasma gondii is a secretory adhesin that binds to both the surface of the host cells and the surface of the parasite.
2
353
364
36. MeissnerM
ReissM
ViebigN
TourselC
A family of transmembrane microneme proteins of Toxoplasma gondii contain EGF-like domains and function as escorters.
115
563
574
37. CouvreurG
SadakA
FortierB
1988
Surface antigens of Toxoplasma gondii.
Parasitology
97
Pt 1
10
38. LambrosC
VanderbergJP
1979
Synchronization of Plasmodium falciparum erythrocytic stages in culture.
65
418
420
39. TrangDT
HuyNT
KariuT
TajimaK
KameiK
One-step concentration of malarial parasite-infected red blood cells and removal of contaminating white blood cells.
Malar J
3
Zvýšte si kvalifikáciu online z pohodlia domova
Nemáte účet? Registrujte sa
Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.
