Host Cell invasion
The Apicomplexa phylum is defined by the presence of an apical complex of organelles comprising two specific secretory granules called micronemes and rhoptries, which sequentially release their contents during invasion. Microneme proteins contribute to motility, attachment and invasion, while rhoptry proteins contribute to both invasion and subsequent manipulation of host cell functions after the parasites have established themselves in their host cells. Our research is focused on the formation of a structure (the moving junction) which is essential for host cell invasion in both Toxoplasma and Plasmodium, and which depends on the cooperation between microneme and rhoptry proteins. We are also interested on the mechanism of secretion of rhoptries, which remains a major conundrum in the field.
Schematic of T. gondii tachyzoite invading a host cell. Details in the inset show how, in the MJ, AMA1 is exposed on the parasite surface and interacts with a short extracellular domain of RON2 incorporated into the membrane of the host cell. The complex of RON2, RON4, RON5 and RON8 localize to the host cytosol and cooperate to recruit adaptor proteins (ALIX, CD2AP, CIN85, TSG101) through multiple specific interaction motifs that physically link the RONs complex to the cortical actin cytoskeleton. This results in the stabilization of the junction between the parasite and the host cortical actin providing a substantial cellular anchor for the AMA1/RON2 bridge at the surface, which is necessary to sustain the parasite invasive force.
Rhoptry secretion
The ability of Apicomplexa to cause disease depends on the coordinated secretion of specialized secretory organelles. The rhoptries are particularly important, because they act as the apicomplexan equivalent of bacterial secretion systems. They inject parasite proteins directly in the cytoplasm of host cells not only for invasion but also to hijack host functions crucial to establish and maintain infection. However, in contrast to bacteria where the secretion machinery has been resolved in details at the atomic scale, how these eukaryotic parasites secrete and inject rhoptry effectors into cells is still an enigma. We aim to dissect the mechanistic steps and the molecular components that assemble the rhoptry secretion machine.
The group has been been awarded ERC Advanced grant 2018 over 5 years by the European Council to unravel the mechanism of eukaryote secretion system.
An Alveolata secretory machinery adapted to parasite host cell invasion.
Aquilini E, Cova MM, Mageswaran SK, Dos Santos Pacheco N, Sparvoli D, Penarete-Vargas DM, Najm R, Graindorge A, Suarez C, Maynadier M, Berry-Sterkers L, Urbach S, Fahy PR, Guérin AN, Striepen B, Dubremetz JF, Chang YW, Turkewitz AP, Lebrun M.Nat Microbiol. 2021 Jan 25. doi: 10.1038/s41564-020-00854-z. Online ahead of print.PMID: 33495622
Co-option of Plasmodium falciparum PP1 for egress from host erythrocytes.
Paul AS, Miliu A, Paulo JA, Goldberg JM, Bonilla AM, Berry L, Seveno M, Braun-Breton C, Kosber AL, Elsworth B, Arriola JSN, Lebrun M, Gygi SP, Lamarque MH, Duraisingh MT.Nat Commun. 2020 Jul 15;11(1):3532. doi: 10.1038/s41467-020-17306-1.PMID: 32669539
A Toxoplasma gondii patatin-like phospholipase contributes to host cell invasion.
Wilson SK, Heckendorn J, Martorelli Di Genova B, Koch LL, Rooney PJ, Morrissette N, Lebrun M, Knoll LJ.PLoS Pathog. 2020 Jul 6;16(7):e1008650. doi: 10.1371/journal.ppat.1008650. eCollection 2020 Jul.PMID: 32628723
Assessing Rhoptry Secretion in T. gondii.
Suarez C, Lodoen MB, Lebrun M.Methods Mol Biol. 2020;2071:143-155. doi: 10.1007/978-1-4939-9857-9_8.PMID: 31758451
TgZFP2 is a novel zinc finger protein involved in coordinating mitosis and budding in Toxoplasma.
Semenovskaya K, Lévêque MF, Berry L, Bordat Y, Dubremetz JF, Lebrun M, Besteiro S.Cell Microbiol. 2020 Jan;22(1):e13120. doi: 10.1111/cmi.13120. Epub 2019 Nov 13.PMID: 31628778
A lipid-binding protein mediates rhoptry discharge and invasion in Plasmodium falciparum and Toxoplasma gondii parasites.
Suarez C, Lentini G, Ramaswamy R, Maynadier M, Aquilini E, Berry-Sterkers L, Cipriano M, Chen AL, Bradley P, Striepen B, Boulanger MJ, Lebrun M.Nat Commun. 2019 Sep 6;10(1):4041. doi: 10.1038/s41467-019-11979-z.PMID: 31492901
Berry L, Chen CT, Francia ME, Guerin A, Graindorge A, Saliou JM, Grandmougin M, Wein S, Bechara C, Morlon-Guyot J, Bordat Y, Gubbels MJ, Lebrun M, Dubremetz JF, Daher W. Toxoplasma gondii chromosomal passenger complex is essential for the organization of a functional mitotic spindle: a prerequisite for productive endodyogeny. Cell Mol Life Sci, (2018) Jul 26.
Morlon-Guyot J, El Hajj H, Martin K, Fois A, Carrillo A, Berry L, Burchmore R, Meissner M, Lebrun M, Daher W. A proteomic analysis unravels novel CORVET and HOPS proteins involved in Toxoplasma gondii secretory organelles biogenesis. Cell Microbiol, (2018) 17:e12870.
Wengelnik K, Daher W, Lebrun M. Phosphoinositides and their functions in apicomplexan parasites. Int J Parasitol. (2018) Jun;48(7):493-504.
Lebrun M, Blanchard N. Host-microbe interactions: parasites. Volume 40 December 2017
Guérin A, El Hajj H, Penarete-Vargas D, Besteiro S, Lebrun M. RON4(L1) is a new member of the moving junction complex in Toxoplasma gondii. Sci Rep, (2017) Dec 20;7(1):17907.
Lentini G, El Hajj H, Papoin J, Fall G, Pfaff AW, Tawil N, Braun-Breton C, Lebrun M. Characterization of Toxoplasma DegP, a rhoptry serine protease crucial for lethal infection in mice. PLoS One, (2017) Dec 15;12(12):e0189556.
Frénal K, Dubremetz JF, Lebrun M, Soldati-Favre D. Gliding motility powers invasion and egress in Apicomplexa. Nat Rev Microbiol, (2017) Nov;15(11):645-660.
Guérin A, Corrales RM, Parker ML, Lamarque MH, Jacot D, El Hajj H, Soldati-Favre D, Boulanger MJ, Lebrun M. Efficient invasion by Toxoplasma depends on the subversion of host protein networks. Nat Microbiol, (2017) Oct;2(10):1358-1366.
Miliu A, Lebrun M, Braun-Breton C, Lamarque MH. Shelph2, a bacterial-like phosphatase of the malaria parasite Plasmodium falciparum, is dispensable during asexual blood stage. PLoS One, (2017) Oct 26;12(10):e0187073.
Nguyen HM, El Hajj H, El Hajj R, Tawil N, Berry L, Lebrun M, Bordat Y, Besteiro S. Toxoplasma gondii autophagy-related protein ATG9 is crucial for the survival of parasites in their host. Cell Microbiol, (2016) Jun (19) 6.
Berry L, Chen CT, Reininger L, Carvalho TG, El Hajj H, Morlon-Guyot J, Bordat Y, Lebrun M, Gubbels MJ, Doerig C, Daher W. The conserved apicomplexan Aurora kinase TgArk3 is involved in endodyogeny, duplication rate and parasite virulence. Cell Microbiol, (2016) Aug;18(8):1106-20.
Daher W, Morlon-Guyot J, Alayi TD, Tomavo S, Wengelnik K, Lebrun M. Identification of Toxoplasma TgPH1, a pleckstrin homology domain-containing protein that binds to the phosphoinositide PI(3,5)P2. Mol Biochem Parasitol, (2016) May;207(1):39-44.
Parker ML, Penarete-Vargas DM, Hamilton PT, Guérin A, Dubey JP, Perlman SJ, Spano F, Lebrun M*, Boulanger MJ*. Dissecting the interface between apicomplexan parasite and host cell: Insights from a divergent AMA-RON2 pair. PNAS, (2016) Jan 12;113(2):398-403. *equal contribution
Wang K, Peng ED, Huang AS, Xia D, Vermont SJ, Lentini G, Lebrun M, Wastling JM, Bradley PJ. Identification of Novel O-Linked Glycosylated Toxoplasma Proteins by Vicia villosa Lectin Chromatography. PLoS One, (2016) Mar 7;11(3):e0150561.
Delgadillo RF, Parker ML, Lebrun M, Boulanger MJ, Douguet D. Stability of the Plasmodium falciparum AMA1-RON2 Complex Is Governed by the Domain II (DII) Loop. PLoS One, (2016) Jan 5;11(1):e0144764.
Pihan E., Delgadillo RF., Tonkin ML., Pugnière M., Lebrun M., Boulanger MJ., Douguet D. Computational and biophysical approaches to protein-protein interaction inhibition of Plasmodium falciparum AMA1/RON2 complex. J Comput Aided Mol Des, (2015) Jun;29(6):525-39
Rugarabamu G., Marq JB., Guérin A., Lebrun M., Soldati-Favre D. Distinct contribution of Toxoplasma gondii rhomboid proteases 4 and 5 to micronemal protein protease 1 activity during invasion. Mol Microbiol, (2015). Jul;97(2):244-62.
Lentini G., Kong-Hap M., El Hajj H., Francia M., Claudet C., Striepen B., Dubremetz JF., Lebrun M. Identification and characterization of Toxoplasma SIP, a conserved apicomplexan cytoskeleton protein involved in maintaining the shape, motility and virulence of the parasite. Cell Microbiol, (2015) 17(1):62-78.
Morlon-Guyot J., Pastore S., Berry L., Lebrun M., Daher W. Toxoplasma gondii Vps11, a subunit of HOPS and CORVET tethering complexes, is essential for the biogenesis of secretory organelles. Cell Microbiol, (2015) 1157-1178.
Daher W., Morlon-Guyot J., Sheiner L., Lentini G., Berry L., Tawk L., Dubremetz JF., Wengelnik K., Striepen B., Lebrun M. Lipid kinases are essential for apicoplast homeostasis in Toxoplasma gondii. Cell Microbiol, (2015) 559-578.
Risco-Castillo V., Topçu Selma., Marinach C., Manzon G., Bigorgne A.E., Briquet S., Baudin X., Lebrun M., Dubremetz J.F., and Olivier Silvie. Malaria Sporozoites Traverse Host Cells within Transient Vacuoles. Cell Host & Microbes, (2015) Nov 11;18(5):593-603.
Lamarque MH., Roques M., Kong-Hap M., Tonkin ML., Rugarabamu G., Marq JB., Penarete-Vargas DM., Boulanger MJ., Soldati-Favre D., Lebrun M. Plasticity and redundancy among AMA-RON pairs ensure host cell entry of Toxoplasma parasites. Nat Commun, (2014) Jun 17;5:4098.
Morlon-Guyot J., Berry L., Chen CT., Gubbels MJ., Lebrun M., Daher W. The Toxoplasma gondii calcium-dependent protein kinase 7 is involved in early steps of parasite division and is crucial for parasite survival. Cell Microbiol, (2014) 95-114.
Kong-Hap MA., Mouammine A., Daher W., Berry L., Lebrun M., Dubremetz JF., Besteiro S. Regulation of ATG8 membrane association by ATG4 in the parasitic protist Toxoplasma gondii. Autophagy, (2013) September ;9(9):1334-48
Tonkin ML., Crawford J., Lebrun ML., Boulanger MJ. Babesia divergens and Neospora caninum apical membrane antigen 1 structures reveal selectivity and plasticity in apicomplexan parasite host cell invasion. Protein Sci, (2011) Jan;22(1):114-27
De Napoli. M.G., N. de Miguel., Lebrun M., Moreno S.N.J., Angel S.O., Corvi M.M. N-terminal palmitoylation is required for Toxoplasma gondii HSP20 inner membrane complex localization. Biochimica and Biophysica Acta (BBA) - Molecular Cell Research, (2013) Jun;1833(6):1329-37.
Lebrun, M., Carruthers, V.B., Cesbron-Delauw, M.-F. Toxoplasma Secretory Proteins and their roles in Cell Invasion and Intracellular Survival. in Toxoplasma gondii: The Model Apicomplexan – Perspectives and Methods, Weiss and Kim (Eds), 2nd edition (2013) 265-315.
Dubremetz JF, Lebrun M. Virulence factors of Toxoplasma gondii. Microbes Infect, (2012) Dec;14(15):1403-10.
Lamarque MH, Papoin J, Finizio AL, Lentini G, Pfaff AW, Candolfi E, Dubremetz JF, Lebrun M. Identification of a new rhoptry neck complex RON9/RON10 in the Apicomplexa parasite Toxoplasma gondii. PLoS One, (2012);7(3):e32457.
Vulliez-Le Normand B, Tonkin ML, Lamarque MH, Langer S, Hoos S, Roques M, Saul FA, Faber BW, Bentley GA, Boulanger MJ, Lebrun M. Structural and functional insights into the malaria parasite moving junction complex. PLoS Pathog, (2012);8(6):e1002755.
Tawk, L., Dubremetz, J. F., Montcourrier, P., Chicanne, G., Merezegue, F., Richard, V., Payrastre, B., Meissner, M., Vial, H. J., Roy, C., Wengelnik, K., and Lebrun, M. Phosphatidylinositol 3-monophosphate is involved in Toxoplasma apicoplast biogenesis. PLoS Pathog, (2011) 7, e1001286
Besteiro S, Dubremetz JF, Lebrun M. The moving junction of apicomplexan parasites: a key structure for invasion. Cell Microbiol, (2011) Jun;13(6):797-805.
Lamarque M, Besteiro S, Papoin J, Roques M, Vulliez-Le Normand B, Morlon-Guyot J, Dubremetz JF, Fauquenoy S, Tomavo S, Faber BW, Kocken CH, Thomas AW, Boulanger MJ, Bentley GA, Lebrun M. The RON2-AMA1 interaction is a critical step in moving junction-dependent invasion by apicomplexan parasites. PLoS Pathog, (2011) Feb 10;7(2):e1001276.
Hajagos BE, Turetzky JM, Peng ED, Cheng SJ, Ryan CM, Souda P, Whitelegge JP, Lebrun M, Dubremetz JF, Bradley PJ. Molecular dissection of novel trafficking and processing of the Toxoplasma gondii rhoptry metalloprotease toxolysin-1. Traffic, (2012) Feb;13(2):292-304.
Tonkin ML, Roques M, Lamarque MH, Pugnière M, Douguet D, Crawford J, Lebrun M*, Boulanger MJ*. Host cell invasion by apicomplexan parasites: insights from the co-structure of AMA1 with a RON2 peptide. Science, (2011) Jul 22;333(6041):463-7. *equal contribution
