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An apical membrane complex controls rhoptry exocytosis and invasion in Toxoplasma

Sparvoli, Daniela and Delabre, Jason and Penarete-Vargas, Diana Marcela and Mageswaran, Shrawan Kumar and Tsypin, Lev M. and Heckendorn, Justine and Theveny, Liam and Maynadier, Marjorie and Cova, Marta Mendonça and Berry-Sterkers, Laurence and Guérin, Amandine and Dubremetz, Jean-François and Urbach, Serge and Striepen, Boris and Turkewitz, Aaron P. and Chang, Yi-Wei and Lebrun, Maryse (2022) An apical membrane complex controls rhoptry exocytosis and invasion in Toxoplasma. . (Unpublished) https://resolver.caltech.edu/CaltechAUTHORS:20220228-619488000

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Abstract

Apicomplexan parasites possess secretory organelles called rhoptries that undergo regulated exocytosis upon contact with the host. This process is essential for the parasitic lifestyle of these pathogens and relies on an exocytic machinery sharing structural features and molecular components with free-living ciliates. Here, we performed a Tetrahymena-based transcriptomic screen to uncover novel exocytic factors in Ciliata and Apicomplexa. We identified membrane-bound proteins, named CRMPs, forming part of a large complex essential for rhoptry secretion and invasion in Toxoplasma. In contrast to previously described rhoptry exocytic factors, TgCRMPs are not required for the assembly of the rhoptry secretion machinery and only transiently associated with the exocytic site - prior to invasion. CRMPs and their partners contain putative host cell-binding domains, and CRMPa shares similarity to GPCR proteins. We propose that the CRMP complex acts as host-molecular sensor to ensure that rhoptry exocytosis occurs when the parasite contacts the host cell.


Item Type:Report or Paper (Discussion Paper)
Related URLs:
URLURL TypeDescription
https://doi.org/10.1101/2022.02.25.481937DOIDiscussion Paper
ORCID:
AuthorORCID
Sparvoli, Daniela0000-0002-5992-3753
Penarete-Vargas, Diana Marcela0000-0002-3306-3943
Mageswaran, Shrawan Kumar0000-0002-0803-7129
Tsypin, Lev M.0000-0002-0642-8468
Theveny, Liam0000-0002-6292-8408
Cova, Marta Mendonça0000-0001-6727-248X
Berry-Sterkers, Laurence0000-0002-8150-7860
Guérin, Amandine0000-0001-8307-5099
Dubremetz, Jean-François0000-0002-1362-9110
Urbach, Serge0000-0001-8663-2006
Striepen, Boris0000-0002-7426-432X
Turkewitz, Aaron P.0000-0003-3531-5806
Chang, Yi-Wei0000-0003-2391-473X
Lebrun, Maryse0000-0002-0962-0156
Additional Information:The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. We thank Sebastian Lourido for the pU6-Universal plasmid, Dominique Soldati-Favre for providing the anti-ARM (ARO) antibodies and pLinker-2xTy-DHFR plasmid, Nicolas Dos Santos Pacheco for helping in setting up the Ultrastructure Expansion Microscopy, Anita Koshy for the toxofilin-Cre plasmid and Helen Blau’s lab for the Cre reporter DSred cell line. We thank Veronique Richard and Frank Godiard of the MEA platform, University of Montpellier for their assistance with electron microscopy and Pilar Ruga Fahy of the Pôle Facultaire de Microscopie Ultrastructurale, in Geneva for preparation of freeze-fracture replicas. We are also grateful to Elodie Jublanc, Vicky Diakou and the imaging facility MRI at the University of Montpellier, part of the national infrastructure France-BioImaging supported by the French National Research Agency (ANR-10-INBS-04, «Investments for the future»), and Christophe Duperray of the MRI-Cytometry at the Institute for Regenerative Medicine and Biotherapy for their assistance and technical support. Mass spectrometry experiments were carried out using the facilities of the Montpellier Proteomics Platform (PPM, BioCampus Montpellier). We thank Stefan Steimle for his technical assistance with the Krios G3i cryogenic electron microscope; the Singh Center for Nanotechnology and the Beckman Center for Cryogenic Electron Microscopy at the University of Pennsylvania for hosting and supporting the use of the Titan Krios. Dr Maryse Lebrun is an INSERM researcher. This work was supported by the Laboratoire d’Excellence (LabEx) (ParaFrap ANR-11-LABX-0024), and European Research Council (ERC advanced grant number 833309 KissAndSpitRhoptry) to M.L.; by the FACCTS (France and Chicago Collaborating in the Sciences) to A.P.T and M.L. ; by NIH GM105783 to A.P.T. ; by a David and Lucile Packard Fellowship for Science and Engineering (2019-69645) and a Pennsylvania Department of Health FY19 Health Research Formula Fund to Y.-W.C; by NIH R01 AI112427 to B. S. D.S. and M.M.C. are supported by the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program under Grant agreement no. 833309 to M.L. AUTHOR CONTRIBUTION. Conceptualization: D.S. M.L. Investigation Tetrahymena: L.M.T. performed CDH analysis; D.S. generated and analyzed mutants. Investigation Toxoplasma: J.D., J.H., D.M.P.V. generated the tagged and iKD lines; J.D. performed phenotypic analysis of the mutants with the help of M.M., D.M.P.V., J.H.; D.S. generated and analyzed lines for co-IP and fluorescence microscopy of extracellular parasites; D.M.P.V., S.U., performed IP and mass spectrometry analysis; L.B.S., J.H. performed EM analysis; D.S., M.M.C., performed ultrastructure expansion microscopy. Phylogenies: D.S. Freeze fracture data: D.S., J.H. prepared samples; D.S., J.F.D., collected data. Cryo-ET data: A.G. cultured the cells; A.G. and S.K.M. prepared the grids; S.K.M. collected data; S.K.M. and L.T. performed analysis on the tomograms and prepared figures with guidance from Y.-W.C.; L.T. performed subtomogram averaging for the CRMPb-iKD mutant with input from Y.-W.C. and S.K.M. Writing-Original draft: D.S., M.L. Visualization: D.S., J.D., M.L. Writing-Review & Editing: A.P.T., B.S., J.D., S.K.M, with inputs from all authors. Supervision: A.P.T., Y.-W.C., M.L. Funding: B.S., A.P.T., Y.-W.C., M.L. DATA AVAILABILITY. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE (Perez-Riverol et al., 2019) partner repository with the dataset identifier PXD031161 and PXD031164. The authors have declared no competing interest.
Funders:
Funding AgencyGrant Number
Agence Nationale pour la Recherche (ANR)ANR-10-INBS-04
Agence Nationale pour la Recherche (ANR)ANR-11-LABX-0024
European Research Council (ERC)833309
France And Chicago Collaborating in The Sciences (FACCTS)UNSPECIFIED
NIHGM105783
David and Lucile Packard Foundation2019-69645
Pennsylvania Department of HealthUNSPECIFIED
NIHR01 AI112427
Subject Keywords:Apicomplexa, Ciliates, Plasmodium, Toxoplasma, Rhoptry, exocytosis, invasion, CRMP, Alveolata, signaling
DOI:10.1101/2022.02.25.481937
Record Number:CaltechAUTHORS:20220228-619488000
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20220228-619488000
Official Citation:An apical membrane complex controls rhoptry exocytosis and invasion in Toxoplasma Daniela Sparvoli, Jason Delabre, Diana Marcela Penarete-Vargas, Shrawan Kumar Mageswaran, Lev M. Tsypin, Justine Heckendorn, Liam Theveny, Marjorie Maynadier, Marta Mendonça Cova, Laurence Berry-Sterkers, Amandine Guérin, Jean-François Dubremetz, Serge Urbach, Boris Striepen, Aaron P. Turkewitz, Yi-Wei Chang, Maryse Lebrun bioRxiv 2022.02.25.481937; doi: https://doi.org/10.1101/2022.02.25.481937
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:113625
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:01 Mar 2022 18:53
Last Modified:01 Mar 2022 18:53

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