A heterotrimeric complex of Toxoplasma proteins promotes parasite survival in interferon gamma-stimulated human cells.

Toxoplasma gondii secretes protein effectors to subvert the human immune system sufficiently to establish a chronic infection. Relative to murine infections, little is known about which parasite effectors disarm human immune responses. Here, we used targeted CRISPR screening to identify secreted pro...

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Main Authors: Eloise J Lockyer, Francesca Torelli, Simon Butterworth, Ok-Ryul Song, Steven Howell, Anne Weston, Philip East, Moritz Treeck
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2023-07-01
Series:PLoS Biology
Online Access:https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.3002202&type=printable
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author Eloise J Lockyer
Francesca Torelli
Simon Butterworth
Ok-Ryul Song
Steven Howell
Anne Weston
Philip East
Moritz Treeck
author_facet Eloise J Lockyer
Francesca Torelli
Simon Butterworth
Ok-Ryul Song
Steven Howell
Anne Weston
Philip East
Moritz Treeck
author_sort Eloise J Lockyer
collection DOAJ
description Toxoplasma gondii secretes protein effectors to subvert the human immune system sufficiently to establish a chronic infection. Relative to murine infections, little is known about which parasite effectors disarm human immune responses. Here, we used targeted CRISPR screening to identify secreted protein effectors required for parasite survival in IFNγ-activated human cells. Independent screens were carried out using 2 Toxoplasma strains that differ in virulence in mice, leading to the identification of effectors required for survival in IFNγ-activated human cells. We identify the secreted protein GRA57 and 2 other proteins, GRA70 and GRA71, that together form a complex which enhances the ability of parasites to persist in IFNγ-activated human foreskin fibroblasts (HFFs). Components of the protein machinery required for export of Toxoplasma proteins into the host cell were also found to be important for parasite resistance to IFNγ in human cells, but these export components function independently of the identified protein complex. Host-mediated ubiquitination of the parasite vacuole has previously been associated with increased parasite clearance from human cells, but we find that vacuoles from GRA57, GRA70, and GRA71 knockout strains are surprisingly less ubiquitinated by the host cell. We hypothesise that this is likely a secondary consequence of deletion of the complex, unlinked to the IFNγ resistance mediated by these effectors.
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institution Kabale University
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publisher Public Library of Science (PLoS)
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spelling doaj-art-d87d73d3d44044a985b2ccb89a6e12f52025-08-20T03:25:20ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852023-07-01217e300220210.1371/journal.pbio.3002202A heterotrimeric complex of Toxoplasma proteins promotes parasite survival in interferon gamma-stimulated human cells.Eloise J LockyerFrancesca TorelliSimon ButterworthOk-Ryul SongSteven HowellAnne WestonPhilip EastMoritz TreeckToxoplasma gondii secretes protein effectors to subvert the human immune system sufficiently to establish a chronic infection. Relative to murine infections, little is known about which parasite effectors disarm human immune responses. Here, we used targeted CRISPR screening to identify secreted protein effectors required for parasite survival in IFNγ-activated human cells. Independent screens were carried out using 2 Toxoplasma strains that differ in virulence in mice, leading to the identification of effectors required for survival in IFNγ-activated human cells. We identify the secreted protein GRA57 and 2 other proteins, GRA70 and GRA71, that together form a complex which enhances the ability of parasites to persist in IFNγ-activated human foreskin fibroblasts (HFFs). Components of the protein machinery required for export of Toxoplasma proteins into the host cell were also found to be important for parasite resistance to IFNγ in human cells, but these export components function independently of the identified protein complex. Host-mediated ubiquitination of the parasite vacuole has previously been associated with increased parasite clearance from human cells, but we find that vacuoles from GRA57, GRA70, and GRA71 knockout strains are surprisingly less ubiquitinated by the host cell. We hypothesise that this is likely a secondary consequence of deletion of the complex, unlinked to the IFNγ resistance mediated by these effectors.https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.3002202&type=printable
spellingShingle Eloise J Lockyer
Francesca Torelli
Simon Butterworth
Ok-Ryul Song
Steven Howell
Anne Weston
Philip East
Moritz Treeck
A heterotrimeric complex of Toxoplasma proteins promotes parasite survival in interferon gamma-stimulated human cells.
PLoS Biology
title A heterotrimeric complex of Toxoplasma proteins promotes parasite survival in interferon gamma-stimulated human cells.
title_full A heterotrimeric complex of Toxoplasma proteins promotes parasite survival in interferon gamma-stimulated human cells.
title_fullStr A heterotrimeric complex of Toxoplasma proteins promotes parasite survival in interferon gamma-stimulated human cells.
title_full_unstemmed A heterotrimeric complex of Toxoplasma proteins promotes parasite survival in interferon gamma-stimulated human cells.
title_short A heterotrimeric complex of Toxoplasma proteins promotes parasite survival in interferon gamma-stimulated human cells.
title_sort heterotrimeric complex of toxoplasma proteins promotes parasite survival in interferon gamma stimulated human cells
url https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.3002202&type=printable
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