Differential Impacts on Host Transcription by ROP and GRA Effectors from the Intracellular Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>

ABSTRACT The intracellular parasite Toxoplasma gondii employs a vast array of effector proteins from the rhoptry and dense granule organelles to modulate host cell biology; these effectors are known as ROPs and GRAs, respectively. To examine the individual impacts of ROPs and GRAs on host gene expre...

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Autores principales:	Suchita Rastogi, Yuan Xue, Stephen R. Quake, John C. Boothroyd
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	Toxoplasma effector functions host-parasite relationship parasitology single-cell RNA sequencing Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/03eae627a1cc42e2baa6737e3792e041
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spelling	oai:doaj.org-article:03eae627a1cc42e2baa6737e3792e0412021-11-15T15:56:46ZDifferential Impacts on Host Transcription by ROP and GRA Effectors from the Intracellular Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>10.1128/mBio.00182-202150-7511https://doaj.org/article/03eae627a1cc42e2baa6737e3792e0412020-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00182-20https://doaj.org/toc/2150-7511ABSTRACT The intracellular parasite Toxoplasma gondii employs a vast array of effector proteins from the rhoptry and dense granule organelles to modulate host cell biology; these effectors are known as ROPs and GRAs, respectively. To examine the individual impacts of ROPs and GRAs on host gene expression, we developed a robust, novel protocol to enrich for ultrapure populations of a naturally occurring and reproducible population of host cells called uninfected-injected (U-I) cells, which Toxoplasma injects with ROPs but subsequently fails to invade. We then performed single-cell transcriptomic analysis at 1 to 3 h postinfection on U-I cells (as well as on uninfected and infected controls) arising from infection with either wild-type parasites or parasites lacking the MYR1 protein, which is required for soluble GRAs to cross the parasitophorous vacuole membrane (PVM) and reach the host cell cytosol. Based on comparisons of infected and U-I cells, the host’s earliest response to infection appears to be driven primarily by the injected ROPs, which appear to induce immune and cellular stress pathways. These ROP-dependent proinflammatory signatures appear to be counteracted by at least some of the MYR1-dependent GRAs and may be enhanced by the MYR-independent GRAs (which are found embedded within the PVM). Finally, signatures detected in uninfected bystander cells from the infected monolayers suggest that MYR1-dependent paracrine effects also counteract inflammatory ROP-dependent processes. IMPORTANCE This work performs transcriptomic analysis of U-I cells, captures the earliest stage of a host cell’s interaction with Toxoplasma gondii, and dissects the effects of individual classes of parasite effectors on host cell biology.Suchita RastogiYuan XueStephen R. QuakeJohn C. BoothroydAmerican Society for MicrobiologyarticleToxoplasmaeffector functionshost-parasite relationshipparasitologysingle-cell RNA sequencingMicrobiologyQR1-502ENmBio, Vol 11, Iss 3 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Toxoplasma effector functions host-parasite relationship parasitology single-cell RNA sequencing Microbiology QR1-502
spellingShingle	Toxoplasma effector functions host-parasite relationship parasitology single-cell RNA sequencing Microbiology QR1-502 Suchita Rastogi Yuan Xue Stephen R. Quake John C. Boothroyd Differential Impacts on Host Transcription by ROP and GRA Effectors from the Intracellular Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>
description	ABSTRACT The intracellular parasite Toxoplasma gondii employs a vast array of effector proteins from the rhoptry and dense granule organelles to modulate host cell biology; these effectors are known as ROPs and GRAs, respectively. To examine the individual impacts of ROPs and GRAs on host gene expression, we developed a robust, novel protocol to enrich for ultrapure populations of a naturally occurring and reproducible population of host cells called uninfected-injected (U-I) cells, which Toxoplasma injects with ROPs but subsequently fails to invade. We then performed single-cell transcriptomic analysis at 1 to 3 h postinfection on U-I cells (as well as on uninfected and infected controls) arising from infection with either wild-type parasites or parasites lacking the MYR1 protein, which is required for soluble GRAs to cross the parasitophorous vacuole membrane (PVM) and reach the host cell cytosol. Based on comparisons of infected and U-I cells, the host’s earliest response to infection appears to be driven primarily by the injected ROPs, which appear to induce immune and cellular stress pathways. These ROP-dependent proinflammatory signatures appear to be counteracted by at least some of the MYR1-dependent GRAs and may be enhanced by the MYR-independent GRAs (which are found embedded within the PVM). Finally, signatures detected in uninfected bystander cells from the infected monolayers suggest that MYR1-dependent paracrine effects also counteract inflammatory ROP-dependent processes. IMPORTANCE This work performs transcriptomic analysis of U-I cells, captures the earliest stage of a host cell’s interaction with Toxoplasma gondii, and dissects the effects of individual classes of parasite effectors on host cell biology.
format	article
author	Suchita Rastogi Yuan Xue Stephen R. Quake John C. Boothroyd
author_facet	Suchita Rastogi Yuan Xue Stephen R. Quake John C. Boothroyd
author_sort	Suchita Rastogi
title	Differential Impacts on Host Transcription by ROP and GRA Effectors from the Intracellular Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_short	Differential Impacts on Host Transcription by ROP and GRA Effectors from the Intracellular Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_full	Differential Impacts on Host Transcription by ROP and GRA Effectors from the Intracellular Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_fullStr	Differential Impacts on Host Transcription by ROP and GRA Effectors from the Intracellular Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_full_unstemmed	Differential Impacts on Host Transcription by ROP and GRA Effectors from the Intracellular Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>
title_sort	differential impacts on host transcription by rop and gra effectors from the intracellular parasite <named-content content-type="genus-species">toxoplasma gondii</named-content>
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/03eae627a1cc42e2baa6737e3792e041
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Differential Impacts on Host Transcription by ROP and GRA Effectors from the Intracellular Parasite <named-content content-type="genus-species">Toxoplasma gondii</named-content>

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