Translocation of Dense Granule Effectors across the Parasitophorous Vacuole Membrane in <italic toggle="yes">Toxoplasma-</italic>Infected Cells Requires the Activity of ROP17, a Rhoptry Protein Kinase

ABSTRACT Toxoplasma gondii tachyzoites co-opt host cell functions through introduction of a large set of rhoptry- and dense granule-derived effector proteins. These effectors reach the host cytosol through different means: direct injection for rhoptry effectors and translocation across the parasitop...

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Autores principales:	Michael W. Panas, Abel Ferrel, Adit Naor, Elizabeth Tenborg, Hernan A. Lorenzi, John C. Boothroyd
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Toxoplasma gondii effector proteins host-parasite relationship kinases Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/9d8fdf4a8eaa474880523088ef5e0042
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spelling	oai:doaj.org-article:9d8fdf4a8eaa474880523088ef5e00422021-11-15T15:22:27ZTranslocation of Dense Granule Effectors across the Parasitophorous Vacuole Membrane in <italic toggle="yes">Toxoplasma-</italic>Infected Cells Requires the Activity of ROP17, a Rhoptry Protein Kinase10.1128/mSphere.00276-192379-5042https://doaj.org/article/9d8fdf4a8eaa474880523088ef5e00422019-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00276-19https://doaj.org/toc/2379-5042ABSTRACT Toxoplasma gondii tachyzoites co-opt host cell functions through introduction of a large set of rhoptry- and dense granule-derived effector proteins. These effectors reach the host cytosol through different means: direct injection for rhoptry effectors and translocation across the parasitophorous vacuolar membrane (PVM) for dense granule (GRA) effectors. The machinery that translocates these GRA effectors has recently been partially elucidated, revealing three components, MYR1, MYR2, and MYR3. To determine whether other proteins might be involved, we returned to a library of mutants defective in GRA translocation and selected one with a partial defect, suggesting it might be in a gene encoding a new component of the machinery. Surprisingly, whole-genome sequencing revealed a missense mutation in a gene encoding a known rhoptry protein, a serine/threonine protein kinase known as ROP17. ROP17 resides on the host cytosol side of the PVM in infected cells and has previously been known for its activity in phosphorylating and thereby inactivating host immunity-related GTPases. Here, we show that null or catalytically dead mutants of ROP17 are defective in GRA translocation across the PVM but that translocation can be rescued “in trans” by ROP17 delivered by other tachyzoites infecting the same host cell. This strongly argues that ROP17’s role in regulating GRA translocation is carried out on the host cytosolic side of the PVM, not within the parasites or lumen of the parasitophorous vacuole. This represents an entirely new way in which the different secretory compartments of Toxoplasma tachyzoites collaborate to modulate the host-parasite interaction. IMPORTANCE When Toxoplasma infects a cell, it establishes a protective parasitophorous vacuole surrounding it. While this vacuole provides protection, it also serves as a barrier to the export of parasite effector proteins that impact and take control of the host cell. Our discovery here that the parasite rhoptry protein ROP17 is necessary for export of these effector proteins provides a distinct, novel function for ROP17 apart from its known role in protecting the vacuole. This will enable future research into ways in which we can prevent the export of effector proteins, thereby preventing Toxoplasma from productively infecting its animal and human hosts.Michael W. PanasAbel FerrelAdit NaorElizabeth TenborgHernan A. LorenziJohn C. BoothroydAmerican Society for MicrobiologyarticleToxoplasma gondiieffector proteinshost-parasite relationshipkinasesMicrobiologyQR1-502ENmSphere, Vol 4, Iss 4 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Toxoplasma gondii effector proteins host-parasite relationship kinases Microbiology QR1-502
spellingShingle	Toxoplasma gondii effector proteins host-parasite relationship kinases Microbiology QR1-502 Michael W. Panas Abel Ferrel Adit Naor Elizabeth Tenborg Hernan A. Lorenzi John C. Boothroyd Translocation of Dense Granule Effectors across the Parasitophorous Vacuole Membrane in <italic toggle="yes">Toxoplasma-</italic>Infected Cells Requires the Activity of ROP17, a Rhoptry Protein Kinase
description	ABSTRACT Toxoplasma gondii tachyzoites co-opt host cell functions through introduction of a large set of rhoptry- and dense granule-derived effector proteins. These effectors reach the host cytosol through different means: direct injection for rhoptry effectors and translocation across the parasitophorous vacuolar membrane (PVM) for dense granule (GRA) effectors. The machinery that translocates these GRA effectors has recently been partially elucidated, revealing three components, MYR1, MYR2, and MYR3. To determine whether other proteins might be involved, we returned to a library of mutants defective in GRA translocation and selected one with a partial defect, suggesting it might be in a gene encoding a new component of the machinery. Surprisingly, whole-genome sequencing revealed a missense mutation in a gene encoding a known rhoptry protein, a serine/threonine protein kinase known as ROP17. ROP17 resides on the host cytosol side of the PVM in infected cells and has previously been known for its activity in phosphorylating and thereby inactivating host immunity-related GTPases. Here, we show that null or catalytically dead mutants of ROP17 are defective in GRA translocation across the PVM but that translocation can be rescued “in trans” by ROP17 delivered by other tachyzoites infecting the same host cell. This strongly argues that ROP17’s role in regulating GRA translocation is carried out on the host cytosolic side of the PVM, not within the parasites or lumen of the parasitophorous vacuole. This represents an entirely new way in which the different secretory compartments of Toxoplasma tachyzoites collaborate to modulate the host-parasite interaction. IMPORTANCE When Toxoplasma infects a cell, it establishes a protective parasitophorous vacuole surrounding it. While this vacuole provides protection, it also serves as a barrier to the export of parasite effector proteins that impact and take control of the host cell. Our discovery here that the parasite rhoptry protein ROP17 is necessary for export of these effector proteins provides a distinct, novel function for ROP17 apart from its known role in protecting the vacuole. This will enable future research into ways in which we can prevent the export of effector proteins, thereby preventing Toxoplasma from productively infecting its animal and human hosts.
format	article
author	Michael W. Panas Abel Ferrel Adit Naor Elizabeth Tenborg Hernan A. Lorenzi John C. Boothroyd
author_facet	Michael W. Panas Abel Ferrel Adit Naor Elizabeth Tenborg Hernan A. Lorenzi John C. Boothroyd
author_sort	Michael W. Panas
title	Translocation of Dense Granule Effectors across the Parasitophorous Vacuole Membrane in <italic toggle="yes">Toxoplasma-</italic>Infected Cells Requires the Activity of ROP17, a Rhoptry Protein Kinase
title_short	Translocation of Dense Granule Effectors across the Parasitophorous Vacuole Membrane in <italic toggle="yes">Toxoplasma-</italic>Infected Cells Requires the Activity of ROP17, a Rhoptry Protein Kinase
title_full	Translocation of Dense Granule Effectors across the Parasitophorous Vacuole Membrane in <italic toggle="yes">Toxoplasma-</italic>Infected Cells Requires the Activity of ROP17, a Rhoptry Protein Kinase
title_fullStr	Translocation of Dense Granule Effectors across the Parasitophorous Vacuole Membrane in <italic toggle="yes">Toxoplasma-</italic>Infected Cells Requires the Activity of ROP17, a Rhoptry Protein Kinase
title_full_unstemmed	Translocation of Dense Granule Effectors across the Parasitophorous Vacuole Membrane in <italic toggle="yes">Toxoplasma-</italic>Infected Cells Requires the Activity of ROP17, a Rhoptry Protein Kinase
title_sort	translocation of dense granule effectors across the parasitophorous vacuole membrane in <italic toggle="yes">toxoplasma-</italic>infected cells requires the activity of rop17, a rhoptry protein kinase
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/9d8fdf4a8eaa474880523088ef5e0042
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Translocation of Dense Granule Effectors across the Parasitophorous Vacuole Membrane in <italic toggle="yes">Toxoplasma-</italic>Infected Cells Requires the Activity of ROP17, a Rhoptry Protein Kinase

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