The Secreted Acid Phosphatase Domain-Containing GRA44 from <named-content content-type="genus-species">Toxoplasma gondii</named-content> Is Required for c-Myc Induction in Infected Cells

ABSTRACT During host cell invasion, the eukaryotic pathogen Toxoplasma gondii forms a parasitophorous vacuole to safely reside within the cell, while it is partitioned from host cell defense mechanisms. From within this safe niche, parasites sabotage multiple host cell systems, including gene expres...

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Autores principales:	William J. Blakely, Michael J. Holmes, Gustavo Arrizabalaga
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	GRA44 MYR1 Toxoplasma c-Myc phosphatase translocation Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/0ae5a106e4684bbc8fd846b7dd46a6c6
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spelling	oai:doaj.org-article:0ae5a106e4684bbc8fd846b7dd46a6c62021-11-15T15:27:53ZThe Secreted Acid Phosphatase Domain-Containing GRA44 from <named-content content-type="genus-species">Toxoplasma gondii</named-content> Is Required for c-Myc Induction in Infected Cells10.1128/mSphere.00877-192379-5042https://doaj.org/article/0ae5a106e4684bbc8fd846b7dd46a6c62020-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00877-19https://doaj.org/toc/2379-5042ABSTRACT During host cell invasion, the eukaryotic pathogen Toxoplasma gondii forms a parasitophorous vacuole to safely reside within the cell, while it is partitioned from host cell defense mechanisms. From within this safe niche, parasites sabotage multiple host cell systems, including gene expression, apoptosis, and intracellular immune recognition, by secreting a large arsenal of effector proteins. Many parasite proteins studied for active host cell manipulative interactions have been kinases. The translocation of effectors from the parasitophorous vacuole into the host cell is mediated by a putative translocon complex, which includes the proteins MYR1, MYR2, and MYR3. Whether other proteins are involved in the structure or regulation of this putative translocon is not known. We have discovered that the secreted protein GRA44, which contains a putative acid phosphatase domain, interacts with members of this complex and is required for host cell effects downstream of effector secretion. We have determined that GRA44 is processed in a region with homology to sequences targeted by protozoan proteases of the secretory pathway and that both major cleavage fragments are secreted into the parasitophorous vacuole. Immunoprecipitation experiments showed that GRA44 interacts with a large number of secreted proteins, including MYR1. Importantly, conditional knockdown of GRA44 resulted in a lack of host cell c-Myc upregulation, which mimics the phenotype seen when members of the translocon complex are genetically disrupted. Thus, the putative acid phosphatase GRA44 is crucial for host cell alterations during Toxoplasma infection and is associated with the translocon complex which Toxoplasma relies upon for success as an intracellular pathogen. IMPORTANCE Approximately one-third of humans are infected with the parasite Toxoplasma gondii. Toxoplasma infections can lead to severe disease in those with a compromised or suppressed immune system. Additionally, infections during pregnancy present a significant health risk to the developing fetus. Drugs that target this parasite are limited, have significant side effects, and do not target all disease stages. Thus, a thorough understanding of how the parasite propagates within a host is critical in the discovery of novel therapeutic targets. Toxoplasma replication requires that it enter the cells of the infected organism. In order to survive the environment inside a cell, Toxoplasma secretes a large repertoire of proteins, which hijack a number of important cellular functions. How these Toxoplasma proteins move from the parasite into the host cell is not well understood. Our work shows that the putative phosphatase GRA44 is part of a protein complex responsible for this process.William J. BlakelyMichael J. HolmesGustavo ArrizabalagaAmerican Society for MicrobiologyarticleGRA44MYR1Toxoplasmac-MycphosphatasetranslocationMicrobiologyQR1-502ENmSphere, Vol 5, Iss 1 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	GRA44 MYR1 Toxoplasma c-Myc phosphatase translocation Microbiology QR1-502
spellingShingle	GRA44 MYR1 Toxoplasma c-Myc phosphatase translocation Microbiology QR1-502 William J. Blakely Michael J. Holmes Gustavo Arrizabalaga The Secreted Acid Phosphatase Domain-Containing GRA44 from <named-content content-type="genus-species">Toxoplasma gondii</named-content> Is Required for c-Myc Induction in Infected Cells
description	ABSTRACT During host cell invasion, the eukaryotic pathogen Toxoplasma gondii forms a parasitophorous vacuole to safely reside within the cell, while it is partitioned from host cell defense mechanisms. From within this safe niche, parasites sabotage multiple host cell systems, including gene expression, apoptosis, and intracellular immune recognition, by secreting a large arsenal of effector proteins. Many parasite proteins studied for active host cell manipulative interactions have been kinases. The translocation of effectors from the parasitophorous vacuole into the host cell is mediated by a putative translocon complex, which includes the proteins MYR1, MYR2, and MYR3. Whether other proteins are involved in the structure or regulation of this putative translocon is not known. We have discovered that the secreted protein GRA44, which contains a putative acid phosphatase domain, interacts with members of this complex and is required for host cell effects downstream of effector secretion. We have determined that GRA44 is processed in a region with homology to sequences targeted by protozoan proteases of the secretory pathway and that both major cleavage fragments are secreted into the parasitophorous vacuole. Immunoprecipitation experiments showed that GRA44 interacts with a large number of secreted proteins, including MYR1. Importantly, conditional knockdown of GRA44 resulted in a lack of host cell c-Myc upregulation, which mimics the phenotype seen when members of the translocon complex are genetically disrupted. Thus, the putative acid phosphatase GRA44 is crucial for host cell alterations during Toxoplasma infection and is associated with the translocon complex which Toxoplasma relies upon for success as an intracellular pathogen. IMPORTANCE Approximately one-third of humans are infected with the parasite Toxoplasma gondii. Toxoplasma infections can lead to severe disease in those with a compromised or suppressed immune system. Additionally, infections during pregnancy present a significant health risk to the developing fetus. Drugs that target this parasite are limited, have significant side effects, and do not target all disease stages. Thus, a thorough understanding of how the parasite propagates within a host is critical in the discovery of novel therapeutic targets. Toxoplasma replication requires that it enter the cells of the infected organism. In order to survive the environment inside a cell, Toxoplasma secretes a large repertoire of proteins, which hijack a number of important cellular functions. How these Toxoplasma proteins move from the parasite into the host cell is not well understood. Our work shows that the putative phosphatase GRA44 is part of a protein complex responsible for this process.
format	article
author	William J. Blakely Michael J. Holmes Gustavo Arrizabalaga
author_facet	William J. Blakely Michael J. Holmes Gustavo Arrizabalaga
author_sort	William J. Blakely
title	The Secreted Acid Phosphatase Domain-Containing GRA44 from <named-content content-type="genus-species">Toxoplasma gondii</named-content> Is Required for c-Myc Induction in Infected Cells
title_short	The Secreted Acid Phosphatase Domain-Containing GRA44 from <named-content content-type="genus-species">Toxoplasma gondii</named-content> Is Required for c-Myc Induction in Infected Cells
title_full	The Secreted Acid Phosphatase Domain-Containing GRA44 from <named-content content-type="genus-species">Toxoplasma gondii</named-content> Is Required for c-Myc Induction in Infected Cells
title_fullStr	The Secreted Acid Phosphatase Domain-Containing GRA44 from <named-content content-type="genus-species">Toxoplasma gondii</named-content> Is Required for c-Myc Induction in Infected Cells
title_full_unstemmed	The Secreted Acid Phosphatase Domain-Containing GRA44 from <named-content content-type="genus-species">Toxoplasma gondii</named-content> Is Required for c-Myc Induction in Infected Cells
title_sort	secreted acid phosphatase domain-containing gra44 from <named-content content-type="genus-species">toxoplasma gondii</named-content> is required for c-myc induction in infected cells
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/0ae5a106e4684bbc8fd846b7dd46a6c6
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The Secreted Acid Phosphatase Domain-Containing GRA44 from <named-content content-type="genus-species">Toxoplasma gondii</named-content> Is Required for c-Myc Induction in Infected Cells

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