ISG15 Connects Autophagy and IFN-γ-Dependent Control of <named-content content-type="genus-species">Toxoplasma gondii</named-content> Infection in Human Cells

ABSTRACT The intracellular protozoan parasite Toxoplasma gondii is capable of infecting most nucleated cells, where it survives in a specially modified compartment called the parasitophorous vacuole (PV). Interferon gamma (IFN-γ) is the major cytokine involved in activating cell-autonomous immune re...

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Autores principales:	Jaya Bhushan, Joshua B. Radke, Yi-Chieh Perng, Michael Mcallaster, Deborah J. Lenschow, Herbert W. Virgin, L. David Sibley
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	ATG5 BioID ubiquitin autophagy adaptors LC3 ISGylation Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/076f57152f6140a48b7e5dad96971507
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spelling	oai:doaj.org-article:076f57152f6140a48b7e5dad969715072021-11-15T16:19:09ZISG15 Connects Autophagy and IFN-γ-Dependent Control of <named-content content-type="genus-species">Toxoplasma gondii</named-content> Infection in Human Cells10.1128/mBio.00852-202150-7511https://doaj.org/article/076f57152f6140a48b7e5dad969715072020-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00852-20https://doaj.org/toc/2150-7511ABSTRACT The intracellular protozoan parasite Toxoplasma gondii is capable of infecting most nucleated cells, where it survives in a specially modified compartment called the parasitophorous vacuole (PV). Interferon gamma (IFN-γ) is the major cytokine involved in activating cell-autonomous immune responses to inhibit parasite growth within this intracellular niche. In HeLa cells, IFN-γ treatment leads to ubiquitination of susceptible parasite strains, recruitment of the adaptors p62 and NDP52, and engulfment in microtubule-associated protein 1 light chain 3 (LC3)-positive membranes that restrict parasite growth. IFN-γ-mediated growth restriction depends on core members of the autophagy (ATG) pathway but not the initiation or degradative steps in the process. To explore the connection between these different pathways, we used permissive biotin ligation to identify proteins that interact with ATG5 in an IFN-γ-dependent fashion. Network analysis of the ATG5 interactome identified interferon-stimulated gene 15 (ISG15), which is highly upregulated by IFN treatment, as a hub connecting the ATG complex with other IFN-γ-induced genes, suggesting that it forms a functional link between the pathways. Deletion of ISG15 resulted in impaired recruitment of p62, NDP52, and LC3 to the PV and loss of IFN-γ-restricted parasite growth. The function of ISG15 required conjugation, and a number of ISGylated targets overlapped with the IFN-γ-dependent ATG5 interactome, including the adapter p62. Collectively, our findings establish a role for ISG15 in connecting the ATG pathway with IFN-γ-dependent restriction of T. gondii in human cells. IMPORTANCE Interferon(s) provide the primary defense against intracellular pathogens, a property ascribed to their ability to upregulate interferon-stimulated genes. Due to the sequestered niche occupied by Toxoplasma gondii, the host has elaborated intricate ways to target the parasite within its vacuole. One such mechanism is the recognition by a noncanonical autophagy pathway that envelops the parasite-containing vacuole and stunts growth in human cells. Remarkably, autophagy-dependent growth restriction requires interferon-γ, yet none of the classical components of autophagy are induced by interferon. Our studies draw a connection between these pathways by demonstrating that the antiviral protein ISG15, which is normally upregulated by interferons, links the autophagy-mediated control to ubiquitination of the vacuole. These findings suggest a similar link between interferon-γ signaling and autophagy that may underlie defense against other intracellular pathogens.Jaya BhushanJoshua B. RadkeYi-Chieh PerngMichael McallasterDeborah J. LenschowHerbert W. VirginL. David SibleyAmerican Society for MicrobiologyarticleATG5BioIDubiquitinautophagy adaptorsLC3ISGylationMicrobiologyQR1-502ENmBio, Vol 11, Iss 5 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	ATG5 BioID ubiquitin autophagy adaptors LC3 ISGylation Microbiology QR1-502
spellingShingle	ATG5 BioID ubiquitin autophagy adaptors LC3 ISGylation Microbiology QR1-502 Jaya Bhushan Joshua B. Radke Yi-Chieh Perng Michael Mcallaster Deborah J. Lenschow Herbert W. Virgin L. David Sibley ISG15 Connects Autophagy and IFN-γ-Dependent Control of <named-content content-type="genus-species">Toxoplasma gondii</named-content> Infection in Human Cells
description	ABSTRACT The intracellular protozoan parasite Toxoplasma gondii is capable of infecting most nucleated cells, where it survives in a specially modified compartment called the parasitophorous vacuole (PV). Interferon gamma (IFN-γ) is the major cytokine involved in activating cell-autonomous immune responses to inhibit parasite growth within this intracellular niche. In HeLa cells, IFN-γ treatment leads to ubiquitination of susceptible parasite strains, recruitment of the adaptors p62 and NDP52, and engulfment in microtubule-associated protein 1 light chain 3 (LC3)-positive membranes that restrict parasite growth. IFN-γ-mediated growth restriction depends on core members of the autophagy (ATG) pathway but not the initiation or degradative steps in the process. To explore the connection between these different pathways, we used permissive biotin ligation to identify proteins that interact with ATG5 in an IFN-γ-dependent fashion. Network analysis of the ATG5 interactome identified interferon-stimulated gene 15 (ISG15), which is highly upregulated by IFN treatment, as a hub connecting the ATG complex with other IFN-γ-induced genes, suggesting that it forms a functional link between the pathways. Deletion of ISG15 resulted in impaired recruitment of p62, NDP52, and LC3 to the PV and loss of IFN-γ-restricted parasite growth. The function of ISG15 required conjugation, and a number of ISGylated targets overlapped with the IFN-γ-dependent ATG5 interactome, including the adapter p62. Collectively, our findings establish a role for ISG15 in connecting the ATG pathway with IFN-γ-dependent restriction of T. gondii in human cells. IMPORTANCE Interferon(s) provide the primary defense against intracellular pathogens, a property ascribed to their ability to upregulate interferon-stimulated genes. Due to the sequestered niche occupied by Toxoplasma gondii, the host has elaborated intricate ways to target the parasite within its vacuole. One such mechanism is the recognition by a noncanonical autophagy pathway that envelops the parasite-containing vacuole and stunts growth in human cells. Remarkably, autophagy-dependent growth restriction requires interferon-γ, yet none of the classical components of autophagy are induced by interferon. Our studies draw a connection between these pathways by demonstrating that the antiviral protein ISG15, which is normally upregulated by interferons, links the autophagy-mediated control to ubiquitination of the vacuole. These findings suggest a similar link between interferon-γ signaling and autophagy that may underlie defense against other intracellular pathogens.
format	article
author	Jaya Bhushan Joshua B. Radke Yi-Chieh Perng Michael Mcallaster Deborah J. Lenschow Herbert W. Virgin L. David Sibley
author_facet	Jaya Bhushan Joshua B. Radke Yi-Chieh Perng Michael Mcallaster Deborah J. Lenschow Herbert W. Virgin L. David Sibley
author_sort	Jaya Bhushan
title	ISG15 Connects Autophagy and IFN-γ-Dependent Control of <named-content content-type="genus-species">Toxoplasma gondii</named-content> Infection in Human Cells
title_short	ISG15 Connects Autophagy and IFN-γ-Dependent Control of <named-content content-type="genus-species">Toxoplasma gondii</named-content> Infection in Human Cells
title_full	ISG15 Connects Autophagy and IFN-γ-Dependent Control of <named-content content-type="genus-species">Toxoplasma gondii</named-content> Infection in Human Cells
title_fullStr	ISG15 Connects Autophagy and IFN-γ-Dependent Control of <named-content content-type="genus-species">Toxoplasma gondii</named-content> Infection in Human Cells
title_full_unstemmed	ISG15 Connects Autophagy and IFN-γ-Dependent Control of <named-content content-type="genus-species">Toxoplasma gondii</named-content> Infection in Human Cells
title_sort	isg15 connects autophagy and ifn-γ-dependent control of <named-content content-type="genus-species">toxoplasma gondii</named-content> infection in human cells
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/076f57152f6140a48b7e5dad96971507
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ISG15 Connects Autophagy and IFN-γ-Dependent Control of <named-content content-type="genus-species">Toxoplasma gondii</named-content> Infection in Human Cells

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