p97/VCP targets Toxoplasma gondii vacuoles for parasite restriction in interferon-stimulated human cells
ABSTRACTInfection with the parasite Toxoplasma gondii leads to production of interferon gamma (IFNγ) that stimulates cells to upregulate defense proteins targeting the parasite for cell intrinsic elimination or growth restriction. Various host defense mechanisms operate at the parasitophorous vacuol...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2023-12-01
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| Series: | mSphere |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/msphere.00511-23 |
| _version_ | 1797384684097765376 |
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| author | Barbara Clough Daniel Fisch Todd H. Mize Vesela Encheva Ambrosius Snijders Eva-Maria Frickel |
| author_facet | Barbara Clough Daniel Fisch Todd H. Mize Vesela Encheva Ambrosius Snijders Eva-Maria Frickel |
| author_sort | Barbara Clough |
| collection | DOAJ |
| description | ABSTRACTInfection with the parasite Toxoplasma gondii leads to production of interferon gamma (IFNγ) that stimulates cells to upregulate defense proteins targeting the parasite for cell intrinsic elimination or growth restriction. Various host defense mechanisms operate at the parasitophorous vacuole (PV) in different human cell types leading to PV disruption, acidification, or membrane envelopment. Ubiquitin and p62 are players in all human host control mechanisms of Toxoplasma, but other unifying proteins have not been identified. Here, we show that p97/valosin-containing protein (VCP), as well as its associated proteins ANKRD13A and UBXD1 control Toxoplasma infection while recruited to the PV in IFNγ-stimulated endothelial cells. Convergent deposition of ANKRD13A, p97/VCP, and UBXD1 onto the same vacuole is dependent on vacuolar ubiquitination and observed within 2 h post-infection. ANKRD13A, p97/VCP, and UBXD1 all drive the acidification mechanism of the vacuole, which is the IFNγ-dependent control pathway of Toxoplasma in endothelial cells. We assessed p97/VCP in Toxoplasma control in various human cells and demonstrate that p97/VCP is a universal IFNγ-dependent host restriction factor targeting the Toxoplasma PV in epithelial (HeLa) and endothelial cells (human umbilical vein endothelial cells), fibroblasts (human foreskin fibroblast), and macrophages (THP1).IMPORTANCEToxoplasma gondii (Tg) is a ubiquitous parasitic pathogen, infecting about one-third of the global population. Tg is controlled in immunocompetent people by mechanisms that are not fully understood. Tg infection drives the production of the inflammatory cytokine interferon gamma (IFNγ), which upregulates intracellular anti-pathogen defense pathways. In this study, we describe host proteins p97/VCP, UBXD1, and ANKRD13A that control Tg at the parasitophorous vacuole (PV) in IFNγ-stimulated endothelial cells. p97/VCP is an ATPase that interacts with a network of cofactors and is active in a wide range of ubiquitin-dependent cellular processes. We demonstrate that PV ubiquitination is a pre-requisite for recruitment of these host defense proteins, and their deposition directs Tg PVs to acidification in endothelial cells. We show that p97/VCP universally targets PVs in human cells and restricts Tg in different human cell types. Overall, these findings reveal new players of intracellular host defense of a vacuolated pathogen. |
| first_indexed | 2024-03-08T21:40:14Z |
| format | Article |
| id | doaj.art-895407d7e0a14877a13c4bbd2aa74765 |
| institution | Directory Open Access Journal |
| issn | 2379-5042 |
| language | English |
| last_indexed | 2024-03-08T21:40:14Z |
| publishDate | 2023-12-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mSphere |
| spelling | doaj.art-895407d7e0a14877a13c4bbd2aa747652023-12-20T14:01:03ZengAmerican Society for MicrobiologymSphere2379-50422023-12-018610.1128/msphere.00511-23p97/VCP targets Toxoplasma gondii vacuoles for parasite restriction in interferon-stimulated human cellsBarbara Clough0Daniel Fisch1Todd H. Mize2Vesela Encheva3Ambrosius Snijders4Eva-Maria Frickel5Institute for Microbiology and Infection, School of Biosciences, The University of Birmingham, Birmingham, United KingdomInstitute for Microbiology and Infection, School of Biosciences, The University of Birmingham, Birmingham, United KingdomAdvanced Mass Spectrometry Facility, School of Biosciences, The University of Birmingham, Birmingham, United KingdomProteomics Science Technology Platform, The Francis Crick Institute, London, United KingdomProteomics Science Technology Platform, The Francis Crick Institute, London, United KingdomInstitute for Microbiology and Infection, School of Biosciences, The University of Birmingham, Birmingham, United KingdomABSTRACTInfection with the parasite Toxoplasma gondii leads to production of interferon gamma (IFNγ) that stimulates cells to upregulate defense proteins targeting the parasite for cell intrinsic elimination or growth restriction. Various host defense mechanisms operate at the parasitophorous vacuole (PV) in different human cell types leading to PV disruption, acidification, or membrane envelopment. Ubiquitin and p62 are players in all human host control mechanisms of Toxoplasma, but other unifying proteins have not been identified. Here, we show that p97/valosin-containing protein (VCP), as well as its associated proteins ANKRD13A and UBXD1 control Toxoplasma infection while recruited to the PV in IFNγ-stimulated endothelial cells. Convergent deposition of ANKRD13A, p97/VCP, and UBXD1 onto the same vacuole is dependent on vacuolar ubiquitination and observed within 2 h post-infection. ANKRD13A, p97/VCP, and UBXD1 all drive the acidification mechanism of the vacuole, which is the IFNγ-dependent control pathway of Toxoplasma in endothelial cells. We assessed p97/VCP in Toxoplasma control in various human cells and demonstrate that p97/VCP is a universal IFNγ-dependent host restriction factor targeting the Toxoplasma PV in epithelial (HeLa) and endothelial cells (human umbilical vein endothelial cells), fibroblasts (human foreskin fibroblast), and macrophages (THP1).IMPORTANCEToxoplasma gondii (Tg) is a ubiquitous parasitic pathogen, infecting about one-third of the global population. Tg is controlled in immunocompetent people by mechanisms that are not fully understood. Tg infection drives the production of the inflammatory cytokine interferon gamma (IFNγ), which upregulates intracellular anti-pathogen defense pathways. In this study, we describe host proteins p97/VCP, UBXD1, and ANKRD13A that control Tg at the parasitophorous vacuole (PV) in IFNγ-stimulated endothelial cells. p97/VCP is an ATPase that interacts with a network of cofactors and is active in a wide range of ubiquitin-dependent cellular processes. We demonstrate that PV ubiquitination is a pre-requisite for recruitment of these host defense proteins, and their deposition directs Tg PVs to acidification in endothelial cells. We show that p97/VCP universally targets PVs in human cells and restricts Tg in different human cell types. Overall, these findings reveal new players of intracellular host defense of a vacuolated pathogen.https://journals.asm.org/doi/10.1128/msphere.00511-23Toxoplasma gondiiinnate immunityparasitophorous vacuoleubiquitinANKRD13Ap97/VCP |
| spellingShingle | Barbara Clough Daniel Fisch Todd H. Mize Vesela Encheva Ambrosius Snijders Eva-Maria Frickel p97/VCP targets Toxoplasma gondii vacuoles for parasite restriction in interferon-stimulated human cells mSphere Toxoplasma gondii innate immunity parasitophorous vacuole ubiquitin ANKRD13A p97/VCP |
| title | p97/VCP targets Toxoplasma gondii vacuoles for parasite restriction in interferon-stimulated human cells |
| title_full | p97/VCP targets Toxoplasma gondii vacuoles for parasite restriction in interferon-stimulated human cells |
| title_fullStr | p97/VCP targets Toxoplasma gondii vacuoles for parasite restriction in interferon-stimulated human cells |
| title_full_unstemmed | p97/VCP targets Toxoplasma gondii vacuoles for parasite restriction in interferon-stimulated human cells |
| title_short | p97/VCP targets Toxoplasma gondii vacuoles for parasite restriction in interferon-stimulated human cells |
| title_sort | p97 vcp targets toxoplasma gondii vacuoles for parasite restriction in interferon stimulated human cells |
| topic | Toxoplasma gondii innate immunity parasitophorous vacuole ubiquitin ANKRD13A p97/VCP |
| url | https://journals.asm.org/doi/10.1128/msphere.00511-23 |
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