p97/VCP targets Toxoplasma gondii vacuoles for parasite restriction in interferon-stimulated human cells

• Published in: mSphere Vol. 8; no. 6; p. e0051123
• Main Authors: Clough, Barbara, Fisch, Daniel, Mize, Todd H, Encheva, Vesela, Snijders, Ambrosius, Frickel, Eva-Maria
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 20.12.2023
• Subjects: Acidification; Amino acids; Animals; ANKRD13A; Cell culture; Cell death; Defense mechanisms; Endothelial Cells; Fibroblasts; Host-Microbial Interactions; Humans; Infections; Inflammation; innate immunity; Interferon-gamma; Interferons - metabolism; Intracellular; Macrophages; Mass spectrometry; p97/VCP; Parasites; Parasitophorous vacuole; Pathogens; Peptides; Proteins; Research Article; Scientific imaging; Toxoplasma; Toxoplasma - metabolism; Toxoplasma gondii; Ubiquitin; Ubiquitination; Umbilical vein; Vacuoles; Vacuoles - metabolism; Valosin Containing Protein - metabolism; Valosin-containing protein; γ-Interferon; Toxoplasma gondii; innate immunity; UBXD1; p97/VCP; parasitophorous vacuole; ANKRD13A; ubiquitin
• ISSN: 2379-5042; 2379-5042
• DOI: 10.1128/msphere.00511-23

(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.