{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,18]],"date-time":"2026-10-18T13:56:20Z","timestamp":1792331780525,"version":"4.4.0"},"reference-count":42,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2023,12,20]],"date-time":"2023-12-20T00:00:00Z","timestamp":1703030400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2023,12,20]],"date-time":"2023-12-20T00:00:00Z","timestamp":1703030400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"crossref","award":["217202\/Z\/19\/Z, FC001076"],"award-info":[{"award-number":["217202\/Z\/19\/Z, FC001076"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"crossref","award":["FC001999"],"award-info":[{"award-number":["FC001999"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100000289","name":"Cancer Research UK","doi-asserted-by":"crossref","award":["FC001076"],"award-info":[{"award-number":["FC001076"]}],"id":[{"id":"10.13039\/501100000289","id-type":"DOI","asserted-by":"crossref"}]},{"name":"UKRI | Medical Research Council","award":["FC001076"],"award-info":[{"award-number":["FC001076"]}]},{"DOI":"10.13039\/501100000289","name":"Cancer Research UK","doi-asserted-by":"crossref","award":["FC001999"],"award-info":[{"award-number":["FC001999"]}],"id":[{"id":"10.13039\/501100000289","id-type":"DOI","asserted-by":"crossref"}]},{"name":"UKRI | Medical Research Council","award":["FC001999"],"award-info":[{"award-number":["FC001999"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mSphere"],"published-print":{"date-parts":[[2023,12,20]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Infection with the parasite\n            <jats:italic>Toxoplasma gondii<\/jats:italic>\n            leads to production of interferon gamma (IFN\u03b3) 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\n            <jats:italic>Toxoplasma<\/jats:italic>\n            , 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\n            <jats:italic>Toxoplasma<\/jats:italic>\n            infection while recruited to the PV in IFN\u03b3-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 IFN\u03b3-dependent control pathway of\n            <jats:italic>Toxoplasma<\/jats:italic>\n            in endothelial cells. We assessed p97\/VCP in\n            <jats:italic>Toxoplasma<\/jats:italic>\n            control in various human cells and demonstrate that p97\/VCP is a universal IFN\u03b3-dependent host restriction factor targeting the\n            <jats:italic>Toxoplasma<\/jats:italic>\n            PV in epithelial (HeLa) and endothelial cells (human umbilical vein endothelial cells), fibroblasts (human foreskin fibroblast), and macrophages (THP1).\n          <\/jats:p>\n          <jats:sec>\n            <jats:title>IMPORTANCE<\/jats:title>\n            <jats:p>\n              <jats:italic>Toxoplasma gondii<\/jats:italic>\n              (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 (IFN\u03b3), 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 IFN\u03b3-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.\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/msphere.00511-23","type":"journal-article","created":{"date-parts":[[2023,11,17]],"date-time":"2023-11-17T14:01:39Z","timestamp":1700229699000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":8,"title":["p97\/VCP targets\n            <i>Toxoplasma gondii<\/i>\n            vacuoles for parasite restriction in interferon-stimulated human cells"],"prefix":"10.1128","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3235-6170","authenticated-orcid":false,"given":"Barbara","family":"Clough","sequence":"first","affiliation":[{"name":"Institute for Microbiology and Infection, School of Biosciences, The University of Birmingham, Birmingham, United Kingdom"},{"name":"Host-Toxoplasma Interaction Laboratory, The Francis Crick Institute, London, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Fisch","sequence":"additional","affiliation":[{"name":"Institute for Microbiology and Infection, School of Biosciences, The University of Birmingham, Birmingham, United Kingdom"},{"name":"Host-Toxoplasma Interaction Laboratory, The Francis Crick Institute, London, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Todd H.","family":"Mize","sequence":"additional","affiliation":[{"name":"Advanced Mass Spectrometry Facility, School of Biosciences, The University of Birmingham, Birmingham, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vesela","family":"Encheva","sequence":"additional","affiliation":[{"name":"Proteomics Science Technology Platform, The Francis Crick Institute, London, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ambrosius","family":"Snijders","sequence":"additional","affiliation":[{"name":"Proteomics Science Technology Platform, The Francis Crick Institute, London, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9515-3442","authenticated-orcid":true,"given":"Eva-Maria","family":"Frickel","sequence":"additional","affiliation":[{"name":"Institute for Microbiology and Infection, School of Biosciences, The University of Birmingham, Birmingham, United Kingdom"},{"name":"Host-Toxoplasma Interaction Laboratory, The Francis Crick Institute, London, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijpara.2009.04.003"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.136.5.1157"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.pt.2017.02.007"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.3128869"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-5822.2010.01443.x"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2017.10.021"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20201314"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006027"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01157-15"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.15252\/embj.2019103758"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00054-20"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.15252\/embj.2018100926"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2020.108008"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.adg2253"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.01888-22"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.81.3.908"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/0166-6851(86)90101-5"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1111\/cmi.12579"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1093\/femspd\/ftab058"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1038\/embor.2008.93"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.E11-09-0817"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M115.710707"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1042\/BCJ20160783"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1111\/cmi.13349"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.15252\/embj.201695148"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006176"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2009.01.010"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.0020034"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1110\/ps.03554604"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1111\/febs.15226"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1003809"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41418-021-00906-9"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1038\/emboj.2009.18"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.093831"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2015.08.028"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncb2301"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-30963-z"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2009.03.012"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-59745-255-7_3"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2009.36"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkab1038"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","unstructured":"Clough B Fisch D Mize TH Encheva V Snijders A Frickel EM. 2023. P97\/VCP targets Toxoplasma Gondii Vacuoles for parasite restriction in interferon-stimulated cells. bioRxiv. doi:10.1101\/2023.06.20.545566:2023.06.20.545566.","DOI":"10.1101\/2023.06.20.545566:2023.06.20.545566"}],"container-title":["mSphere"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/msphere.00511-23","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/msphere.00511-23","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,12,20]],"date-time":"2023-12-20T14:06:56Z","timestamp":1703081216000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/msphere.00511-23"}},"subtitle":[],"editor":[{"given":"Ira J.","family":"Blader","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2023,12,20]]},"references-count":42,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2023,12,20]]}},"alternative-id":["10.1128\/msphere.00511-23"],"URL":"https:\/\/doi.org\/10.1128\/msphere.00511-23","relation":{},"ISSN":["2379-5042"],"issn-type":[{"value":"2379-5042","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,12,20]]},"assertion":[{"value":"2023-09-07","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-10-09","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-11-17","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e00511-23"}}