{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,17]],"date-time":"2025-11-17T21:40:19Z","timestamp":1763415619275,"version":"build-2065373602"},"reference-count":48,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2022,10,8]],"date-time":"2022-10-08T00:00:00Z","timestamp":1665187200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"the German Research Foundation (DFG)","doi-asserted-by":"publisher","award":["STA357\/7-1","STA357\/8-1"],"award-info":[{"award-number":["STA357\/7-1","STA357\/8-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>PML nuclear bodies (PML-NBs) are dynamic macromolecular complexes that mediate intrinsic immunity against viruses of different families, including human cytomegalovirus (HCMV). Upon HCMV infection, PML-NBs target viral genomes entering the nucleus and restrict viral immediate\u2013early gene expression by epigenetic silencing. Studies from several groups performed in human fibroblast cells have shown that the major PML-NB components PML, Daxx, Sp100 and ATRX contribute to this repression in a cooperative manner. Their role for HCMV restriction in endothelial cells, however, has not yet been characterized although infected endothelium is thought to play a crucial role for HCMV dissemination and development of vascular disease in vivo. Here, we use conditionally immortalized umbilical vein endothelial cells (HEC-LTT) as a cell culture model to elucidate the impact of PML-NB proteins on lytic HCMV infection. Depletion of individual PML-NB proteins by lentiviral transduction showed a particularly strong antiviral effect of PML in HEC-LTT, compared to human fibroblasts. A closer characterization of this antiviral function revealed that PML may not only effectively inhibit HCMV immediate-early gene expression but also act at later steps of the viral replication cycle. At contrast, we surprisingly noted an antiviral behavior of Daxx in complementary approaches: Depletion of Daxx resulted in decreased viral gene expression, while overexpression of Daxx promoted HCMV infection. In summary, our data demonstrate a cell type-specific effect of PML-NB components on lytic HCMV infection and suggest an important role of PML in the inhibition of HCMV dissemination through infected endothelial cells.<\/jats:p>","DOI":"10.3390\/ijms231911931","type":"journal-article","created":{"date-parts":[[2022,10,8]],"date-time":"2022-10-08T23:39:31Z","timestamp":1665272371000},"page":"11931","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Promyelocytic Leukemia Protein Potently Restricts Human Cytomegalovirus Infection in Endothelial Cells"],"prefix":"10.3390","volume":"23","author":[{"given":"Sven","family":"Seitz","sequence":"first","affiliation":[{"name":"Institute of Virology, Ulm University Medical Center, 89081 Ulm, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna Theresa","family":"Heusel","sequence":"additional","affiliation":[{"name":"Institute of Virology, Ulm University Medical Center, 89081 Ulm, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9878-3119","authenticated-orcid":false,"given":"Thomas","family":"Stamminger","sequence":"additional","affiliation":[{"name":"Institute of Virology, Ulm University Medical Center, 89081 Ulm, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Myriam","family":"Scherer","sequence":"additional","affiliation":[{"name":"Institute of Virology, Ulm University Medical Center, 89081 Ulm, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1038\/s41579-021-00582-z","article-title":"Pathogenesis of human cytomegalovirus in the immunocompromised host","volume":"19","author":"Griffiths","year":"2021","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1007\/s00404-017-4380-2","article-title":"Cytomegalovirus infection in pregnancy","volume":"296","author":"Kagan","year":"2017","journal-title":"Arch. Gynecol. Obstet."},{"key":"ref_3","first-page":"63","article-title":"Cytomegalovirus cell tropism","volume":"325","author":"Sinzger","year":"2008","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1016\/j.humimm.2004.02.009","article-title":"Pathogenesis of human cytomegalovirus infection and cellular targets","volume":"65","author":"Gerna","year":"2004","journal-title":"Hum. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1057","DOI":"10.1160\/TH09-04-0213","article-title":"Endothelial cells in human cytomegalovirus infection: One host cell out of many or a crucial target for virus spread?","volume":"102","author":"Adler","year":"2009","journal-title":"Thromb. Haemost."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"130","DOI":"10.3389\/fcimb.2020.00130","article-title":"Cytomegalovirus Latency and Reactivation: An Intricate Interplay with the Host Immune Response","volume":"10","author":"Forte","year":"2020","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1093\/abbs\/gmaa005","article-title":"The role of human cytomegalovirus in atherosclerosis: A systematic review","volume":"52","author":"Zhu","year":"2020","journal-title":"Acta Biochim. Biophys. Sin."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"956","DOI":"10.1093\/infdis\/150.6.956","article-title":"Replication of human cytomegalovirus in endothelial cells","volume":"150","author":"Ho","year":"1984","journal-title":"J. Infect. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"127","DOI":"10.2144\/000114326","article-title":"A permanently growing human endothelial cell line supports productive infection with human cytomegalovirus under conditional cell growth arrest","volume":"59","author":"Lieber","year":"2015","journal-title":"BioTechniques"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1089\/ten.tea.2009.0184","article-title":"Synthetic gene regulation circuits for control of cell expansion","volume":"16","author":"May","year":"2010","journal-title":"Tissue Eng. Part A"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1109","DOI":"10.1038\/ni1125","article-title":"Intrinsic immunity: A front-line defense against viral attack","volume":"5","author":"Bieniasz","year":"2004","journal-title":"Nat. Immunol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1128\/JVI.01685-07","article-title":"Nuclear domain 10 components promyelocytic leukemia protein and hDaxx independently contribute to an intrinsic antiviral defense against human cytomegalovirus infection","volume":"82","author":"Tavalai","year":"2008","journal-title":"J. Virol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1532","DOI":"10.1099\/vir.0.030981-0","article-title":"Human cytomegalovirus immediate-early gene expression is restricted by the nuclear domain 10 component Sp100","volume":"92","author":"Adler","year":"2011","journal-title":"J. Gen. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"12543","DOI":"10.1128\/JVI.01215-08","article-title":"Human cytomegalovirus protein pp71 displaces the chromatin-associated factor ATRX from nuclear domain 10 at early stages of infection","volume":"82","author":"Lukashchuk","year":"2008","journal-title":"J. Virol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"8621","DOI":"10.1128\/JVI.00621-16","article-title":"MORC3, a Component of PML Nuclear Bodies, Has a Role in Restricting Herpes Simplex virus 1 and Human Cytomegalovirus","volume":"90","author":"Sloan","year":"2016","journal-title":"J. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1006","DOI":"10.1038\/nrm2277","article-title":"Structure, dynamics and functions of promyelocytic leukaemia nuclear bodies","volume":"8","author":"Bernardi","year":"2007","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1016\/j.ceb.2018.03.011","article-title":"PML nuclear bodies: From architecture to function","volume":"52","year":"2018","journal-title":"Curr. Opin. Cell Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1240","DOI":"10.3390\/v1031240","article-title":"Interplay between Herpesvirus Infection and Host Defense by PML Nuclear Bodies","volume":"1","author":"Tavalai","year":"2009","journal-title":"Viruses"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e73006","DOI":"10.7554\/eLife.73006","article-title":"Dual signaling via interferon and DNA damage response elicits entrapment by giant PML nuclear bodies","volume":"11","author":"Scherer","year":"2022","journal-title":"eLife"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3863","DOI":"10.1128\/JVI.80.8.3863-3871.2006","article-title":"Inactivating a cellular intrinsic immune defense mediated by Daxx is the mechanism through which the human cytomegalovirus pp71 protein stimulates viral immediate-early gene expression","volume":"80","author":"Saffert","year":"2006","journal-title":"J. Virol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"6527","DOI":"10.1128\/JVI.78.12.6527-6542.2004","article-title":"Ability of the human cytomegalovirus IE1 protein to modulate sumoylation of PML correlates with its functional activities in transcriptional regulation and infectivity in cultured fibroblast cells","volume":"78","author":"Lee","year":"2004","journal-title":"J. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e02049-16","DOI":"10.1128\/JVI.02049-16","article-title":"The Human Cytomegalovirus IE1 Protein Antagonizes PML Nuclear Body-Mediated Intrinsic Immunity via the Inhibition of PML De Novo SUMOylation","volume":"91","author":"Schilling","year":"2017","journal-title":"J. Virol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"9447","DOI":"10.1128\/JVI.00870-11","article-title":"Evidence for a dual antiviral role of the major nuclear domain 10 component Sp100 during the immediate-early and late phases of the human cytomegalovirus replication cycle","volume":"85","author":"Tavalai","year":"2011","journal-title":"J. Virol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"10635","DOI":"10.1073\/pnas.1937626100","article-title":"The ATRX syndrome protein forms a chromatin-remodeling complex with Daxx and localizes in promyelocytic leukemia nuclear bodies","volume":"100","author":"Xue","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1006\/excr.1996.0353","article-title":"The nuclear domain 10 (ND10) is disrupted by the human cytomegalovirus gene product IE1","volume":"229","author":"Korioth","year":"1996","journal-title":"Exp. Cell Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"7995","DOI":"10.1128\/JVI.00734-06","article-title":"PML contributes to a cellular mechanism of repression of herpes simplex virus type 1 infection that is inactivated by ICP0","volume":"80","author":"Everett","year":"2006","journal-title":"J. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1099\/vir.0.83286-0","article-title":"Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40\/E","volume":"89","author":"Sinzger","year":"2008","journal-title":"J. Gen. Virol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"8006","DOI":"10.1128\/JVI.00743-06","article-title":"Evidence for a role of the cellular ND10 protein PML in mediating intrinsic immunity against human cytomegalovirus infections","volume":"80","author":"Tavalai","year":"2006","journal-title":"J. Virol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1089\/jir.2010.0111","article-title":"Role of promyelocytic leukemia protein in host antiviral defense","volume":"31","author":"Geoffroy","year":"2011","journal-title":"J. Interferon Cytokine Res. Off. J. Int. Soc. Interferon Cytokine Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1128\/JVI.02023-12","article-title":"The adenoviral oncogene E1A-13S interacts with a specific isoform of the tumor suppressor PML to enhance viral transcription","volume":"87","author":"Berscheminski","year":"2013","journal-title":"J. Virol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4581","DOI":"10.1242\/jcs.112.24.4581","article-title":"Cell cycle regulation of PML modification and ND10 composition","volume":"112","author":"Everett","year":"1999","journal-title":"J. Cell Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"37652","DOI":"10.1074\/jbc.M604273200","article-title":"Human Daxx-mediated repression of human cytomegalovirus gene expression correlates with a repressive chromatin structure around the major immediate early promoter","volume":"281","author":"Woodhall","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1083\/jcb.134.4.815","article-title":"The periphery of nuclear domain 10 (ND10) as site of DNA virus deposition","volume":"134","author":"Ishov","year":"1996","journal-title":"J. Cell Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1083\/jcb.138.1.5","article-title":"Human cytomegalovirus immediate early interaction with host nuclear structures: Definition of an immediate transcript environment","volume":"138","author":"Ishov","year":"1997","journal-title":"J. Cell Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"5078","DOI":"10.1128\/JVI.79.8.5078-5089.2005","article-title":"ND10 components relocate to sites associated with herpes simplex virus type 1 nucleoprotein complexes during virus infection","volume":"79","author":"Everett","year":"2005","journal-title":"J. Virol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2174","DOI":"10.1128\/JVI.02950-12","article-title":"Components of promyelocytic leukemia nuclear bodies (ND10) act cooperatively to repress herpesvirus infection","volume":"87","author":"Glass","year":"2013","journal-title":"J. Virol."},{"key":"ref_37","first-page":"261","article-title":"Daxx, a broad-spectrum viral restriction factor","volume":"20","author":"Maillet","year":"2016","journal-title":"Virologie"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2442","DOI":"10.1038\/s41467-022-30134-9","article-title":"Identification of DAXX as a restriction factor of SARS-CoV-2 through a CRISPR\/Cas9 screen","volume":"13","author":"Maarifi","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1186\/s12985-015-0335-z","article-title":"DAXX modulates human papillomavirus early gene expression and genome replication in U2OS cells","volume":"12","author":"Ustav","year":"2015","journal-title":"Virol. J."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1038\/s41586-021-03824-5","article-title":"DAXX represents a new type of protein-folding enabler","volume":"597","author":"Huang","year":"2021","journal-title":"Nature"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2884","DOI":"10.3390\/v7062751","article-title":"Contribution of the Major ND10 Proteins PML, hDaxx and Sp100 to the Regulation of Human Cytomegalovirus Latency and Lytic Replication in the Monocytic Cell Line THP-1","volume":"7","author":"Wagenknecht","year":"2015","journal-title":"Viruses"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Stilp, A.C., Scherer, M., K\u00f6nig, P., F\u00fcrstberger, A., Kestler, H.A., and Stamminger, T. (2022). The chromatin remodeling protein ATRX positively regulates IRF3-dependent type I interferon production and interferon-induced gene expression. PLoS Pathog., 18.","DOI":"10.1371\/journal.ppat.1010748"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/0166-0934(89)90129-8","article-title":"A rapid microneutralization assay for the measurement of neutralizing antibody reactive with human cytomegalovirus","volume":"23","author":"Andreoni","year":"1989","journal-title":"J. Virol. Methods"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2510","DOI":"10.1128\/JVI.74.6.2510-2524.2000","article-title":"Covalent modification of the transactivator protein IE2-p86 of human cytomegalovirus by conjugation to the ubiquitin-homologous proteins SUMO-1 and hSMT3b","volume":"74","author":"Hofmann","year":"2000","journal-title":"J. Virol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"7048","DOI":"10.1128\/jvi.71.9.7048-7060.1997","article-title":"The UL84 protein of human cytomegalovirus acts as a transdominant inhibitor of immediate-early-mediated transactivation that is able to prevent viral replication","volume":"71","author":"Gebert","year":"1997","journal-title":"J. Virol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/0168-1702(92)90123-Q","article-title":"The DNA-binding protein P52 of human cytomegalovirus reacts with monoclonal antibody CCH2 and associates with the nuclear membrane at late times after infection","volume":"24","author":"Plachter","year":"1992","journal-title":"Virus Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"3943","DOI":"10.1128\/jvi.68.6.3943-3954.1994","article-title":"UL69 of human cytomegalovirus, an open reading frame with homology to ICP27 of herpes simplex virus, encodes a transactivator of gene expression","volume":"68","author":"Winkler","year":"1994","journal-title":"J. Virol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1016\/S0140-6736(89)91144-6","article-title":"Non-specific mesangial staining with antibodies against cytomegalovirus in immunoglobulin-A nephropathy","volume":"1","author":"Waldo","year":"1989","journal-title":"Lancet"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/19\/11931\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:47:59Z","timestamp":1760143679000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/19\/11931"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,8]]},"references-count":48,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2022,10]]}},"alternative-id":["ijms231911931"],"URL":"https:\/\/doi.org\/10.3390\/ijms231911931","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2022,10,8]]}}}