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.
abstract: Promyelocytic leukemia nuclear bodies, also termed nuclear domain 10 (ND10), have emerged as nuclear protein accumulations mediating an intrinsic cellular defense against viral infections via chromatin-based mechanisms, however, their contribution to the control of herpesviral latency is still controversial. In this study, we utilized the monocytic cell line THP-1 as an in vitro latency model for human cytomegalovirus infection (HCMV). Characterization of THP-1 cells by immunofluorescence andWestern blot analysis confirmed the expression of all major ND10 components. THP-1 cells with a stable, individual knockdown of PML, hDaxx or Sp100 were generated. Importantly, depletion of the major ND10 proteins did not prevent the terminal cellular differentiation of THP-1 monocytes. After construction of a recombinant, endotheliotropic human cytomegalovirus expressing IE2-EYFP, we investigated whether the depletion of ND10 proteins affects the onset of viral IE gene expression. While after infection of differentiated, THP-1-derived macrophages as well as during differentiation-induced reactivation from latency an increase in the number of IE-expressing cells was readily detectable in the absence of the major ND10 proteins, no effect was observed in non-differentiated monocytes. We conclude that PML, hDaxx and Sp100 primarily act as cellular restriction factors during lytic HCMV replication and during the dynamic process of reactivation but do not serve as key determinants for the establishment of HCMV latency.
authors.affiliation: Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Schlossgarten 4, 91054 Erlangen, Germany. nadine.wagenknecht@viro.med.uni-erlangen.de.
authors.forename: Nadine
authors.initials: N
authors.lastname: Wagenknecht
authors.affiliation: Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Schlossgarten 4, 91054 Erlangen, Germany. nina.reuter@viro.med.uni-erlangen.de.
authors.forename: Nina
authors.initials: N
authors.lastname: Reuter
authors.affiliation: Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Schlossgarten 4, 91054 Erlangen, Germany. myriam.scherer@viro.med.uni-erlangen.de.
authors.forename: Myriam
authors.initials: M
authors.lastname: Scherer
authors.affiliation: Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Schlossgarten 4, 91054 Erlangen, Germany. aareiche@viro.med.uni-erlangen.de.
authors.forename: Anna
authors.initials: A
authors.lastname: Reichel
authors.affiliation: Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Schlossgarten 4, 91054 Erlangen, Germany. regina.mueller@viro.med.uni-erlangen.de.
authors.forename: Regina
authors.initials: R
authors.lastname: Müller
authors.affiliation: Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Schlossgarten 4, 91054 Erlangen, Germany. thomas.stamminger@viro.med.uni-erlangen.de.
authors.forename: Thomas
authors.initials: T
authors.lastname: Stamminger
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D034961:Antigens, Nuclear / Q000378:metabolism*; D019677:Artificial Gene Fusion; D001324:Autoantigens / Q000378:metabolism*; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D015153:Blotting, Western; D002460:Cell Line; D056970:Co-Repressor Proteins; D003587:Cytomegalovirus / Q000276:immunology / Q000502:physiology*; D020869:Gene Expression Profiling; D055785:Gene Knockdown Techniques; D017930:Genes, Reporter; D054884:Host-Pathogen Interactions; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D008856:Microscopy, Fluorescence; D018832:Molecular Chaperones; D009000:Monocytes / Q000276:immunology / Q000821:virology*; D009687:Nuclear Proteins / Q000378:metabolism*; D000071156:Promyelocytic Leukemia Protein; D013194:Staining and Labeling; D014157:Transcription Factors / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000378:metabolism*; D017735:Virus Latency; D014779:Virus Replication
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D034961:Antigens, Nuclear; D001324:Autoantigens; D001426:Bacterial Proteins; D056970:Co-Repressor Proteins; C106985:DAXX protein, human; D008164:Luminescent Proteins; D018832:Molecular Chaperones; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C054562:yellow fluorescent protein, Bacteria; C066642:SP100 protein, human; C070576:PML protein, human
keywords: ND10; PML; cytomegalovirus; intrinsic immunity; latency; restriction factor