title: Cellular transcription factor Oct-1 interacts with the Epstein-Barr virus BRLF1 protein to promote disruption of viral latency.
abstract: The Epstein-Barr virus (EBV) latent-to-lytic switch is an essential part of the viral life cycle, but the cellular factors that promote viral reactivation are not well defined. In this report, we demonstrate that the cellular transcription factor Oct-1 cooperates with the EBV immediate-early protein BRLF1 (R, Rta) to induce lytic viral reactivation. We show that cotransfected Oct-1 enhances the ability of BRLF1 to activate lytic gene expression in 293 cells stably infected with a BRLF1-defective EBV mutant (BRLF1-stop) and that Oct-1 increases BRLF1-mediated activation of lytic EBV promoters in reporter gene assays. We find that Oct-1 interacts directly with BRLF1 in vitro and that a mutant BRLF1 protein (the M140A mutant) attenuated for the ability to interact with Oct-1 in vitro is also resistant to Oct-1-mediated transcriptional enhancement in 293 BRLF1-stop cells. Furthermore, we show that cotransfected Oct-1 augments BRLF1 binding to a variety of lytic EBV promoters in chromatin immunoprecipitation (ChIP) assays (including the BZLF1, BMRF1, and SM promoters) and that BRLF1 tethers Oct-1 to lytic EBV promoters. In addition, we demonstrate that an Oct-1 mutant defective in DNA binding (the S335D mutant) still retains the ability to enhance BRLF1 transcriptional effects. Finally, we show that knockdown of endogenous Oct-1 expression reduces the level of constitutive lytic EBV gene expression in both EBV-positive B-cell and EBV-positive epithelial cell lines. These results suggest that Oct-1 acts as a positive regulator of EBV lytic gene expression and that this effect is at least partially mediated through its interaction with the viral protein BRLF1.
authors.affiliation: Department of Oncology (McArdle Laboratory for Cancer Research), University of Wisconsin School of Medicine and Public Health, Madison, WI 53706, USA.
authors.forename: Amanda R
authors.initials: AR
authors.lastname: Robinson
authors.forename: Swee Sen
authors.initials: SS
authors.lastname: Kwek
authors.forename: Stacy R
authors.initials: SR
authors.lastname: Hagemeier
authors.forename: Coral K
authors.initials: CK
authors.lastname: Wille
authors.forename: Shannon C
authors.initials: SC
authors.lastname: Kenney
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D002460:Cell Line; D047369:Chromatin Immunoprecipitation; D004279:DNA, Viral / Q000378:metabolism; D004854:Herpesvirus 4, Human / Q000472:pathogenicity*; D054884:Host-Pathogen Interactions; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism*; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D020125:Mutation, Missense; D050811:Octamer Transcription Factor-1 / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D025941:Protein Interaction Mapping; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014775:Virus Activation; D017735:Virus Latency
chemical_list: C068382:BRLF1 protein, Human herpesvirus 4; D004279:DNA, Viral; D017874:Immediate-Early Proteins; D050505:Mutant Proteins; D050811:Octamer Transcription Factor-1; D015534:Trans-Activators