title: EBNA2 Drives Formation of New Chromosome Binding Sites and Target Genes for B-Cell Master Regulatory Transcription Factors RBP-jκ and EBF1.
abstract: Epstein-Barr Virus (EBV) transforms resting B-lymphocytes into proliferating lymphoblasts to establish latent infections that can give rise to malignancies. We show here that EBV-encoded transcriptional regulator EBNA2 drives the cooperative and combinatorial genome-wide binding of two master regulators of B-cell fate, namely EBF1 and RBP-jκ. Previous studies suggest that these B-cell factors are statically bound to target gene promoters. In contrast, we found that EBNA2 induces the formation of new binding for both RBP-jκ and EBF1, many of which are in close physical proximity in the cellular and viral genome. These newly induced binding sites co-occupied by EBNA2-EBF1-RBP-jκ correlate strongly with transcriptional activation of linked genes that are important for B-lymphoblast function. Conditional expression or repression of EBNA2 leads to a rapid alteration in RBP-jκ and EBF1 binding. Biochemical and shRNA depletion studies provide evidence for cooperative assembly at co-occupied sites. These findings reveal that EBNA2 facilitate combinatorial interactions to induce new patterns of transcription factor occupancy and gene programming necessary to drive B-lymphoblast growth and survival.
authors.affiliation: The Wistar Institute, Philadelphia, Pennsylvania, United States of America.
authors.forename: Fang
authors.initials: F
authors.lastname: Lu
authors.affiliation: The Wistar Institute, Philadelphia, Pennsylvania, United States of America.
authors.forename: Horng-Shen
authors.initials: HS
authors.lastname: Chen
authors.affiliation: The Wistar Institute, Philadelphia, Pennsylvania, United States of America.
authors.forename: Andrew V
authors.initials: AV
authors.lastname: Kossenkov
authors.affiliation: The Wistar Institute, Philadelphia, Pennsylvania, United States of America.
authors.forename: Karen
authors.initials: K
authors.lastname: DeWispeleare
authors.affiliation: The Institute for Diabetes Obesity and Metabolism, Department of Genetics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.
authors.forename: Kyoung-Jae
authors.initials: KJ
authors.lastname: Won
authors.affiliation: The Wistar Institute, Philadelphia, Pennsylvania, United States of America.
authors.forename: Paul M
authors.initials: PM
authors.lastname: Lieberman
mesh_terms: D001402:B-Lymphocytes / Q000821:virology*; D015153:Blotting, Western; D002472:Cell Transformation, Viral / Q000235:genetics*; D047369:Chromatin Immunoprecipitation; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics*; D015967:Gene Expression Regulation, Viral / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D051819:Immunoglobulin J Recombination Signal Sequence-Binding Protein / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D015534:Trans-Activators / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics*
chemical_list: C081976:EBF1 protein, human; C402641:EBNA-2 protein, Human herpesvirus 4; D019309:Epstein-Barr Virus Nuclear Antigens; D051819:Immunoglobulin J Recombination Signal Sequence-Binding Protein; C497570:RBPJ protein, human; D015534:Trans-Activators; D014764:Viral Proteins