EBNA2 Drives Formation of New Chromosome Binding Sites and Target Genes for B-Cell Master Regulatory Transcription Factors RBP-jκ and EBF1

TY - JOUR

T1 - EBNA2 Drives Formation of New Chromosome Binding Sites and Target Genes for B-Cell Master Regulatory Transcription Factors RBP-jκ and EBF1

AU - Lu, Fang

AU - Chen, Horng-Shen

AU - Kossenkov, Andrew V

AU - DeWispeleare, Karen

AU - Won, Kyoung-Jae

AU - Lieberman, Paul M

PY - 2016/1

Y1 - 2016/1

N2 - 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.

AB - 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.

KW - B-Lymphocytes/virology

KW - Blotting, Western

KW - Cell Transformation, Viral/genetics

KW - Chromatin Immunoprecipitation

KW - Epstein-Barr Virus Nuclear Antigens/genetics

KW - Gene Expression Regulation, Viral/genetics

KW - High-Throughput Nucleotide Sequencing

KW - Humans

KW - Immunoglobulin J Recombination Signal Sequence-Binding Protein/genetics

KW - Oligonucleotide Array Sequence Analysis

KW - Polymerase Chain Reaction

KW - Trans-Activators/genetics

KW - Viral Proteins/genetics

U2 - 10.1371/journal.ppat.1005339

DO - 10.1371/journal.ppat.1005339

M3 - Journal article

C2 - 26752713

SN - 1553-7366

VL - 12

SP - e1005339

JO - P L o S Pathogens

JF - P L o S Pathogens

IS - 1

ER -