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PMID: 24068939 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Inactivation of intergenic enhancers by EBNA3A initiates and maintains polycomb signatures across a chromatin domain encoding CXCL10 and CXCL9.

PLoS pathogens ·Vol. 9 ·No. 9 ·2013-09-00 ·Pages e1003638

Harth-Hertle ML, Scholz BA, Erhard F, Glaser LV, Dölken L, Zimmer R, Kempkes B

Abstract

Epstein-Barr virus (EBV) causes a persistent infection in human B cells by establishing specific transcription programs to control B cell activation and differentiation. Transcriptional reprogramming of EBV infected B cells is predominantly driven by the action of EBV nuclear antigens, among them the transcriptional repressor EBNA3A. By comparing gene expression profiles of wt and EBNA3A negative EBV infected B cells, we have previously identified a broad array of cellular genes controlled by EBNA3A. We now find that genes repressed by EBNA3A in these cells are significantly enriched for the repressive histone mark H3K27me3, which is installed by Polycomb group (PcG) proteins. This PcG-controlled subset of genes also carries H3K27me3 marks in a variety of other tissues, suggesting that the commitment to PcG silencing is an intrinsic feature of these gene loci that can be used by EBNA3A. In addition, EBNA3A targets frequently reside in co-regulated gene clusters. To study the mechanism of gene repression by EBNA3A and to evaluate the relative contribution of PcG proteins during this process, we have selected the genomic neighbors CXCL10 and CXCL9 as a model for co-repressed and PcG-controlled genes. We show that EBNA3A binds to CBF1 occupied intergenic enhancers located between CXCL10 and CXCL9 and displaces the transactivator EBNA2. This impairs enhancer activity, resulting in a rapid transcriptional shut-down of both genes in a CBF1-dependent manner and initiation of a delayed gain of H3K27me3 marks covering an extended chromatin domain. H3K27me3 marks increase gradually and are maintained by EBNA3A. Our study provides direct evidence that repression by EBNA3A requires CBF1 and that EBNA3A and EBNA2 compete for access to CBF1 at identical genomic sites. Most importantly, our results demonstrate that transcriptional silencing by EBNA3A precedes the appearance of repressive PcG marks and indicate that both events are triggered by loss of enhancer activity.

MeSH Terms
B-Lymphocytes/immunology,metabolism,virology Cell Line Cellular Reprogramming Chemokine CXCL10/genetics,metabolism Chemokine CXCL9/genetics,metabolism Chromatin Assembly and Disassembly DNA, Intergenic/metabolism Enhancer Elements, Genetic Epstein-Barr Virus Infections/immunology,metabolism,virology Epstein-Barr Virus Nuclear Antigens/genetics,metabolism Herpesvirus 4, Human/immunology,metabolism Host-Pathogen Interactions Humans Immunoglobulin J Recombination Signal Sequence-Binding Protein/genetics,metabolism Models, Biological Mutation Polycomb-Group Proteins/metabolism Recombinant Proteins/metabolism Repressor Proteins/genetics,metabolism Trans-Activators/metabolism Viral Proteins/metabolism
Chemicals
CXCL10 protein, human CXCL9 protein, human Chemokine CXCL10 Chemokine CXCL9 DNA, Intergenic EBNA-2 protein, Human herpesvirus 4 EBNA-3A antigen Epstein-Barr Virus Nuclear Antigens Immunoglobulin J Recombination Signal Sequence-Binding Protein Polycomb-Group Proteins RBPJ protein, human Recombinant Proteins Repressor Proteins Trans-Activators Viral Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Harth-Hertle Marie L
Department of Gene Vectors, Helmholtz Center Munich, German Research Center for Environmental Health, Munich, Germany.
Scholz Barbara A
Erhard Florian
Glaser Laura V
Dölken Lars
Zimmer Ralf
Kempkes Bettina
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Article Info
Journal
PLoS pathogens
Abbr.
PLoS Pathog
ISSN
1553-7374
DOI
10.1371/journal.ppat.1003638
Published
2013-09-00
Epub
2013-00-19
Pages
e1003638
Language
English
Region
United States
NLM ID
101238921
PMCID
PMC3777872
Subset
IM
Grants
Medical Research Council · G1002523 · United Kingdom
External Links
PubMed source DOI: 10.1371/journal.ppat.1003638 PMC Full Text View journal details
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