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CTCF prevents the epigenetic drift of EBV latency promoter Qp.
PLoS Pathog 2010; 6(8):e1001048PP

Abstract

The establishment and maintenance of Epstein-Barr Virus (EBV) latent infection requires distinct viral gene expression programs. These gene expression programs, termed latency types, are determined largely by promoter selection, and controlled through the interplay between cell-type specific transcription factors, chromatin structure, and epigenetic modifications. We used a genome-wide chromatin-immunoprecipitation (ChIP) assay to identify epigenetic modifications that correlate with different latency types. We found that the chromatin insulator protein CTCF binds at several key regulatory nodes in the EBV genome and may compartmentalize epigenetic modifications across the viral genome. Highly enriched CTCF binding sites were identified at the promoter regions upstream of Cp, Wp, EBERs, and Qp. Since Qp is essential for long-term maintenance of viral genomes in type I latency and epithelial cell infections, we focused on the role of CTCF in regulating Qp. Purified CTCF bound approximately 40 bp upstream of the EBNA1 binding sites located at +10 bp relative to the transcriptional initiation site at Qp. Mutagenesis of the CTCF binding site in EBV bacmids resulted in a decrease in the recovery of stable hygromycin-resistant episomes in 293 cells. EBV lacking the Qp CTCF site showed a decrease in Qp transcription initiation and a corresponding increase in Cp and Fp promoter utilization at 8 weeks post-transfection. However, by 16 weeks post-transfection, bacmids lacking CTCF sites had no detectable Qp transcription and showed high levels of histone H3 K9 methylation and CpG DNA methylation at the Qp initiation site. These findings provide direct genetic evidence that CTCF functions as a chromatin insulator that prevents the promiscuous transcription of surrounding genes and blocks the epigenetic silencing of an essential promoter, Qp, during EBV latent infection.

Authors+Show Affiliations

The Wistar Institute, Philadelphia, Pennsylvania, United States of America.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20730088

Citation

Tempera, Italo, et al. "CTCF Prevents the Epigenetic Drift of EBV Latency Promoter Qp." PLoS Pathogens, vol. 6, no. 8, 2010, pp. e1001048.
Tempera I, Wiedmer A, Dheekollu J, et al. CTCF prevents the epigenetic drift of EBV latency promoter Qp. PLoS Pathog. 2010;6(8):e1001048.
Tempera, I., Wiedmer, A., Dheekollu, J., & Lieberman, P. M. (2010). CTCF prevents the epigenetic drift of EBV latency promoter Qp. PLoS Pathogens, 6(8), pp. e1001048. doi:10.1371/journal.ppat.1001048.
Tempera I, et al. CTCF Prevents the Epigenetic Drift of EBV Latency Promoter Qp. PLoS Pathog. 2010 Aug 12;6(8):e1001048. PubMed PMID: 20730088.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - CTCF prevents the epigenetic drift of EBV latency promoter Qp. AU - Tempera,Italo, AU - Wiedmer,Andreas, AU - Dheekollu,Jayaraju, AU - Lieberman,Paul M, Y1 - 2010/08/12/ PY - 2009/12/07/received PY - 2010/07/15/accepted PY - 2010/8/24/entrez PY - 2010/8/24/pubmed PY - 2010/12/14/medline SP - e1001048 EP - e1001048 JF - PLoS pathogens JO - PLoS Pathog. VL - 6 IS - 8 N2 - The establishment and maintenance of Epstein-Barr Virus (EBV) latent infection requires distinct viral gene expression programs. These gene expression programs, termed latency types, are determined largely by promoter selection, and controlled through the interplay between cell-type specific transcription factors, chromatin structure, and epigenetic modifications. We used a genome-wide chromatin-immunoprecipitation (ChIP) assay to identify epigenetic modifications that correlate with different latency types. We found that the chromatin insulator protein CTCF binds at several key regulatory nodes in the EBV genome and may compartmentalize epigenetic modifications across the viral genome. Highly enriched CTCF binding sites were identified at the promoter regions upstream of Cp, Wp, EBERs, and Qp. Since Qp is essential for long-term maintenance of viral genomes in type I latency and epithelial cell infections, we focused on the role of CTCF in regulating Qp. Purified CTCF bound approximately 40 bp upstream of the EBNA1 binding sites located at +10 bp relative to the transcriptional initiation site at Qp. Mutagenesis of the CTCF binding site in EBV bacmids resulted in a decrease in the recovery of stable hygromycin-resistant episomes in 293 cells. EBV lacking the Qp CTCF site showed a decrease in Qp transcription initiation and a corresponding increase in Cp and Fp promoter utilization at 8 weeks post-transfection. However, by 16 weeks post-transfection, bacmids lacking CTCF sites had no detectable Qp transcription and showed high levels of histone H3 K9 methylation and CpG DNA methylation at the Qp initiation site. These findings provide direct genetic evidence that CTCF functions as a chromatin insulator that prevents the promiscuous transcription of surrounding genes and blocks the epigenetic silencing of an essential promoter, Qp, during EBV latent infection. SN - 1553-7374 UR - https://www.unboundmedicine.com/medline/citation/20730088/CTCF_prevents_the_epigenetic_drift_of_EBV_latency_promoter_Qp_ L2 - http://dx.plos.org/10.1371/journal.ppat.1001048 DB - PRIME DP - Unbound Medicine ER -