ctcf prevents the epigenetic drift of ebv latency promoter qpctcf防止ebv延迟启动子qp的表观遗传漂移.pdfVIP

CTCF Prevents the Epigenetic Drift of EBV Latency Promoter Qp 1,2 1 1 1 Italo Tempera , Andreas Wiedmer , Jayaraju Dheekollu , Paul M. Lieberman * 1The Wistar Institute, Philadelphia, Pennsylvania, United States of America, 2 Istituto Pasteur – Fondazione Cenci Bolognetti, Rome, Italy 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 ,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