Department of Microbiology, University of Colorado School of Medicine, Aurora, Colorado, United States of America.
PLoS One. 2012;7(1):e29410. doi: 10.1371/journal.pone.0029410. Epub 2012 Jan 18.
Regulation of viral transcription by chromatin structure has emerged as a fundamental determinant in the establishment of lytic and latent herpesvirus infections. The Polycomb group (PcG) of epigenetic repressors promotes heterochromatin formation by trimethylating histone H3 on lysine-27 (H3K27me3) and regulates development, stem cell renewal and differentiation and the cell cycle. These cellular processes are tightly coupled to the molecular switch between lytic and latent herpesvirus infections. Using chromatin immunoprecipitation analysis, we observed enrichment of H3K27me3 at the major immediate-early (MIE) locus of murine cytomegalovirus (MCMV) very early following infection of permissive fibroblasts. As lytic replication progressed, we observed a loss of H3K27me3 enrichment concomitant with the appearance of H3K4me3. However, late during infection, as viral replication centers are established, we observed a significant increase in PcG protein association with chromatin. Additionally, in co-immunofluorescence assays using confocal microscopy, we detected strong enrichments for PcG protein within the viral replication compartment, suggesting an association between viral DNA synthesis machinery and PcG proteins. Together, our results suggest a novel, dynamic interaction between PcG epigenetic repressors and MCMV genomes.
染色质结构对病毒转录的调控已成为决定疱疹病毒裂解和潜伏感染的基本因素。表观遗传抑制剂 Polycomb 组蛋白通过将组蛋白 H3 赖氨酸 27 三甲基化(H3K27me3)促进异染色质形成,并调节发育、干细胞更新和分化以及细胞周期。这些细胞过程与疱疹病毒裂解和潜伏感染之间的分子开关紧密耦合。通过染色质免疫沉淀分析,我们观察到在允许的成纤维细胞感染后,鼠巨细胞病毒(MCMV)的主要早期(MIE)基因座上 H3K27me3 的富集非常早。随着裂解复制的进展,我们观察到 H3K27me3 富集的丧失伴随着 H3K4me3 的出现。然而,在感染后期,当建立病毒复制中心时,我们观察到 PcG 蛋白与染色质的结合显著增加。此外,在使用共聚焦显微镜的共免疫荧光分析中,我们在病毒复制区检测到 PcG 蛋白的强烈富集,表明病毒 DNA 合成机制与 PcG 蛋白之间存在关联。总之,我们的结果表明 PcG 表观遗传抑制剂与 MCMV 基因组之间存在一种新的动态相互作用。