PMID- 30996085 OWN - NLM STAT- MEDLINE DCOM- 20200528 LR - 20200528 IS - 1098-5514 (Electronic) IS - 0022-538X (Print) IS - 0022-538X (Linking) VI - 93 IP - 13 DP - 2019 Jul 1 TI - The CCCTC Binding Factor, CTRL2, Modulates Heterochromatin Deposition and the Establishment of Herpes Simplex Virus 1 Latency In Vivo. LID - 10.1128/JVI.00415-19 [doi] LID - e00415-19 AB - The cellular insulator protein CTCF plays a role in herpes simplex virus 1 (HSV-1) latency through the establishment and regulation of chromatin boundaries. We previously found that the CTRL2 regulatory element downstream from the latency-associated transcript (LAT) enhancer was bound by CTCF during latency and underwent CTCF eviction at early times postreactivation in mice latently infected with 17syn+ virus. We also showed that CTRL2 was a functional enhancer-blocking insulator in both epithelial and neuronal cell lines. We hypothesized that CTRL2 played a direct role in silencing lytic gene expression during the establishment of HSV-1 latency. To test this hypothesis, we used a recombinant virus with a 135-bp deletion spanning only the core CTRL2 insulator domain (DeltaCTRL2) in the 17syn+ background. Deletion of CTRL2 resulted in restricted viral replication in epithelial cells but not neuronal cells. Following ocular infection, mouse survival decreased in the DeltaCTRL2-infected cohort, and we found a significant decrease in the number of viral genomes in mouse trigeminal ganglia (TG) infected with DeltaCTRL2, indicating that the CTRL2 insulator was required for the efficient establishment of latency. Immediate early (IE) gene expression significantly increased in the number of ganglia infected with DeltaCTRL2 by 31 days postinfection relative to the level with 17syn+ infection, indicating that deletion of the CTRL2 insulator disrupted the organization of chromatin domains during HSV-1 latency. Finally, chromatin immunoprecipitation with high-throughput sequencing (ChIP-seq) analyses of TG from DeltaCTRL2-infected mice confirmed that the distribution of the repressive H3K27me3 (histone H3 trimethylated at K27) mark on the DeltaCTRL2 recombinant genomes was altered compared to that of the wild type, indicating that the CTRL2 site modulates the repression of IE genes during latency.IMPORTANCE It is becoming increasingly clear that chromatin insulators play a key role in the transcriptional control of DNA viruses. The gammaherpesviruses Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV) utilize chromatin insulators to order protein recruitment and dictate the formation of three-dimensional DNA loops that spatially control transcription and latency. The contribution of chromatin insulators in alphaherpesvirus transcriptional control is less well understood. The work presented here begins to bridge that gap in knowledge by showing how one insulator site in HSV-1 modulates lytic gene transcription and heterochromatin deposition as the HSV-1 genome establishes latency. CI - Copyright (c) 2019 American Society for Microbiology. FAU - Washington, Shannan D AU - Washington SD AD - Department of Pharmacology and Experimental Therapeutics, Louisiana State University Health Sciences Center, New Orleans, Louisiana, USA. FAU - Singh, Pankaj AU - Singh P AD - Department of Pharmacology and Experimental Therapeutics, Louisiana State University Health Sciences Center, New Orleans, Louisiana, USA. AD - Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin, USA. FAU - Johns, Richard N AU - Johns RN AD - Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida, USA. FAU - Edwards, Terri G AU - Edwards TG AD - Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida, USA. FAU - Mariani, Michael AU - Mariani M AD - Department of Biomedical and Health Sciences, University of Vermont, Burlington, Vermont, USA. FAU - Frietze, Seth AU - Frietze S AUID- ORCID: 0000-0003-4058-3661 AD - Department of Biomedical and Health Sciences, University of Vermont, Burlington, Vermont, USA. AD - University of Vermont Cancer Center, Burlington, Vermont, USA. FAU - Bloom, David C AU - Bloom DC AD - Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida, USA. FAU - Neumann, Donna M AU - Neumann DM AD - Department of Pharmacology and Experimental Therapeutics, Louisiana State University Health Sciences Center, New Orleans, Louisiana, USA dneumann3@wisc.edu. AD - Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin, USA. LA - eng GR - R01 AI048633/AI/NIAID NIH HHS/United States GR - R01 AI134807/AI/NIAID NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20190614 PL - United States TA - J Virol JT - Journal of virology JID - 0113724 RN - 0 (CCCTC-Binding Factor) RN - 0 (Chromatin) RN - 0 (Ctcf protein, mouse) RN - 0 (Heterochromatin) SB - IM MH - Animals MH - CCCTC-Binding Factor/genetics/*metabolism MH - Chromatin/metabolism MH - Chromatin Assembly and Disassembly MH - Chromatin Immunoprecipitation MH - Disease Models, Animal MH - Epigenomics MH - Eye Infections/virology MH - Ganglia/virology MH - Gene Expression Regulation, Viral MH - Gene Silencing MH - Genome, Viral MH - Herpes Simplex/virology MH - Herpesvirus 1, Human/genetics/*metabolism MH - Herpesvirus 4, Human/physiology MH - Herpesvirus 8, Human/physiology MH - Heterochromatin/*metabolism MH - Mice MH - Mice, Inbred BALB C MH - Mice, Knockout MH - Virus Activation MH - Virus Latency/*physiology MH - Virus Replication PMC - PMC6580944 OTO - NOTNLM OT - CTCF OT - CTRL2 OT - HSV-1 OT - chromatin remodeling OT - epigenetics OT - insulator OT - latency OT - mouse ocular model EDAT- 2019/04/19 06:00 MHDA- 2020/05/29 06:00 PMCR- 2019/12/14 CRDT- 2019/04/19 06:00 PHST- 2019/03/08 00:00 [received] PHST- 2019/04/04 00:00 [accepted] PHST- 2019/04/19 06:00 [pubmed] PHST- 2020/05/29 06:00 [medline] PHST- 2019/04/19 06:00 [entrez] PHST- 2019/12/14 00:00 [pmc-release] AID - JVI.00415-19 [pii] AID - 00415-19 [pii] AID - 10.1128/JVI.00415-19 [doi] PST - epublish SO - J Virol. 2019 Jun 14;93(13):e00415-19. doi: 10.1128/JVI.00415-19. Print 2019 Jul 1.