Single-genome analysis reveals heterogeneous association of the herpes simplex virus genome with H3K27me2 and the reader PHF20L1 following infection of human fibroblasts

Francois, A. K. et al. (2024) Single-genome analysis reveals heterogeneous association of the herpes simplex virus genome with H3K27me2 and the reader PHF20L1 following infection of human fibroblasts. mBio, (doi: 10.1128/mbio.03278-23) (PMID:32753500) (PMCID:PMC7407090) (Early Online Publication)

Abstract

The fate of herpesvirus genomes following entry into different cell types is thought to regulate the outcome of infection. For the Herpes simplex virus 1 (HSV-1), latent infection of neurons is characterized by association with repressive heterochromatin marked with Polycomb silencing-associated lysine 27 methylation on histone H3 (H3K27me). However, whether H3K27 methylation plays a role in repressing lytic gene expression in non-neuronal cells is unclear. To address this gap in knowledge, and with consideration that the fate of the viral genome and outcome of HSV-1 infection could be heterogeneous, we developed an assay to quantify the abundance of histone modifications within single viral genome foci of infected fibroblasts. Using this approach, combined with bulk epigenetic techniques, we were unable to detect any role for H3K27me3 during HSV-1 lytic infection of fibroblasts. By contrast, we could detect the lesser studied H3K27me2 on a subpopulation of viral genomes, which was consistent with a role for H3K27 demethylases in promoting lytic gene expression. In addition, viral genomes co-localized with the H3K27me2 reader protein PHF20L1, and this association was enhanced by inhibition of the H3K27 demethylases UTX and JMJD3. Notably, targeting of H3K27me2 to viral genomes was enhanced following infection with a transcriptionally defective virus in the absence of Promyelocytic leukemia nuclear bodies. Collectively, these studies implicate a role for H3K27me2 in fibroblast-associated HSV genome silencing in a manner dependent on genome sub-nuclear localization and transcriptional activity.

• Item Type: Articles
• Additional Information: This work was supported by The Owens Family Foundation (ARC), National Institutes of Health grants T32GM008136 (SAD and AKF), T32AI007046 (ALW), F32CA260116 (JH) and U54CA274499(DK), MRC awards MC_UU_12014/5 and MC_UU_00034/2 (CB), American Cancer Society award RSG-21–031-01-DMC (SBR).
• Status: Early Online Publication
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Boutell, Dr Chris and McFarlane, Mr Steven

Creator Roles

McFarlane, S.: Investigation, Resources
Boutell, C.: Funding acquisition, Resources, Supervision

• Authors: Francois, A. K., Rohani, A., Loftus, M., Dochnal, S., Hrit, J., McFarlane, S., Whitford, A., Lewis, A., Krakowiak, P., Boutell, C., Rothbart, S. B., Kashatus, D., and Cliffe, A. R.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: mBio
• Publisher: American Society for Microbiology
• ISSN: 2161-2129
• ISSN (Online): 2150-7511
• Published Online: 27 February 2024
• Copyright Holders: Copyright © 2024 Francois et al.
• First Published: First published in mBio 2024
• Publisher Policy: Reproduced under a Creative Commons licence

Project Code	Award No	Project Name	Principal Investigator	Funder's Name	Funder Ref	Lead Dept
172630009		Analysing the Role of Ubiquitin and Ubiquitin-like Modification During Herpesvirus Infection and Host Immunity (Programme 6)	Chris Boutell	Medical Research Council (MRC)	MC_UU_12014/5	III - Centre for Virus Research