Human cytomegalovirus long noncoding RNA4.9 regulates viral DNA replication

Tai-Schmiedel, J. et al. (2020) Human cytomegalovirus long noncoding RNA4.9 regulates viral DNA replication. PLoS Pathogens, 16(4), e1008390. (doi: 10.1371/journal.ppat.1008390) (PMID:32294138) (PMCID:PMC7185721)

Abstract

Viruses are known for their extremely compact genomes composed almost entirely of protein-coding genes. Nonetheless, four long noncoding RNAs (lncRNAs) are encoded by human cytomegalovirus (HCMV). Although these RNAs accumulate to high levels during lytic infection, their functions remain largely unknown. Here, we show that HCMV-encoded lncRNA4.9 localizes to the viral nuclear replication compartment, and that its depletion restricts viral DNA replication and viral growth. RNA4.9 is transcribed from the HCMV origin of replication (oriLyt) and forms an RNA-DNA hybrid (R-loop) through its G+C-rich 5’ end, which may be important for the initiation of viral DNA replication. Furthermore, targeting the RNA4.9 promoter with CRISPR-Cas9 or genetic relocalization of oriLyt leads to reduced levels of the viral single-stranded DNA-binding protein (ssDBP), suggesting that the levels of ssDBP are coupled to the oriLyt activity. We further identified a similar, oriLyt-embedded, G+C-rich lncRNA in murine cytomegalovirus (MCMV). These results indicate that HCMV RNA4.9 plays an important role in regulating viral DNA replication, that the levels of ssDBP are coupled to the oriLyt activity, and that these regulatory features may be conserved among betaherpesviruses.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Suarez, Dr Nicolas and Lau, Dr Betty and Kerr, Ms Karen and Davison, Professor Andrew

Creator Roles

Lau, B.: Methodology, Resources
Kerr, K.: Methodology, Resources
Suárez, N.: Data curation, Methodology, Resources
Davison, A. J.: Resources, Writing – original draft, Writing – review and editing

• Authors: Tai-Schmiedel, J., Karniely, S., Lau, B., Ezra, A., Eliyahu, E., Nachshon, A., Kerr, K., Suárez, N., Schwartz, M., Davison, A. J., and Stern-Ginossar, N.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity; College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research
• Journal Name: PLoS Pathogens
• Publisher: Public Library of Science
• ISSN: 1553-7366
• ISSN (Online): 1553-7374
• Published Online: 15 April 2020
• Copyright Holders: Copyright © 2020 Tai-Schmiedel et al.
• First Published: First published in PLoS Pathogens 16(4): e1008390
• Publisher Policy: Reproduced under a Creative Commons License

Project Code	Award No	Project Name	Principal Investigator	Funder's Name	Funder Ref	Lead Dept
65632	1	Genomics of human cytomegalovirus	Andrew Davison	Medical Research Council (MRC)	MC_UU_12014/3	MVLS III - CENTRE FOR VIRUS RESEARCH