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Human cytomegalovirus long noncoding RNA4.9 regulates viral DNA replication
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Human cytomegalovirus long noncoding RNA4.9 regulates viral DNA replication
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Journal Article
Human cytomegalovirus long noncoding RNA4.9 regulates viral DNA replication
2020
Overview
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.
Publisher
Public Library of Science,Public Library of Science (PLoS)
Subject
/ CRISPR
/ Cytomegalovirus Infections - genetics
/ Cytomegalovirus Infections - microbiology
/ Cytomegalovirus Infections - pathology
/ DNA
/ Gene Expression Regulation, Viral
/ Genes
/ Genomes
/ Genomics
/ Humans
/ Immediate-Early Proteins - genetics
/ Immediate-Early Proteins - metabolism
/ Medicine and Health Sciences
/ Mice
/ Proteins
/ Research and analysis methods
/ RNA
/ RNA, Long Noncoding - genetics
/ Single-stranded DNA-binding protein
/ Viruses
