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Continuous DNA replication is required for late gene transcription and maintenance of replication compartments in gammaherpesviruses
- Source :
- PLoS Pathogens, PLoS Pathogens, Vol 14, Iss 5, p e1007070 (2018)
- Publication Year :
- 2018
- Publisher :
- Public Library of Science, 2018.
-
Abstract
- Late gene transcription in herpesviruses is dependent on viral DNA replication in cis but the mechanistic basis for this linkage remains unknown. DNA replication results in demethylated DNA, topological changes, removal of proteins and recruitment of proteins to promoters. One or more of these effects of DNA replication may facilitate late gene transcription. Using 5-azacytidine to promote demethylation of DNA, we demonstrate that late gene transcription cannot be rescued by DNA demethylation. Late gene transcription precedes significant increases in DNA copy number, indicating that increased template numbers also do not contribute to the linkage between replication and late gene transcription. By using serial, timed blockade of DNA replication and measurement of late gene mRNA accumulation, we demonstrate that late gene transcription requires ongoing DNA replication. Consistent with these findings, blocking DNA replication led to dissolution of DNA replication complexes which also contain RNA polymerase II and BGLF4, an EBV protein required for transcription of several late genes. These data indicate that ongoing DNA replication maintains integrity of a replication-transcription complex which is required for recruitment and retention of factors necessary for late gene transcription.<br />Author summary Herpesviruses exhibit both latent and lytic replication cycles. Gammaherpesviruses such as Kaposi’s sarcoma-associated herpesvirus and Epstein Barr virus undergo lytic replication when they reactivate from latency. During this process, when infectious virions are produced, an orderly cascade of gene expression occurs. Late lytic genes, which primarily encode structural components of the virion, are only transcribed after replication of the DNA genome has occurred. Unlike early lytic genes, late gene transcription is tightly linked to viral DNA replication; if viral DNA replication is blocked, late gene mRNA accumulation is severely inhibited. The mechanism by which late gene transcription is linked to DNA replication has remained elusive. In this paper we show that a process of continuous DNA replication is required. If one blocks DNA replication, further transcription also ceases, indicating that concurrent DNA replication is required to maintain late transcription. We also show that when DNA replication is blocked, the nuclear complexes in which herpesviruses are replicating dissociate. These replication complexes also serve as factories of viral transcription. When the complexes disperse, proteins required for transcription dissociate from the DNA replication machinery. These data indicate that ongoing DNA replication is necessary to maintain the physical and functional integrity of these structures. Our study provides new insight into this linkage that ensures coordination between viral replication and late gene expression.
- Subjects :
- 0301 basic medicine
Phosphonoacetic Acid
Transcription, Genetic
RNA polymerase II
DNA-Directed DNA Polymerase
Pathology and Laboratory Medicine
Virus Replication
Biochemistry
chemistry.chemical_compound
Transcription (biology)
Medicine and Health Sciences
Enzyme Inhibitors
Promoter Regions, Genetic
lcsh:QH301-705.5
DNA methylation
Genomics
Kaposi's Sarcoma-Associated Herpesvirus
Chromatin
Cell biology
Functional Genomics
Nucleic acids
DNA Demethylation
Medical Microbiology
Viral Pathogens
Viruses
Azacitidine
Epigenetics
Pathogens
DNA modification
Chromatin modification
Research Article
Chromosome biology
lcsh:Immunologic diseases. Allergy
DNA Replication
Gene Expression Regulation, Viral
Herpesviruses
Immunology
DNA transcription
Biology
Microbiology
03 medical and health sciences
Gammaherpesvirinae
Virology
Genetics
Molecular Biology
Gene
Microbial Pathogens
Genes, Immediate-Early
Nucleic Acid Synthesis Inhibitors
Biology and life sciences
DNA replication
Organisms
Promoter
DNA
Viral Replication
Kinetics
030104 developmental biology
DNA demethylation
lcsh:Biology (General)
chemistry
biology.protein
Parasitology
Gene expression
lcsh:RC581-607
DNA viruses
Subjects
Details
- Language :
- English
- ISSN :
- 15537374 and 15537366
- Volume :
- 14
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- PLoS Pathogens
- Accession number :
- edsair.doi.dedup.....63f31be770c7f4ddb13af6160dcf749f