Format

Send to

Choose Destination
PLoS One. 2015 Jul 1;10(7):e0131675. doi: 10.1371/journal.pone.0131675. eCollection 2015.

The Cell Cycle Timing of Human Papillomavirus DNA Replication.

Author information

1
University of Tartu, Institute of Technology Department of Biomedical Technology, Nooruse 1, 50411, Tartu, Estonia.
2
University of Tartu, Institute of Technology Department of Biomedical Technology, Nooruse 1, 50411, Tartu, Estonia; Estonian Academy of Sciences, Kohtu 6, 10130, Tallinn, Estonia; Icosagen Cell Factory OÜ, Nooruse 9, 50411, Tartu, Estonia.

Abstract

Viruses manipulate the cell cycle of the host cell to optimize conditions for more efficient viral genome replication. One strategy utilized by DNA viruses is to replicate their genomes non-concurrently with the host genome; in this case, the viral genome is amplified outside S phase. This phenomenon has also been described for human papillomavirus (HPV) vegetative genome replication, which occurs in G2-arrested cells; however, the precise timing of viral DNA replication during initial and stable replication phases has not been studied. We developed a new method to quantitate newly synthesized DNA levels and used this method in combination with cell cycle synchronization to show that viral DNA replication is initiated during S phase and is extended to G2 during initial amplification but follows the replication pattern of cellular DNA during S phase in the stable maintenance phase. E1 and E2 protein overexpression changes the replication time from S only to both the S and G2 phases in cells that stably maintain viral episomes. These data demonstrate that the active synthesis and replication of the HPV genome are extended into the G2 phase to amplify its copy number and the duration of HPV genome replication is controlled by the level of the viral replication proteins E1 and E2. Using the G2 phase for genome amplification may be an important adaptation that allows exploitation of changing cellular conditions during cell cycle progression. We also describe a new method to quantify newly synthesized viral DNA levels and discuss its benefits for HPV research.

PMID:
26132923
PMCID:
PMC4489393
DOI:
10.1371/journal.pone.0131675
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Public Library of Science Icon for PubMed Central
Loading ...
Support Center