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PLoS One. 2014 Dec 30;9(12):e116151. doi: 10.1371/journal.pone.0116151. eCollection 2014.

The transcription map of human papillomavirus type 18 during genome replication in U2OS cells.

Author information

1
Institute of Technology, University of Tartu, Tartu, Tartumaa, Estonia.
2
Icosagen Cell Factory OÜ, Ülenurme vald, Tartumaa, Estonia.
3
Institute of Technology, University of Tartu, Tartu, Tartumaa, Estonia; Icosagen Cell Factory OÜ, Ülenurme vald, Tartumaa, Estonia.
4
Estonian Biocentre, Tartu, Tartumaa, Estonia.
5
Institute of Technology, University of Tartu, Tartu, Tartumaa, Estonia; Icosagen Cell Factory OÜ, Ülenurme vald, Tartumaa, Estonia; Estonian Academy of Sciences, Tallinn, Harjumaa, Estonia.

Abstract

The human osteosarcoma cell line U2OS is useful for studying genome replication of human papillomavirus (HPVs) subtypes that belong to different phylogenetic genera. In this study, we defined the HPV18 transcription map in U2OS cells during transient replication, stable maintenance and vegetative amplification by identifying viral promoter regions, transcription polyadenylation and splicing sites during HPV18 genome replication. Mapping of the HPV18 transcription start sites in U2OS cells revealed five distinct promoter regions (P102, P520, P811, P1193 and P3000). With the exception of P3000, all of these regions have been previously identified during productive HPV18 infection. Collectively, the data suggest that U2OS cells are suitable for studying the replication and transcription properties of HPVs and to serve as a platform for conducting high-throughput drug screens to identify HPV replication inhibitors. In addition, we have identified mRNA species that are initiated from the promoter region P3000, which can encode two E2C regulator proteins that contain only the C-terminal hinge and DNA-binding and dimerization domains of E2. We show that these proteins regulate the initial amplification of HPV18 by modulating viral transcription. Moreover, we show that one of these proteins can act as a transcriptional activator of promoter P102.

PMID:
25548925
PMCID:
PMC4280167
DOI:
10.1371/journal.pone.0116151
[Indexed for MEDLINE]
Free PMC Article

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