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
The human osteosarcoma cell line U2OS is useful for studying genome replicationof human papillomavirus (HPVs) subtypes that belong to different phylogeneticgenera. In this study, we defined the HPV18 transcription map in U2OS cells duringtransient replication, stable maintenance and vegetative amplification by identifyingviral promoter regions, transcription polyadenylation and splicing sites duringHPV18 genome replication. Mapping of the HPV18 transcription start sites in U2OScells revealed five distinct promoter regions (P102, P520, P811, P1193 and P3000). Withthe exception of P3000, all of these regions have been previously identified duringproductive HPV18 infection. Collectively, the data suggest that U2OS cells aresuitable for studying the replication and transcription properties of HPVs and toserve as a platform for conducting high-throughput drug screens to identify HPVreplication inhibitors. In addition, we have identified mRNA species that are initiatedfrom the promoter region P3000, which can encode two E2C regulator proteins thatcontain only the C-terminal hinge and DNA-binding and dimerization domains ofE2. We show that these proteins regulate the initial amplification of HPV18 bymodulating viral transcription. Moreover, we show that one of these proteins can actas a transcriptional activator of promoter P102
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