The E8∧E2 Gene Product of Human Papillomavirus Type 16 Represses Early Transcription and Replication but Is Dispensable for Viral Plasmid Persistence in Keratinocytes

Michael J Lace; James R Anson; Gregory S Thomas; Lubomir P Turek; Thomas H Haugen

doi:10.1128/JVI.01481-08

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The E8∧E2 Gene Product of Human Papillomavirus Type 16 Represses Early Transcription and Replication but Is Dispensable for Viral Plasmid Persistence in Keratinocytes

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The E8∧E2 Gene Product of Human Papillomavirus Type 16 Represses Early Transcription and Replication but Is Dispensable for Viral Plasmid Persistence in Keratinocytes

Michael J Lace, James R Anson, Gregory S Thomas, Lubomir P Turek and Thomas H Haugen

Journal of virology, Vol.82(21), pp.10841-10853

11/2008

DOI: 10.1128/JVI.01481-08

PMCID: PMC2573160

PMID: 18753207

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Abstract

A conserved E8 ∧ E2 spliced mRNA is detected in keratinocytes transfected with human papillomavirus type 16 (HPV-16) plasmid DNA. Expression of HPV-16 E8 ∧ E2 (16-E8 ∧ E2) is independent of the major early promoter, P97, and is modulated by both specific splicing events and conserved cis elements in the upstream regulatory region in a manner that differs from transcriptional regulation of other early viral genes. Mutations that disrupt the predicted 16-E8 ∧ E2 message also increase initial HPV-16 plasmid amplification 8- to 15-fold and major early gene (P97) transcription 4- to 5-fold over those of the wild type (wt). Expressing the 16-E8 ∧ E2 gene product from the cytomegalovirus (CMV) promoter represses HPV-16 early gene transcription from P97 in a dose-dependent manner, as detected by RNase protection assays. When expressed from the CMV promoter, 16-E8 ∧ E2 also inhibits the amplification of an HPV-16 plasmid and a heterologous simian virus 40 (SV40) ori plasmid that contains E2 binding sites in cis . In contrast, cotransfections with HPV-16 wt genomes that express physiologic levels of 16-E8 ∧ E2 are sufficient to repress HPV-16 plasmid amplification but are limiting and insufficient for the repression of SV40 amplification. 16-E8 ∧ E2-dependent repression of HPV-16 E1 expression is sufficient to account for this observed inhibition of initial HPV-16 plasmid amplification. Unlike with other papillomaviruses, primary human keratinocytes immortalized by the HPV-16 E8 mutant genome contain more than eightfold-higher levels of unintegrated plasmid than the wt, demonstrating that 16-E8 ∧ E2 limits the viral copy number but is not required for plasmid persistence and maintenance.

Genome and Regulation of Viral Gene Expression

Details

Title: Subtitle: The E8∧E2 Gene Product of Human Papillomavirus Type 16 Represses Early Transcription and Replication but Is Dispensable for Viral Plasmid Persistence in Keratinocytes
Creators: Michael J Lace - Department of Pathology, Veterans Affairs Medical Center
James R Anson - Department of Pathology, Veterans Affairs Medical Center
Gregory S Thomas - Department of Pathology, Veterans Affairs Medical Center
Lubomir P Turek - Department of Pathology, Veterans Affairs Medical Center
Thomas H Haugen - Department of Pathology, Veterans Affairs Medical Center
Resource Type: Journal article
Publication Details: Journal of virology, Vol.82(21), pp.10841-10853
DOI: 10.1128/JVI.01481-08
PMID: 18753207
PMCID: PMC2573160
NLM abbreviation: J Virol
ISSN: 0022-538X
eISSN: 1098-5514
Publisher: American Society for Microbiology (ASM)
Language: English
Date published: 11/2008
Academic Unit: Pathology; Medicine Administration
Record Identifier: 9984047880502771

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