Genetic analysis of high-risk E6 in episomal maintenance of human papillomavirus genomes in primary human keratinocytes

Regina B. Park, Elliot Androphy

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

Papillomaviruses possess small DNA genomes that encode five early (E) proteins. Transient DNA replication requires activities of the E1 and E2 proteins and a DNA segment containing their binding sites. The E6 and E7 proteins of cancer-associated human papillomavirus (HPV) transform cells in culture. Recent reports have shown that E6 and E7 are necessary for episomal maintenance of HPV in primary keratinocytes. The functions of E6 necessary for viral replication have not been determined, and to address this question we used a recently developed transfection system based on HPV31. To utilize a series of HPV16 E6 mutations, HPV31 E6 was replaced by its HPV16 counterpart. This chimeric genome was competent for both transient and stable replication in keratinocytes. Four HPV16 E6 mutations that do not stimulate p53 degradation were unable to support stable viral replication, suggesting this activity may be necessary for episomal maintenance. E7 has also been shown to be essential for episomal maintenance of the HPV31 genome. A point mutation in the Rb binding motif of HPV E7 has been reported to render HPV31 unable to stably replicate. Interestingly, HPV31 genomes harboring two of the three p53 degradation-defective E6 mutations combined with this E7 mutation were maintained as replicating episomes. These findings imply that the balance between E6 and E7 functions in infected cells is critical for episomal maintenance of high-risk HPV genomes. This model will be useful to dissect the activities of E6 and E7 necessary for viral DNA replication.

Original languageEnglish (US)
Pages (from-to)11359-11364
Number of pages6
JournalJournal of Virology
Volume76
Issue number22
DOIs
StatePublished - 2002
Externally publishedYes

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Papillomaviridae
keratinocytes
Human Genome
Keratinocytes
genetic techniques and protocols
Genome
Mutation
genome
Maintenance
mutation
DNA Replication
DNA replication
virus replication
Proteins
DNA
Viral DNA
Point Mutation
degradation
proteins
Transfection

ASJC Scopus subject areas

  • Immunology

Cite this

Genetic analysis of high-risk E6 in episomal maintenance of human papillomavirus genomes in primary human keratinocytes. / Park, Regina B.; Androphy, Elliot.

In: Journal of Virology, Vol. 76, No. 22, 2002, p. 11359-11364.

Research output: Contribution to journalArticle

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