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PMID: 1326651 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Transient replication of human papillomavirus DNAs.

Journal of virology ·Vol. 66 ·No. 10 ·1992-10-00 ·Pages 5949-58

Del Vecchio AM, Romanczuk H, Howley PM, Baker CC

Abstract

Information on papillomavirus DNA replication has primarily derived from studies with bovine papillomavirus type 1 (BPV-1). Our knowledge of DNA replication of the human papillomaviruses (HPVs) is quite limited, in part because of the lack of a cell culture system capable of supporting the stable replication of HPV DNA. This study demonstrates that the full-length genomic DNAs of HPV types 11 and 18 (HPV-11 and HPV-18), but not HPV-16, are able to replicate transiently after transfection into several different human squamous cell carcinoma cell lines. This system was used to identify the viral cis and trans elements required for DNA replication. The viral origins of replication were localized to a region of the viral long control region. Like BPV-1, E1 and E2 were the only viral factors required in trans for the replication of plasmids containing the origin. Cotransfection of a plasmid expressing the E1 open reading frame (ORF) from HPV-11 with a plasmid that expresses the E2 ORF from HPV-6, HPV-11, HPV-16, or HPV-18 supported the replication of plasmid DNAs containing the origin regions of HPV-11, HPV-16, or HPV-18, indicating that there are functions shared among the corresponding E1 and E2 proteins and origins of these viruses. Although HPV-16 genomic DNA did not replicate by itself under experimental conditions that supported the replication of HPV-11 and HPV-18 genomic DNAs, expression of the HPV-16 early region functions from a strong heterologous promoter supported the replication of a cotransfected plasmid containing the HPV-16 origin of replication. This finding suggests that the inability of the HPV-16 genomic DNA to replicate transiently in the cell lines tested was most likely due to insufficient expression of the viral E1 and/or E2 genes required for DNA replication.

MeSH Terms
Base Sequence Blotting, Southern DNA Replication DNA, Viral/biosynthesis Humans Molecular Sequence Data Open Reading Frames Papillomaviridae/genetics,metabolism Plasmids Promoter Regions, Genetic Transfection Tumor Cells, Cultured
Chemicals
DNA, Viral
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Del Vecchio A M
Laboratory of Tumor Virus Biology, National Cancer Institute, Bethesda, Maryland 20892.
Romanczuk H
Howley P M
Baker C C
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.66.10.5949-5958.1992
Published
1992-10-00
Pages
5949-58
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC241472
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.66.10.5949-5958.1992 PMC Full Text View journal details
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