title: Transient replication of human papillomavirus DNAs.
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.
authors.affiliation: Laboratory of Tumor Virus Biology, National Cancer Institute, Bethesda, Maryland 20892.
authors.forename: A M
authors.initials: AM
authors.lastname: Del Vecchio
authors.forename: H
authors.initials: H
authors.lastname: Romanczuk
authors.forename: P M
authors.initials: PM
authors.lastname: Howley
authors.forename: C C
authors.initials: CC
authors.lastname: Baker
mesh_terms: D001483:Base Sequence; D015139:Blotting, Southern; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis*; D006801:Humans; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D027383:Papillomaviridae / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D004279:DNA, Viral