title: Mapping of betapapillomavirus human papillomavirus 5 transcription and characterization of viral-genome replication function.
abstract: Betapapillomavirus replication and transcription have not been studied in detail because of a lack of suitable cellular systems supporting human papillomavirus (HPV) genome replication. We have recently shown that the human osteosarcoma cell line U2OS provides a useful environment for the genome replication of many different HPVs, including the betapapillomaviruses HPV5 and HPV8. Using mutational analysis and complementation assay, we demonstrated herein that the viral early proteins E1 and E2 are viral transfactors that are necessary and sufficient for HPV5 genome replication. We also identified four HPV5 early promoter regions with transcription start sites (TSSs) at nucleotides (nt) 184/191, 460, 840, and 1254, respectively, and the HPV late promoter with a TSS at nt 7640. In addition, we mapped the HPV5 early polyadenylation cleavage sites via 3' rapid amplification of cDNA ends (3'RACE) to nt 4457 and 4475. In total, 14 different viral mRNA species, originating from the HPV5 genome, were mapped in U2OS cells during transient and stable replication. The main splicing donor and acceptor sites identified herein are consistent with the data previously obtained in HPV5-positive skin lesions. In addition, we identified novel E8 open reading frame (ORF)-containing transcripts (E8^E1C and E8^E2C) expressed from the HPV5 genome. Similar to several other papillomaviruses, the product of the E8^E2C mRNA acts as a repressor of viral genome replication.
authors.affiliation: Estonian Biocenter, Tartu, Estonia.
authors.forename: Eve
authors.initials: E
authors.lastname: Sankovski
authors.forename: Andres
authors.initials: A
authors.lastname: Männik
authors.forename: Jelizaveta
authors.initials: J
authors.lastname: Geimanen
authors.forename: Ene
authors.initials: E
authors.lastname: Ustav
authors.forename: Mart
authors.initials: M
authors.lastname: Ustav
mesh_terms: D052686:Betapapillomavirus / Q000235:genetics* / Q000502:physiology; D045744:Cell Line, Tumor; D002874:Chromosome Mapping; D015967:Gene Expression Regulation, Viral; D016679:Genome, Viral; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism; D030361:Papillomavirus Infections / Q000821:virology; D011401:Promoter Regions, Genetic; D024363:Transcription Initiation Site; D014158:Transcription, Genetic; D014779:Virus Replication
chemical_list: D009856:Oncogene Proteins, Viral