The fidelity of HPV16 E1/E2-mediated DNA replication.
Human papillomaviruses (HPV) are causative agents in a variety of human diseases; for example over 99% of cervical carcinomas contain HPV DNA sequences. Often in cervical carcinoma the HPV genome is integrated into the host genome resulting in unregulated expression of the viral transforming proteins E6 and E7. Therefore viral integration is a step toward HPV-induced carcinogenesis. Integration of the HPV genome could occur following double-strand DNA breaks that could arise during viral DNA replication. We investigated the fidelity of HPV 16 E1- and E2-mediated DNA replication of non-damaged and UVC-damaged templates in a variety of cell lines with different genetic backgrounds; C33a (derived from an HPV-negative cervical carcinoma), XP30RO (deficient in the by-pass polymerase eta (poleta)), XP30eta (expressing a restored wild-type poleta), XP12RO (nucleotide excision repair defective), and MRC5 (derived from a 14-week-old human fetus). The results demonstrate that the fidelity of E1- and E2-mediated DNA replication is reflective of the genetic background in which the assays are carried out. For example, restoring poleta to the XP30 cell line results in a 3-fold drop in the number of mutants obtained following replication of a UVC-damaged template. A relatively high percentage of the mutant-replicated molecules arise as a result of genetic rearrangement. This is the first time such studies have been carried out with an HPV replication system, and the results are discussed in the context of the HPV life cycle and what is known about HPV genomes in human cancers.
Taylor
Ewan R
ER
Institute of Comparative Medicine, Department of Veterinary Pathology, University of Glasgow, Garscube Estate, Bearsden Road, Glasgow G61 1QH, Scotland.
Dornan
Edward S
ES
Boner
Winifred
W
Connolly
Julie A
JA
McNair
Shona
S
Kannouche
Patricia
P
Lehmann
A R
AR
Morgan
Iain M
IM
D015139:Blotting, Southern; D002460:Cell Line; D045744:Cell Line, Tumor; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D004261:DNA Replication; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000378:metabolism; D016679:Genome, Viral; D006801:Humans; D016296:Mutagenesis; D009154:Mutation; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D010957:Plasmids / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014162:Transfection; D014466:Ultraviolet Rays
D004268:DNA-Binding Proteins; C093478:E1 protein, Human papillomavirus 16; C054855:E2 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D004247:DNA