Using a focus assay we have shown that the entire human papillomavirus type 16 (HPV-16) genome is capable of cooperating with an activated ras oncogene to transform secondary rat embryo fibroblast (REF) cells as indicated by focus formation of unselected cells. However, this assay failed to detect any similar activity with either the whole HPV-6 genome or with subgenomic fragments. In contrast, transformed colonies appeared when G418 (geneticin)-resistant colonies were selected after cotransfection with activated ras DNA and either the entire HPV-6 genome or subgenomic fragments containing the E6/E7 open reading frames (ORFs) of HPV-6 or HPV-16. The transformation assessment was based on the development of a ras-transformed appearance in G418-resistant colonies. The appearance of this morphology did not imply the ability of transformed cells to produce colonies in semi-solid agarose (anchorage-independent growth), and extended culture for about 10 to 20 population doublings was necessary before transfected cells exhibited anchorage-independent growth. Transformation of REF cells was not observed with the E5 ORF of HPV-16 under the control of an exogenous promoter (the long terminal repeat of Rous sarcoma virus) in cooperation with activated ras DNA. No transformation was observed using an activated myc oncogene with either HPV-6 or HPV-16 DNA.
CampionM. J.,
McCanceD. I.,
CuzickJ.,
SingerA.1983; The progressive potential of mild cervical atypia: a prospective cytological, colposcopic and virological study. Lancet ii:237–240
DürstM.,
GissmannL.,
IkenbergH.,
Zur HausenH.1983; A papillomavirus DNA from a cervical carcinoma and its prevalence in cancer biopsy samples from different geographic regions. Proceedings of the National Academy of SciencesU.S.A 80:3812–3815
HaugenT. H.,
CripeT. P.,
GinderG. D.,
KarinM.,
TurekL. P.1987; Transactivation of an upstream early gene promoter of bovine papillomavirus-1 by a product of the viral E2 gene. EMBO Journal 6:145–152
KandaT.,
WatanabeS.,
YoshiikeK.1987; Human papillomavirus type 16 transformation of rat 3Y1 cells. Japanese Journal of Cancer Research (Gann) 78:103–108
KelekarA.,
ColeM. D.1986; Tumorigenicity of fibroblast lines expressing the adenovirus Ela cellular p53, or normal c-myc genes. Molecular and Cellular Biology 6:7–14
MatlashewskiG.,
SchneiderJ.,
BanksL.,
JonesN.,
MurrayA.,
CrawfordL.1987; Human papillomavirus type 16 DNA cooperates with activated ras in transforming primary cells. EMBO Journal 6:1741–1746
MeneguzziG.,
BinétruyM.,
CuzinF.1984; Plasmidial maintenance in rodent fibroblasts of a BPVl-pBR322 shuttle vector without immediately apparent oncogenic transformation of the recipient cells. EMBO Journal 3:365–371
ParadaL. F.,
TabinC. J.,
ShihC.,
WeinbergR. A.1982; Human EJ bladder carcinoma oncogene is a homologue of Harvey sarcoma virus ras gene. Nature, London 297:474–478
PhelpsW. C.,
YeeC. L.,
MungerK.,
HowleyP. M.1988; The human papillomavirus type 16 E7 gene encodes transactivation and transformation functions similar to those of adenovirus E1A. Cell 53:539–547
SouthernP. J.,
BergP.1982; Transformation of mammalian cells to antibiotic resistance with a bacterial gene under the control of the SV40 early region promoter. Journal of Molecular and Applied Genetics 1:327–341
SpalholzB. A.,
LambertP. F.,
YeeC. L.,
HowleyP. M.1987; Bovine papillomavirus transcriptional regulation: localisation of the E2 responsive elements of the long control region. Journal of Virology 62:2125–2137
TsunokawaY.,
TakebeN.,
KasamatsuT.,
TeradaM.,
SugimuraT.1986; Transforming activity of human papillomavirus type 16 DNA sequences in a cervical cancer. Proceedings of the National Academy of SciencesU.S.A 83:2200–2203
WaulG. M.,
SternM.,
StarkG. R.1979; Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization using dextran sulfate. Proceedings of the National Academy of SciencesU.S.A 76:3683–3687
WatanabeS.,
YoshiikeK.1988; Transformation of rat 3Y1 cells by a deletion DNA of human papillomavirus type 16 molecularly cloned from genomic DNA of a cervical carcinoma. Journal of General Virology 69:1431–1435
WiglerM.,
SweetR.,
SimG. K.,
WoldB.,
PellicerA.,
LacyE.,
ManiatisT.,
SilversteinS.,
AxelR.1979; Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell 16:777–788
YasumotoS.,
BurkhardtA. L.,
DonigerJ.,
Di PaoloJ. A.1986; Human papillomavirus type 16 DNA induced malignant transformation of NIH 3T3 cells. Journal of Virology 57:572–577