“Hit and run” transformation of mouse c127 cells by Bovine Papillomavirus type 4: The viral DNA is required for the initiation but not for maintenance of the transformed phenotype

Abstract

Morphological transformation of C127 mouse fibroblasts by bovine papillomavirus type 4 (BPV-4) DNA depends on additional factors, including cell density, the presence of TPA, the concentration of fetal calf serum, and the physical state of the input DNA. Low cell density or the presence of TPA allows the achievement of full transformation, suggesting that disturbance of cell-to-cell contact may be necessary for the expression of the malignant phenotype. TPA also induces a burst of viral DNA synthesis and transcription. The frequency of focus formation is fivefold higher in the presence of high serum than in low serum, indicating that growth factors are required during the early stages of transformation. Although the same number of foci is obtained with either circular or linear DNA, the cells transfected with linear DNA are at a more advanced stage of transformation, being capable of anchorage-independent growth and tumorigenic in nude mice; only 5 out of 21 of these cell lines contain viral DNA in integrated and rearranged forms. In contrast cells transfected with circular DNA are anchorage-dependent but tumorigenic in mice, and do not contain any viral DNA. The linear DNA has an interrupted E1 ORF and fully transformed cells are obtained also with a subgenomic viral DNA fragment containing part of the Ll ORF, the complete E6, E7, and E8 ORFs, and the truncated E1 ORF under the control of the BPV-4 regulatory region; viral DNA is not maintained in any of these cell lines. In total, only 15% of the transformed cell lines contains viral DNA. These results are discussed in the light of the “hit and run” model of cell transformation.