The E5 gene of bovine papillomavirus type 1 is sufficient for complete oncogenic transformation of mouse fibroblasts

Abstract

The transforming 69% fragment of the bovine papillomavirus type-1 genome was inserted into a retrovirus vector, which also expresses G418 resistance, and the resulting construct was used for transfection of psi 2 cells. C127 cells infected with virus-containing medium from G418 resistant psi 2 clones were selected for G418 resistance and/or transformation. G418 resistant cells contained invariably a 4.4 kb provirus. The transformed cells, in contrast, contained either a 2.8 kb or a 5.4 kb provirus. Cells containing the 2.8 or the 5.4 kb proviruses were fully transformed according to several criteria: they had a transformed morphology, formed colonies in soft agar, contained disarranged F-actin cables and induced tumors when injected subcutaneously into nude mice. A molecular analysis of the 2.8 kb provirus showed that it contained only one complete BPV-1 gene, namely E5. Cells transformed by the 2.8 kb provirus contained colinear RNAs as well as subgenomic mRNAs, composed of a retrovirus leader sequence connected to an exon starting at nucleotide 3605 in the BPV-1 sequence. The only papillomavirus protein that can be expressed from these mRNAs is the E5 protein. The results suggest that the 44 amino acid long membrane protein which is encoded by the E5 gene of BPV-1, confers a fully transformed phenotype to immortalized mouse cells in the absence of other viral gene products or papillomavirus regulatory elements.