Transformation mediated by the SV40 T antigens: separation of the overlapping SV40 early genes with a retroviral vector

Abstract

A murine retroviral vector has been used to separate physically the overlapping genes encoded by SV40. This minimal retroviral vector contains LTRs and other cis-acting signals required for infectious RNA virus propagation. We placed the SV40 early region within this DNA and after transfection of cells producing helper Moloney murine leukemia virus, SV40 retroviruses (MV40) could be rescued. Cytoplasmic spliced large T and small t transcripts, as well as unspliced transcripts, are packaged into virions with equal efficiency. Pure SV40 large T retroviruses can be cloned from these heterogeneous virus stocks by secondary transformation of rodent cells. The large T retrovirus stocks morphologically transform primary or established mouse and rat lines with high efficiency. There is little difference in transformation either by agar assay or focus formation between retroviruses carrying both SV40 genes or large T alone. We present quantitative data that demonstrate that abortive transformation of rodent cells by SV40, transient expression of the transformed phenotype after infection, is not manifested by MV40. Thus abortive transformation is not the result of a weakly dominant transforming gene, but rather of the normally inefficient mode of integration and early gene expression of SV40 upon infection of rodent cells.