Separation of immortalization from tumor induction with polyoma large T mutants that fail to bind the retinoblastoma gene product.

The mouse polyomavirus encodes a tumor-suppressor gene inactivator in its large T protein and a proto-oncogene activator in its middle T protein. We have used site-directed mutagenesis to selectively inactivate the former function without affecting the latter. Two mutant viruses were constructed to encode altered large T proteins that fail to bind the retinoblastoma tumor-suppressor gene product pRB, along with normal small and middle T proteins. The pRB-binding mutants proved to be defective in immortalization of primary rat embryo fibroblasts by a variety of tests. Yet they proved capable of transforming both primary and established fibroblasts in culture. Most importantly, the inability of these mutants to bind pRB had little effect on their ability to induce tumors in mice. We conclude that induction of multiple tumor types in this system does not depend on large T-pRB interactions but rather on middle T-dependent pathways. In addition, the ability of this virus to immortalize cells in culture is not essential to its ability to induce tumors in the animal.