Expression of the large T protein of polyoma virus promotes the establishment in culture of "normal" rodent fibroblast cell lines.
- 1 July 1983
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 80 (14), 4354-4358
- https://doi.org/10.1073/pnas.80.14.4354
Abstract
Transfer into mouse and rat embryo fibroblasts in primary culture of cloned polyoma virus genes encoding only the large T protein led to the establishment of flat colonies in sparse subcultures at a frequency equal to that of transformation by wild-type virus. cell lines could be derived from such colonies and maintained in culture for large numbers of generations without entering crisis. They exhibited a normal phenotype by the criteria of growth on plastic to a low saturation density and of anchorage dependency. They required a lower serum concentration for growth than spontaneously established [mouse] 3T3 cells. Similar results were obtained after transfer of recombinant DNA molecules encoding only the amino-terminal 40% of the large T protein, suggesting that this immortalization function corresponds to the activity of an amino-terminal domain of the protein. Immunoprecipitation analysis of T antigens in cell lines established after transfer of the full-size and of the truncated large T genes demonstrated the expression of the full-size large-T-protein and of a MW 40,000 antigen expressed from the amino-terminal part of the gene, respectively. After transfer of a large T only plasmid that carries a tsa mutation, cell lines were established at 33.degree. C with the same efficiency as with the wild-type large T gene, but their growth was arrested after a shift to 40.degree. C with a progressive loss in cell viability. This result indicates a continuous requirement for a large T function in the maintenance of immortality.This publication has 25 references indexed in Scilit:
- A fragment of the SV40 large T-antigen gene transformsNature, 1982
- High frequency of gene transfer after fusion between bacteria and eukaryotic cellsNature, 1982
- Coding potential and regulatory signals of the polyoma virus genomeNature, 1980
- Untransformed rat cells containing free and integrated dna of a polyoma nontransforming (Hr-t) mutantCell, 1979
- The roles of the simian virus 40 tumor antigens in transformation of chinese hamster lung cellsCell, 1979
- Factors affecting the frequency of transformation of rat embryo cells by simian virus 40Virology, 1979
- Tumor antigens induced by nontransforming mutants of polyoma virusCell, 1978
- A new technique for the assay of infectivity of human adenovirus 5 DNAVirology, 1973
- Isolation of temperature-sensitive mutants of polyoma virusVirology, 1965
- QUANTITATIVE STUDIES OF THE GROWTH OF MOUSE EMBRYO CELLS IN CULTURE AND THEIR DEVELOPMENT INTO ESTABLISHED LINESThe Journal of cell biology, 1963