Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1983 Mar;3(3):484–489. doi: 10.1128/mcb.3.3.484

Simian virus 40 deletion mutants that transform with reduced efficiency.

L Sompayrac, K J Danna
PMCID: PMC368559  PMID: 6302476

Abstract

We have constructed two simian virus 40 (SV40) early-region deletion mutants that lack a significant portion of the sequences normally used to encode the SV40 large tumor antigen. Despite these deletions, the mutants were able to transform mouse cells in a focus assay, although with a frequency that was drastically reduced relative to wild-type SV40. Cell lines expanded from the mutant-transformed foci contained integrated mutant DNA, expressed an SV40 tumor antigen (small-t), and exhibited a range of transformed phenotypes, which included the ability to grow while suspended in soft agar. We also present evidence that these mutants are defective for abortive transformation in an assay that tested the transient loss of anchorage dependence. Their ability to stably transform, contrasted with their inability to abortively transform at detectable levels, raises the possibility that the mechanism by which these mutants transform may be different from that of wild-type SV40.

Full text

PDF
486

Images in this article

485Image
on p.485
486Image
on p.486

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Clayton C. E., Murphy D., Lovett M., Rigby P. W. A fragment of the SV40 large T-antigen gene transforms. Nature. 1982 Sep 2;299(5878):59–61. doi: 10.1038/299059a0. [DOI] [PubMed] [Google Scholar]
  2. Collett M. S., Erikson R. L. Protein kinase activity associated with the avian sarcoma virus src gene product. Proc Natl Acad Sci U S A. 1978 Apr;75(4):2021–2024. doi: 10.1073/pnas.75.4.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  4. Gross-Bellard M., Oudet P., Chambon P. Isolation of high-molecular-weight DNA from mammalian cells. Eur J Biochem. 1973 Jul 2;36(1):32–38. doi: 10.1111/j.1432-1033.1973.tb02881.x. [DOI] [PubMed] [Google Scholar]
  5. Hayward W. S., Neel B. G., Astrin S. M. Activation of a cellular onc gene by promoter insertion in ALV-induced lymphoid leukosis. Nature. 1981 Apr 9;290(5806):475–480. doi: 10.1038/290475a0. [DOI] [PubMed] [Google Scholar]
  6. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  7. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Reznikoff C. A., Brankow D. W., Heidelberger C. Establishment and characterization of a cloned line of C3H mouse embryo cells sensitive to postconfluence inhibition of division. Cancer Res. 1973 Dec;33(12):3231–3238. [PubMed] [Google Scholar]
  9. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  10. Risser R., Pollack R. A nonselective analysis of SV40 transformation of mouse 3T3 cells. Virology. 1974 Jun;59(2):477–489. doi: 10.1016/0042-6822(74)90457-7. [DOI] [PubMed] [Google Scholar]
  11. Sompayrac L. M., Danna K. J. Efficient infection of monkey cells with DNA of simian virus 40. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7575–7578. doi: 10.1073/pnas.78.12.7575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sompayrac L. M., Danna K. J. Isolation and characterization of simian virus 40 early region deletion mutants. J Virol. 1982 Jul;43(1):328–331. doi: 10.1128/jvi.43.1.328-331.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Sompayrac L. M., Gurney E. G., Danna K. J. Stabilization of the 53,000-dalton nonviral tumor antigen is not required for transformation by simian virus 40. Mol Cell Biol. 1983 Feb;3(2):290–296. doi: 10.1128/mcb.3.2.290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  15. Stoker M. Abortive transformation by polyoma virus. Nature. 1968 Apr 20;218(5138):234–238. doi: 10.1038/218234a0. [DOI] [PubMed] [Google Scholar]
  16. Stoker M., Dulbecco R. Abortive transformation by the Tsa mutant of polyoma virus. Nature. 1969 Jul 26;223(5204):397–398. doi: 10.1038/223397a0. [DOI] [PubMed] [Google Scholar]
  17. Wigler M., Pellicer A., Silverstein S., Axel R., Urlaub G., Chasin L. DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1373–1376. doi: 10.1073/pnas.76.3.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES