Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1972 Jan;69(1):152–156. doi: 10.1073/pnas.69.1.152

Detection and Quantitation of Simian Virus 40 Genetic Material in Abortively Transformed BALB/3T3 Clones

Helene S Smith *,†, Lawrence D Gelb 2, Malcolm A Martin 2
PMCID: PMC427565  PMID: 4333038

Abstract

Infection with simian virus 40 is known to induce many cells to synthesize DNA and to divide in a medium lacking serum protein growth factor(s) that is essential for growth of uninfected cells (factor-free medium). Cells infected under these conditions then go through several rounds of division, since colonies containing more than 100 cells are formed. Many of these colonies are abortively transformed since, upon subsequent passage of the cells in standard medium, they can no longer grow in factor-free medium and show no other properties of viral transformation. We have examined these abortively transformed cells for the presence of simian virus 40 DNA sequences. Of the three clones tested, two were found to contain viral genetic material despite the fact that they were phenotypically normal.

The number of simian virus 40 genome equivalents present was determined by measurement of DNA reassociation kinetics on hydroxyapatite. Two of the abortively transformed lines contained approximately five viral genome equivalents per diploid cell, while the DNA from a third abortive transformant was indistinguishable from that of uninfected BALB/3T3 cells. A standard simian virus 40 transformant, isolated under similar conditions, contained two copies of the viral genome per cell. The abortive transformants also appear to contain the entire viral genome rather than multiple partial copies. Subclones of one abortively transformed line containing five copies per cell had 2.7-10 copies of viral genetic material per diploid cell.

Keywords: mice, diploid cells, virus equivalents

Full text

PDF
152

Selected References

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

  1. AUB J. C., TIESLAU C., LANKESTER A. REACTIONS OF NORMAL AND TUMOR CELL SURFACES TO ENZYMES. I. WHEAT-GERM LIPASE AND ASSOCIATED MUCOPOLYSACCHARIDES. Proc Natl Acad Sci U S A. 1963 Oct;50:613–619. doi: 10.1073/pnas.50.4.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aaronson S. A., Todaro G. J. Development of 3T3-like lines from Balb-c mouse embryo cultures: transformation susceptibility to SV40. J Cell Physiol. 1968 Oct;72(2):141–148. doi: 10.1002/jcp.1040720208. [DOI] [PubMed] [Google Scholar]
  3. BLACK P. H., ROWE W. P., TURNER H. C., HUEBNER R. J. A SPECIFIC COMPLEMENT-FIXING ANTIGEN PRESENT IN SV40 TUMOR AND TRANSFORMED CELLS. Proc Natl Acad Sci U S A. 1963 Dec;50:1148–1156. doi: 10.1073/pnas.50.6.1148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Britten R. J., Kohne D. E. Repeated sequences in DNA. Hundreds of thousands of copies of DNA sequences have been incorporated into the genomes of higher organisms. Science. 1968 Aug 9;161(3841):529–540. doi: 10.1126/science.161.3841.529. [DOI] [PubMed] [Google Scholar]
  5. Eckhart W. Cell transformation by polyoma virus and SV40. Nature. 1969 Dec 13;224(5224):1069–1071. doi: 10.1038/2241069a0. [DOI] [PubMed] [Google Scholar]
  6. Gelb L. D., Kohne D. E., Martin M. A. Quantitation of Simian virus 40 sequences in African green monkey, mouse and virus-transformed cell genomes. J Mol Biol. 1971 Apr 14;57(1):129–145. doi: 10.1016/0022-2836(71)90123-9. [DOI] [PubMed] [Google Scholar]
  7. Gerber P. Studies on the transfer of subviral infectivity from SV40-induced hamster tumor cells to indicator cells. Virology. 1966 Apr;28(4):501–509. doi: 10.1016/0042-6822(66)90234-0. [DOI] [PubMed] [Google Scholar]
  8. Gershon D., Sachs L., Winocour E. The induction of cellular DNA synthesis by simian virus 40 in contact-inhibited and in x-irradiated cells. Proc Natl Acad Sci U S A. 1966 Sep;56(3):918–925. doi: 10.1073/pnas.56.3.918. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. HABEL K., EDDY B. E. Specificity of resistance to tumor challenge of polyoma and SV 40 virus-immune hamsters. Proc Soc Exp Biol Med. 1963 May;113:1–4. doi: 10.3181/00379727-113-28259. [DOI] [PubMed] [Google Scholar]
  10. Henry P., Black P. H., Oxman M. N., Weissman S. M. Stimulation of DNA synthesis in mouse cell line 3T3 by Simian virus 40. Proc Natl Acad Sci U S A. 1966 Oct;56(4):1170–1176. doi: 10.1073/pnas.56.4.1170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Huebner R. J., Todaro G. J. Oncogenes of RNA tumor viruses as determinants of cancer. Proc Natl Acad Sci U S A. 1969 Nov;64(3):1087–1094. doi: 10.1073/pnas.64.3.1087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Inbar M., Sachs L. Interaction of the carbohydrate-binding protein concanavalin A with normal and transformed cells. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1418–1425. doi: 10.1073/pnas.63.4.1418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jainchill J. L., Todaro G. J. Stimulation of cell growth in vitro by serum with and without growth factor. Relation to contact inhibition and viral transformation. Exp Cell Res. 1970 Jan;59(1):137–146. doi: 10.1016/0014-4827(70)90632-4. [DOI] [PubMed] [Google Scholar]
  14. Koprowski H., Jensen F. C., Steplewski Z. Activation of production of infectious tumor virus SV40 in heterokaryon cultures. Proc Natl Acad Sci U S A. 1967 Jul;58(1):127–133. doi: 10.1073/pnas.58.1.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Maass G., Werchau H., Brandner G., Haas R. Incorporation of iododeoxyuridine into cellular deoxyribonucleic acid synthesized after infection with simian virus 40. J Virol. 1968 Jul;2(7):723–727. doi: 10.1128/jvi.2.7.723-727.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pollack R. E., Green H., Todaro G. J. Growth control in cultured cells: selection of sublines with increased sensitivity to contact inhibition and decreased tumor-producing ability. Proc Natl Acad Sci U S A. 1968 May;60(1):126–133. doi: 10.1073/pnas.60.1.126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sambrook J., Westphal H., Srinivasan P. R., Dulbecco R. The integrated state of viral DNA in SV40-transformed cells. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1288–1295. doi: 10.1073/pnas.60.4.1288. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Smith H. S., Scher C. D., Todaro G. J. Induction of cell division in medium lacking serum growth factor by SV40. Virology. 1971 May;44(2):359–370. doi: 10.1016/0042-6822(71)90267-4. [DOI] [PubMed] [Google Scholar]
  19. Stoker M., O'Neill C., Berryman S., Waxman V. Anchorage and growth regulation in normal and virus-transformed cells. Int J Cancer. 1968 Sep 15;3(5):683–693. doi: 10.1002/ijc.2910030517. [DOI] [PubMed] [Google Scholar]
  20. Watkins J. F., Dulbecco R. Production of SV40 virus in heterokaryons of transformed and susceptible cells. Proc Natl Acad Sci U S A. 1967 Oct;58(4):1396–1403. doi: 10.1073/pnas.58.4.1396. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES