Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 1997 Aug;146(4):1417–1428. doi: 10.1093/genetics/146.4.1417

Induction of Duplication Reversion in Human Fibroblasts, by Wild-Type and Mutated Sv40 T Antigen, Covaries with the Ability to Induce Host DNA Synthesis

M A Shammas 1, S J Xia 1, RJS Reis 1
PMCID: PMC1208085  PMID: 9258684

Abstract

Intrachromosomal homologous recombination, manifest as reversion of a 14-kbp duplication in the hypoxanthine phosphoribosyl transferase (HPRT) gene, is elevated in human cells either stably transformed or transiently transfected by the SV40 (simian virus 40) large T antigen gene. Following introduction of wild-type SV40, or any of several T-antigen point mutations in a constant SV40 background, we observed a strong correlation between the stimulation of chromosomal recombination and induction of host-cell DNA synthesis. Moreover, inhibitors of DNA replication (aphidicolin and hydroxyurea) suppress SV40-induced homologous recombination to the extent that they suppress DNA synthesis. Stable integration of plasmids encoding T antigen also augments homologous recombination, which is suppressed by aphidicolin. We infer that the mechanism by which T antigen stimulates homologous recombination in human fibroblasts involves DNA replicative synthesis.

Full Text

The Full Text of this article is available as a PDF (3.8 MB).

Selected References

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

  1. Baumstark-Khan C. Effect of aphidicolin on DNA synthesis, PLD-recovery and DNA repair of human diploid fibroblasts. Int J Radiat Biol. 1992 Feb;61(2):191–197. doi: 10.1080/09553009214550811. [DOI] [PubMed] [Google Scholar]
  2. Borowiec J. A., Hurwitz J. Localized melting and structural changes in the SV40 origin of replication induced by T-antigen. EMBO J. 1988 Oct;7(10):3149–3158. doi: 10.1002/j.1460-2075.1988.tb03182.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brinster R. L., Chen H. Y., Messing A., van Dyke T., Levine A. J., Palmiter R. D. Transgenic mice harboring SV40 T-antigen genes develop characteristic brain tumors. Cell. 1984 Jun;37(2):367–379. doi: 10.1016/0092-8674(84)90367-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chou J. Y., Martin R. G. DNA infectivity and the induction of host DNA synthesis with temperature-sensitive mutants of simian virus 40. J Virol. 1975 Jan;15(1):145–150. doi: 10.1128/jvi.15.1.145-150.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen A., Clark A. J. Synthesis of linear plasmid multimers in Escherichia coli K-12. J Bacteriol. 1986 Jul;167(1):327–335. doi: 10.1128/jb.167.1.327-335.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dickmanns A., Zeitvogel A., Simmersbach F., Weber R., Arthur A. K., Dehde S., Wildeman A. G., Fanning E. The kinetics of simian virus 40-induced progression of quiescent cells into S phase depend on four independent functions of large T antigen. J Virol. 1994 Sep;68(9):5496–5508. doi: 10.1128/jvi.68.9.5496-5508.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dodson M., Dean F. B., Bullock P., Echols H., Hurwitz J. Unwinding of duplex DNA from the SV40 origin of replication by T antigen. Science. 1987 Nov 13;238(4829):964–967. doi: 10.1126/science.2823389. [DOI] [PubMed] [Google Scholar]
  8. Drews R. E., Chan V. T., Schnipper L. E. Oncogenes result in genomic alterations that activate a transcriptionally silent, dominantly selectable reporter gene (neo). Mol Cell Biol. 1992 Jan;12(1):198–206. doi: 10.1128/mcb.12.1.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Drize O. B., Sokova O. I., Nikashina E. B., Shliankevich M. A., Shapot V. S. Vozmozhnaia rol' T-antigena v induktsii khromosomnykh aberratsii v kletkakh, transformirovannykh virusom SV40. Tsitologiia. 1985 Jan;27(1):76–82. [PubMed] [Google Scholar]
  10. Elford H. L. Effect of hydroxyurea on ribonucleotide reductase. Biochem Biophys Res Commun. 1968 Oct 10;33(1):129–135. doi: 10.1016/0006-291x(68)90266-0. [DOI] [PubMed] [Google Scholar]
  11. Esposito M. S. Evidence that spontaneous mitotic recombination occurs at the two-strand stage. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4436–4440. doi: 10.1073/pnas.75.9.4436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fanning E., Knippers R. Structure and function of simian virus 40 large tumor antigen. Annu Rev Biochem. 1992;61:55–85. doi: 10.1146/annurev.bi.61.070192.000415. [DOI] [PubMed] [Google Scholar]
  13. Finn G. K., Kurz B. W., Cheng R. Z., Shmookler Reis R. J. Homologous plasmid recombination is elevated in immortally transformed cells. Mol Cell Biol. 1989 Sep;9(9):4009–4017. doi: 10.1128/mcb.9.9.4009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gannon J. V., Lane D. P. Interactions between SV40 T antigen and DNA polymerase alpha. New Biol. 1990 Jan;2(1):84–92. [PubMed] [Google Scholar]
  15. Goldstein S., Fordis C. M., Howard B. H. Enhanced transfection efficiency and improved cell survival after electroporation of G2/M-synchronized cells and treatment with sodium butyrate. Nucleic Acids Res. 1989 May 25;17(10):3959–3971. doi: 10.1093/nar/17.10.3959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gorman S. D., Cristofalo V. J. Reinitiation of cellular DNA synthesis in BrdU-selected nondividing senescent WI-38 cells by simian virus 40 infection. J Cell Physiol. 1985 Oct;125(1):122–126. doi: 10.1002/jcp.1041250116. [DOI] [PubMed] [Google Scholar]
  17. Gurney T., Jr, Gurney E. G. Spontaneous rearrangement of integrated simian virus 40 DNA in nine transformed rodent cell lines. J Virol. 1989 Jan;63(1):165–174. doi: 10.1128/jvi.63.1.165-174.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hanahan D. Heritable formation of pancreatic beta-cell tumours in transgenic mice expressing recombinant insulin/simian virus 40 oncogenes. Nature. 1985 May 9;315(6015):115–122. doi: 10.1038/315115a0. [DOI] [PubMed] [Google Scholar]
  19. Hara E., Uzman J. A., Dimri G. P., Nehlin J. O., Testori A., Campisi J. The helix-loop-helix protein Id-1 and a retinoblastoma protein binding mutant of SV40 T antigen synergize to reactivate DNA synthesis in senescent human fibroblasts. Dev Genet. 1996;18(2):161–172. doi: 10.1002/(SICI)1520-6408(1996)18:2<161::AID-DVG9>3.0.CO;2-7. [DOI] [PubMed] [Google Scholar]
  20. Kawasaki I., Bae Y. S., Eki T., Kim Y., Ikeda H. Homologous recombination of monkey alpha-satellite repeats in an in vitro simian virus 40 replication system: possible association of recombination with DNA replication. Mol Cell Biol. 1994 Jun;14(6):4173–4182. doi: 10.1128/mcb.14.6.4173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kogoma T. RNase H-defective mutants of Escherichia coli. J Bacteriol. 1986 May;166(2):361–363. doi: 10.1128/jb.166.2.361-363.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kogoma T. Recombination by replication. Cell. 1996 May 31;85(5):625–627. doi: 10.1016/s0092-8674(00)81229-5. [DOI] [PubMed] [Google Scholar]
  23. Land H., Parada L. F., Weinberg R. A. Cellular oncogenes and multistep carcinogenesis. Science. 1983 Nov 18;222(4625):771–778. doi: 10.1126/science.6356358. [DOI] [PubMed] [Google Scholar]
  24. Lawrence C. W. Effect of fractionated doses of radiation on recombination. Heredity (Edinb) 1968 Feb;23(1):143–146. doi: 10.1038/hdy.1968.14. [DOI] [PubMed] [Google Scholar]
  25. Loeber G., Stenger J. E., Ray S., Parsons R. E., Anderson M. E., Tegtmeyer P. The zinc finger region of simian virus 40 large T antigen is needed for hexamer assembly and origin melting. J Virol. 1991 Jun;65(6):3167–3174. doi: 10.1128/jvi.65.6.3167-3174.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Loeber G., Tevethia M. J., Schwedes J. F., Tegtmeyer P. Temperature-sensitive mutants identify crucial structural regions of simian virus 40 large T antigen. J Virol. 1989 Oct;63(10):4426–4430. doi: 10.1128/jvi.63.10.4426-4430.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Melendy T., Stillman B. An interaction between replication protein A and SV40 T antigen appears essential for primosome assembly during SV40 DNA replication. J Biol Chem. 1993 Feb 15;268(5):3389–3395. [PubMed] [Google Scholar]
  28. Milner J., Gamble J. The SV40 large T-p53 complex: evidence for the presence of two immunologically distinct forms of p53. Virology. 1985 Nov;147(1):206–209. doi: 10.1016/0042-6822(85)90240-5. [DOI] [PubMed] [Google Scholar]
  29. Montenarh M., Kohler M., Henning R. Oligomerization of simian virus 40 large T antigen is not necessarily repressed by temperature-sensitive A gene lesions. J Virol. 1984 Mar;49(3):658–664. doi: 10.1128/jvi.49.3.658-664.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Morgan A. R., Severini A. Interconversion of replication and recombination structures: implications for terminal repeats and concatemers. J Theor Biol. 1990 May 22;144(2):195–202. doi: 10.1016/s0022-5193(05)80318-2. [DOI] [PubMed] [Google Scholar]
  31. Murakami Y., Hurwitz J. DNA polymerase alpha stimulates the ATP-dependent binding of simian virus tumor T antigen to the SV40 origin of replication. J Biol Chem. 1993 May 25;268(15):11018–11027. [PubMed] [Google Scholar]
  32. Oshima J., Steinmann K. E., Campisi J., Schlegel R. Modulation of cell growth, p34cdc2 and cyclin A levels by SV-40 large T antigen. Oncogene. 1993 Nov;8(11):2987–2993. [PubMed] [Google Scholar]
  33. Parsons R., Anderson M. E., Tegtmeyer P. Three domains in the simian virus 40 core origin orchestrate the binding, melting, and DNA helicase activities of T antigen. J Virol. 1990 Feb;64(2):509–518. doi: 10.1128/jvi.64.2.509-518.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Pintel D., Bouck N., di Mayorca G. Separation of lytic and transforming functions of the simian virus 40 A region: two mutants which are temperature sensitive for lytic functions have opposite effects on transformation. J Virol. 1981 May;38(2):518–528. doi: 10.1128/jvi.38.2.518-528.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Pistillo J. M., Vishwanatha J. K. Interaction of simian virus 40 large T-antigen with cellular DNA polymerase alpha: studies with various T-antigen mutants of SV40. Arch Virol. 1991;118(1-2):113–125. doi: 10.1007/BF01311307. [DOI] [PubMed] [Google Scholar]
  36. Radman M., Jeggo P., Wagner R. Chromosomal rearrangement and carcinogenesis. Mutat Res. 1982 May;98(3):249–264. doi: 10.1016/0165-1110(82)90035-5. [DOI] [PubMed] [Google Scholar]
  37. Ray F. A., Peabody D. S., Cooper J. L., Cram L. S., Kraemer P. M. SV40 T antigen alone drives karyotype instability that precedes neoplastic transformation of human diploid fibroblasts. J Cell Biochem. 1990 Jan;42(1):13–31. doi: 10.1002/jcb.240420103. [DOI] [PubMed] [Google Scholar]
  38. Ray S., Anderson M. E., Loeber G., McVey D., Tegtmeyer P. Functional characterization of temperature-sensitive mutants of simian virus 40 large T antigen. J Virol. 1992 Nov;66(11):6509–6516. doi: 10.1128/jvi.66.11.6509-6516.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Ray S., Anderson M. E., Tegtmeyer P. Differential interaction of temperature-sensitive simian virus 40 T antigens with tumor suppressors pRb and p53. J Virol. 1996 Oct;70(10):7224–7227. doi: 10.1128/jvi.70.10.7224-7227.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Razin S., Pfendt E. A., Matsumura T., Hayflick L. Comparison by autoradiography of macromolecular biosynthesis in "young" and "old" human diploid fibroblast cultures. A brief note. Mech Ageing Dev. 1977 Sep-Oct;6(5):379–384. doi: 10.1016/0047-6374(77)90039-2. [DOI] [PubMed] [Google Scholar]
  41. Rodarte-Ramón U. S. Radiation-induced recombination in Saccharomyces: the genetic control of recombination in mitosis and meiosis. Radiat Res. 1972 Jan;49(1):148–154. [PubMed] [Google Scholar]
  42. Subramani S., Rubnitz J. Recombination events after transient infection and stable integration of DNA into mouse cells. Mol Cell Biol. 1985 Apr;5(4):659–666. doi: 10.1128/mcb.5.4.659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Swann P. F., Waters T. R., Moulton D. C., Xu Y. Z., Zheng Q., Edwards M., Mace R. Role of postreplicative DNA mismatch repair in the cytotoxic action of thioguanine. Science. 1996 Aug 23;273(5278):1109–1111. doi: 10.1126/science.273.5278.1109. [DOI] [PubMed] [Google Scholar]
  44. Viret J. F., Alonso J. C. Generation of linear multigenome-length plasmid molecules in Bacillus subtilis. Nucleic Acids Res. 1987 Aug 25;15(16):6349–6367. doi: 10.1093/nar/15.16.6349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Wilson V. G., Tevethia M. J., Lewton B. A., Tegtmeyer P. DNA binding properties of simian virus 40 temperature-sensitive A proteins. J Virol. 1982 Nov;44(2):458–466. doi: 10.1128/jvi.44.2.458-466.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wong E. A., Capecchi M. R. Homologous recombination between coinjected DNA sequences peaks in early to mid-S phase. Mol Cell Biol. 1987 Jun;7(6):2294–2295. doi: 10.1128/mcb.7.6.2294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wright W. E., Pereira-Smith O. M., Shay J. W. Reversible cellular senescence: implications for immortalization of normal human diploid fibroblasts. Mol Cell Biol. 1989 Jul;9(7):3088–3092. doi: 10.1128/mcb.9.7.3088. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Xiong Y., Zhang H., Beach D. Subunit rearrangement of the cyclin-dependent kinases is associated with cellular transformation. Genes Dev. 1993 Aug;7(8):1572–1583. doi: 10.1101/gad.7.8.1572. [DOI] [PubMed] [Google Scholar]
  49. Yang T. P., Stout J. T., Konecki D. S., Patel P. I., Alford R. L., Caskey C. T. Spontaneous reversion of novel Lesch-Nyhan mutation by HPRT gene rearrangement. Somat Cell Mol Genet. 1988 May;14(3):293–303. doi: 10.1007/BF01534590. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES