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. 1989 Apr;86(7):2123–2127. doi: 10.1073/pnas.86.7.2123

Transactivation of both early and late simian virus 40 promoters by large tumor antigen does not require nuclear localization of the protein.

A G Wildeman 1
PMCID: PMC286863  PMID: 2538831

Abstract

The early gene product of simian virus 40, large tumor antigen (T antigen), is required for the onset of viral replication. This protein has also been reported to transactivate viral late gene expression, independently of replication. In this study I have used a vector that permits simultaneously a precise quantitation of simian virus 40 early and late promoter activity with a single nuclease S1 mapping probe. The results show that T antigen can activate the early promoter as well as the late promoter and that only on replicating templates does a shift occur in the ratio of late-to-early transcription. This simultaneous transactivation of early and late promoters occurs in human (HeLa) and monkey (CV-1) cells but does not occur in mouse embryonal carcinoma cells. It is seen with either wild-type T antigen or with a T antigen protein that carries a mutation in the nuclear localization signal. The mutant protein cannot bring about an early-to-late shift, consistent with its inability to support viral replication.

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Selected References

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

  1. Angel P., Imagawa M., Chiu R., Stein B., Imbra R. J., Rahmsdorf H. J., Jonat C., Herrlich P., Karin M. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor. Cell. 1987 Jun 19;49(6):729–739. doi: 10.1016/0092-8674(87)90611-8. [DOI] [PubMed] [Google Scholar]
  2. Barrera-Saldana H., Takahashi K., Vigneron M., Wildeman A., Davidson I., Chambon P. All six GC-motifs of the SV40 early upstream element contribute to promoter activity in vivo and in vitro. EMBO J. 1985 Dec 30;4(13B):3839–3849. doi: 10.1002/j.1460-2075.1985.tb04156.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berk A. J. Adenovirus promoters and E1A transactivation. Annu Rev Genet. 1986;20:45–79. doi: 10.1146/annurev.ge.20.120186.000401. [DOI] [PubMed] [Google Scholar]
  4. Borrelli E., Hen R., Chambon P. Adenovirus-2 E1A products repress enhancer-induced stimulation of transcription. Nature. 1984 Dec 13;312(5995):608–612. doi: 10.1038/312608a0. [DOI] [PubMed] [Google Scholar]
  5. Brady J., Bolen J. B., Radonovich M., Salzman N., Khoury G. Stimulation of simian virus 40 late gene expression by simian virus 40 tumor antigen. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2040–2044. doi: 10.1073/pnas.81.7.2040. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chalifour L. E., Wirak D. O., Hansen U., Wassarman P. M., DePamphilis M. L. cis- and trans-acting sequences required for expression of simian virus 40 genes in mouse oocytes. Genes Dev. 1987 Dec;1(10):1096–1106. doi: 10.1101/gad.1.10.1096. [DOI] [PubMed] [Google Scholar]
  7. Colledge W. H., Richardson W. D., Edge M. D., Smith A. E. Extensive mutagenesis of the nuclear location signal of simian virus 40 large-T antigen. Mol Cell Biol. 1986 Nov;6(11):4136–4139. doi: 10.1128/mcb.6.11.4136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DeCaprio J. A., Ludlow J. W., Figge J., Shew J. Y., Huang C. M., Lee W. H., Marsilio E., Paucha E., Livingston D. M. SV40 large tumor antigen forms a specific complex with the product of the retinoblastoma susceptibility gene. Cell. 1988 Jul 15;54(2):275–283. doi: 10.1016/0092-8674(88)90559-4. [DOI] [PubMed] [Google Scholar]
  9. DeLucia A. L., Lewton B. A., Tjian R., Tegtmeyer P. Topography of simian virus 40 A protein-DNA complexes: arrangement of pentanucleotide interaction sites at the origin of replication. J Virol. 1983 Apr;46(1):143–150. doi: 10.1128/jvi.46.1.143-150.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dean F. B., Bullock P., Murakami Y., Wobbe C. R., Weissbach L., Hurwitz J. Simian virus 40 (SV40) DNA replication: SV40 large T antigen unwinds DNA containing the SV40 origin of replication. Proc Natl Acad Sci U S A. 1987 Jan;84(1):16–20. doi: 10.1073/pnas.84.1.16. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ernoult-Lange M., Omilli F., May E. Contribution of different GC-motifs to the control of simian virus 40 late promoter activity. Nucleic Acids Res. 1987 Oct 26;15(20):8177–8193. doi: 10.1093/nar/15.20.8177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gallo G. J., Gilinger G., Alwine J. C. Simian virus 40 T antigen alters the binding characteristics of specific simian DNA-binding factors. Mol Cell Biol. 1988 Apr;8(4):1648–1656. doi: 10.1128/mcb.8.4.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gluzman Y., Frisque R. J., Sambrook J. Origin-defective mutants of SV40. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 1):293–300. doi: 10.1101/sqb.1980.044.01.033. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Green M., Loewenstein P. M., Pusztai R., Symington J. S. An adenovirus E1A protein domain activates transcription in vivo and in vitro in the absence of protein synthesis. Cell. 1988 Jun 17;53(6):921–926. doi: 10.1016/s0092-8674(88)90429-1. [DOI] [PubMed] [Google Scholar]
  16. Hansen U., Tenen D. G., Livingston D. M., Sharp P. A. T antigen repression of SV40 early transcription from two promoters. Cell. 1981 Dec;27(3 Pt 2):603–613. doi: 10.1016/0092-8674(81)90402-5. [DOI] [PubMed] [Google Scholar]
  17. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  18. Hoeffler W. K., Kovelman R., Roeder R. G. Activation of transcription factor IIIC by the adenovirus E1A protein. Cell. 1988 Jun 17;53(6):907–920. doi: 10.1016/s0092-8674(88)90409-6. [DOI] [PubMed] [Google Scholar]
  19. Imperiale M. J., Kao H. T., Feldman L. T., Nevins J. R., Strickland S. Common control of the heat shock gene and early adenovirus genes: evidence for a cellular E1A-like activity. Mol Cell Biol. 1984 May;4(5):867–874. doi: 10.1128/mcb.4.5.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Keller J. M., Alwine J. C. Activation of the SV40 late promoter: direct effects of T antigen in the absence of viral DNA replication. Cell. 1984 Feb;36(2):381–389. doi: 10.1016/0092-8674(84)90231-9. [DOI] [PubMed] [Google Scholar]
  21. Keller J. M., Alwine J. C. Analysis of an activatable promoter: sequences in the simian virus 40 late promoter required for T-antigen-mediated trans activation. Mol Cell Biol. 1985 Aug;5(8):1859–1869. doi: 10.1128/mcb.5.8.1859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kelly J. J., Munholland J. M., Wildeman A. G. Comeasurement of simian virus 40 early and late promoter activity in HeLa and 293 cells in the presence of T antigen. J Virol. 1989 Jan;63(1):383–391. doi: 10.1128/jvi.63.1.383-391.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kim C. H., Heath C., Bertuch A., Hansen U. Specific stimulation of simian virus 40 late transcription in vitro by a cellular factor binding the simian virus 40 21-base-pair repeat promoter element. Proc Natl Acad Sci U S A. 1987 Sep;84(17):6025–6029. doi: 10.1073/pnas.84.17.6025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kovesdi I., Reichel R., Nevins J. R. Role of an adenovirus E2 promoter binding factor in E1A-mediated coordinate gene control. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2180–2184. doi: 10.1073/pnas.84.8.2180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lebkowski J. S., Clancy S., Calos M. P. Simian virus 40 replication in adenovirus-transformed human cells antagonizes gene expression. Nature. 1985 Sep 12;317(6033):169–171. doi: 10.1038/317169a0. [DOI] [PubMed] [Google Scholar]
  26. Lee W., Mitchell P., Tjian R. Purified transcription factor AP-1 interacts with TPA-inducible enhancer elements. Cell. 1987 Jun 19;49(6):741–752. doi: 10.1016/0092-8674(87)90612-x. [DOI] [PubMed] [Google Scholar]
  27. Leong K., Brunet L., Berk A. J. Factors responsible for the higher transcriptional activity of extracts of adenovirus-infected cells fractionate with the TATA box transcription factor. Mol Cell Biol. 1988 Apr;8(4):1765–1774. doi: 10.1128/mcb.8.4.1765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lewis E. D., Manley J. L. Repression of simian virus 40 early transcription by viral DNA replication in human 293 cells. Nature. 1985 Sep 12;317(6033):172–175. doi: 10.1038/317172a0. [DOI] [PubMed] [Google Scholar]
  29. Lewton B. A., DeLucia A. L., Tegtmeyer P. Binding of simian virus 40 a protein to DNA with deletions at the origin of replication. J Virol. 1984 Jan;49(1):9–13. doi: 10.1128/jvi.49.1.9-13.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Loeken M. R., Khoury G., Brady J. Stimulation of the adenovirus E2 promoter by simian virus 40 T antigen or E1A occurs by different mechanisms. Mol Cell Biol. 1986 Jun;6(6):2020–2026. doi: 10.1128/mcb.6.6.2020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Loeken M., Bikel I., Livingston D. M., Brady J. trans-activation of RNA polymerase II and III promoters by SV40 small t antigen. Cell. 1988 Dec 23;55(6):1171–1177. doi: 10.1016/0092-8674(88)90261-9. [DOI] [PubMed] [Google Scholar]
  32. McBurney M. W., Rogers B. J. Isolation of male embryonal carcinoma cells and their chromosome replication patterns. Dev Biol. 1982 Feb;89(2):503–508. doi: 10.1016/0012-1606(82)90338-4. [DOI] [PubMed] [Google Scholar]
  33. Mitchell P. J., Wang C., Tjian R. Positive and negative regulation of transcription in vitro: enhancer-binding protein AP-2 is inhibited by SV40 T antigen. Cell. 1987 Sep 11;50(6):847–861. doi: 10.1016/0092-8674(87)90512-5. [DOI] [PubMed] [Google Scholar]
  34. Moran E. A region of SV40 large T antigen can substitute for a transforming domain of the adenovirus E1A products. Nature. 1988 Jul 14;334(6178):168–170. doi: 10.1038/334168a0. [DOI] [PubMed] [Google Scholar]
  35. Myers R. M., Rio D. C., Robbins A. K., Tjian R. SV40 gene expression is modulated by the cooperative binding of T antigen to DNA. Cell. 1981 Aug;25(2):373–384. doi: 10.1016/0092-8674(81)90056-8. [DOI] [PubMed] [Google Scholar]
  36. Pannuti A., Pascucci A., La Mantia G., Fisher-Fantuzzi L., Vesco C., Lania L. trans-activation of cellular and viral promoters by a transforming nonkaryophilic simian virus 40 large T antigen. J Virol. 1987 Apr;61(4):1296–1299. doi: 10.1128/jvi.61.4.1296-1299.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Reed S. I., Stark G. R., Alwine J. C. Autoregulation of simian virus 40 gene A by T antigen. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3083–3087. doi: 10.1073/pnas.73.9.3083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rio D., Robbins A., Myers R., Tjian R. Regulation of simian virus 40 early transcription in vitro by a purified tumor antigen. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5706–5710. doi: 10.1073/pnas.77.10.5706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Scher B. M., Dean D. H., Garro A. J. Fragmentation of Bacillus bacteriophage phi105 DNA by complementary single-stranded DNA in the cohesive ends of the molecule. J Virol. 1977 Aug;23(2):377–383. doi: 10.1128/jvi.23.2.377-383.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tegtmeyer P. Function of simian virus 40 gene A in transforming infection. J Virol. 1975 Mar;15(3):613–618. doi: 10.1128/jvi.15.3.613-618.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tegtmeyer P. Simian virus 40 deoxyribonucleic acid synthesis: the viral replicon. J Virol. 1972 Oct;10(4):591–598. doi: 10.1128/jvi.10.4.591-598.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Velcich A., Ziff E. Adenovirus E1a proteins repress transcription from the SV40 early promoter. Cell. 1985 Mar;40(3):705–716. doi: 10.1016/0092-8674(85)90219-3. [DOI] [PubMed] [Google Scholar]
  43. Whyte P., Buchkovich K. J., Horowitz J. M., Friend S. H., Raybuck M., Weinberg R. A., Harlow E. Association between an oncogene and an anti-oncogene: the adenovirus E1A proteins bind to the retinoblastoma gene product. Nature. 1988 Jul 14;334(6178):124–129. doi: 10.1038/334124a0. [DOI] [PubMed] [Google Scholar]
  44. Zenke M., Grundström T., Matthes H., Wintzerith M., Schatz C., Wildeman A., Chambon P. Multiple sequence motifs are involved in SV40 enhancer function. EMBO J. 1986 Feb;5(2):387–397. doi: 10.1002/j.1460-2075.1986.tb04224.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. de Villiers J., Schaffner W. A small segment of polyoma virus DNA enhances the expression of a cloned beta-globin gene over a distance of 1400 base pairs. Nucleic Acids Res. 1981 Dec 11;9(23):6251–6264. doi: 10.1093/nar/9.23.6251. [DOI] [PMC free article] [PubMed] [Google Scholar]

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