Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1991 Dec;65(12):6553–6561. doi: 10.1128/jvi.65.12.6553-6561.1991

Simian virus 40 small-t does not transactivate RNA polymerase II promoters in virus infections.

P Rajan 1, V Dhamankar 1, K Rundell 1, B Thimmapaya 1
PMCID: PMC250710  PMID: 1658360

Abstract

Transcriptional stimulatory properties of virus-encoded transactivators appear to be critical for viral gene expression and may be linked to cellular transformation in certain cases. Recently, the simian virus 40 (SV40) 17-kDa small-t antigen was shown to stimulate transcription of polymerase II and III genes in transient transfection assays. In experiments performed in our laboratory, two of the polymerase II promoters of the adenovirus genome, namely, the EII-early and EIII promoters, were transactivation, we examined the transient transfection assays. To further elucidate the mechanism of this transactivation, we examined the ability of small-t to transactivate the adenovirus type 5 EII-early and EIII promoters in CV-1 cells under conditions in which the small-t gene or the reporter genes were introduced into the cells through transfection and other routes. In one approach, we used established CV-1 cell lines which constitutively express the small-t gene, and study of the EII-early promoter was afforded by infection of an EIA-negative adenovirus type 5 variant. For the second approach, a recombinant adenovirus was constructed in which small-t was expressed from a replication origin-negative SV40 early promoter in the EIA region of an adenovirus vector (Ad-SV-t). The effect of small-t on adenovirus EII-early and EIII promoter expression was studied in coinfection or single-infection experiments. In both cases, transcription of the adenovirus early promoters was not stimulated by small-t. These and other results indicate that transactivation of polymerase II promoters by small-t occurs only when the target gene is in a transiently transfected state. Thus, small-t-mediated transactivation of polymerase II promoters is dependent on the type of assay system used and may be mechanistically different from that of the widely studied EIA.

Full text

PDF
6553

Images in this article

6555Image
on p.6555
6555Image
on p.6555
6556Image
on p.6556
6557Image
on p.6557
6559Image
on p.6559
6559Image
on p.6559
6559Image
on p.6559

Selected References

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

  1. Berger J., Hauber J., Hauber R., Geiger R., Cullen B. R. Secreted placental alkaline phosphatase: a powerful new quantitative indicator of gene expression in eukaryotic cells. Gene. 1988 Jun 15;66(1):1–10. doi: 10.1016/0378-1119(88)90219-3. [DOI] [PubMed] [Google Scholar]
  2. Berkner K. L., Schaffhausen B. S., Roberts T. M., Sharp P. A. Abundant expression of polyomavirus middle T antigen and dihydrofolate reductase in an adenovirus recombinant. J Virol. 1987 Apr;61(4):1213–1220. doi: 10.1128/jvi.61.4.1213-1220.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bhat B. M., Wold W. S. Genetic analysis of mRNA synthesis in adenovirus region E3 at different stages of productive infection by RNA-processing mutants. J Virol. 1986 Oct;60(1):54–63. doi: 10.1128/jvi.60.1.54-63.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bikel I., Montano X., Agha M. E., Brown M., McCormack M., Boltax J., Livingston D. M. SV40 small t antigen enhances the transformation activity of limiting concentrations of SV40 large T antigen. Cell. 1987 Jan 30;48(2):321–330. doi: 10.1016/0092-8674(87)90435-1. [DOI] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Cann A. J., Rosenblatt J. D., Wachsman W., Shah N. P., Chen I. S. Identification of the gene responsible for human T-cell leukaemia virus transcriptional regulation. Nature. 1985 Dec 12;318(6046):571–574. doi: 10.1038/318571a0. [DOI] [PubMed] [Google Scholar]
  8. Carey M. F., Singh K., Botchan M., Cozzarelli N. R. Induction of specific transcription by RNA polymerase III in transformed cells. Mol Cell Biol. 1986 Sep;6(9):3068–3076. doi: 10.1128/mcb.6.9.3068. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cullen B. R., Greene W. C. Regulatory pathways governing HIV-1 replication. Cell. 1989 Aug 11;58(3):423–426. doi: 10.1016/0092-8674(89)90420-0. [DOI] [PubMed] [Google Scholar]
  10. Davidson D., Hassell J. A. Overproduction of polyomavirus middle T antigen in mammalian cells through the use of an adenovirus vector. J Virol. 1987 Apr;61(4):1226–1239. doi: 10.1128/jvi.61.4.1226-1239.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Evans M. J., Scarpulla R. C. Both upstream and intron sequence elements are required for elevated expression of the rat somatic cytochrome c gene in COS-1 cells. Mol Cell Biol. 1988 Jan;8(1):35–41. doi: 10.1128/mcb.8.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Flint J., Shenk T. Adenovirus E1A protein paradigm viral transactivator. Annu Rev Genet. 1989;23:141–161. doi: 10.1146/annurev.ge.23.120189.001041. [DOI] [PubMed] [Google Scholar]
  13. Gaynor R. B., Tsukamoto A., Montell C., Berk A. J. Enhanced expression of adenovirus transforming proteins. J Virol. 1982 Oct;44(1):276–285. doi: 10.1128/jvi.44.1.276-285.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gingeras T. R., Sciaky D., Gelinas R. E., Bing-Dong J., Yen C. E., Kelly M. M., Bullock P. A., Parsons B. L., O'Neill K. E., Roberts R. J. Nucleotide sequences from the adenovirus-2 genome. J Biol Chem. 1982 Nov 25;257(22):13475–13491. [PubMed] [Google Scholar]
  15. Gluzman Y. SV40-transformed simian cells support the replication of early SV40 mutants. Cell. 1981 Jan;23(1):175–182. doi: 10.1016/0092-8674(81)90282-8. [DOI] [PubMed] [Google Scholar]
  16. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Graessmann A., Graessmann M., Tjian R., Topp W. C. Simian virus 40 small-t protein is required for loss of actin cable networks in rat cells. J Virol. 1980 Mar;33(3):1182–1191. doi: 10.1128/jvi.33.3.1182-1191.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hardy S., Engel D. A., Shenk T. An adenovirus early region 4 gene product is required for induction of the infection-specific form of cellular E2F activity. Genes Dev. 1989 Jul;3(7):1062–1074. doi: 10.1101/gad.3.7.1062. [DOI] [PubMed] [Google Scholar]
  19. Herrmann C. H., Mathews M. B. The adenovirus E1B 19-kilodalton protein stimulates gene expression by increasing DNA levels. Mol Cell Biol. 1989 Dec;9(12):5412–5423. doi: 10.1128/mcb.9.12.5412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hiebert S. W., Lipp M., Nevins J. R. E1A-dependent trans-activation of the human MYC promoter is mediated by the E2F factor. Proc Natl Acad Sci U S A. 1989 May;86(10):3594–3598. doi: 10.1073/pnas.86.10.3594. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hiscott J., Wong A., Alper D., Xanthoudakis S. trans activation of type 1 interferon promoters by simian virus 40 T antigen. Mol Cell Biol. 1988 Aug;8(8):3397–3405. doi: 10.1128/mcb.8.8.3397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Huang M. M., Hearing P. The adenovirus early region 4 open reading frame 6/7 protein regulates the DNA binding activity of the cellular transcription factor, E2F, through a direct complex. Genes Dev. 1989 Nov;3(11):1699–1710. doi: 10.1101/gad.3.11.1699. [DOI] [PubMed] [Google Scholar]
  23. Inoue J., Seiki M., Taniguchi T., Tsuru S., Yoshida M. Induction of interleukin 2 receptor gene expression by p40x encoded by human T-cell leukemia virus type 1. EMBO J. 1986 Nov;5(11):2883–2888. doi: 10.1002/j.1460-2075.1986.tb04583.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Jones N., Shenk T. An adenovirus type 5 early gene function regulates expression of other early viral genes. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3665–3669. doi: 10.1073/pnas.76.8.3665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kekulé A. S., Lauer U., Meyer M., Caselmann W. H., Hofschneider P. H., Koshy R. The preS2/S region of integrated hepatitis B virus DNA encodes a transcriptional transactivator. Nature. 1990 Feb 1;343(6257):457–461. doi: 10.1038/343457a0. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. 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]
  28. Manohar C. F., Kratochvil J., Thimmapaya B. The adenovirus EII early promoter has multiple EIA-sensitive elements, two of which function cooperatively in basal and virus-induced transcription. J Virol. 1990 Jun;64(6):2457–2466. doi: 10.1128/jvi.64.6.2457-2466.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Murthy S. C., Bhat G. P., Thimmappaya B. Adenovirus EIIA early promoter: transcriptional control elements and induction by the viral pre-early EIA gene, which appears to be sequence independent. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2230–2234. doi: 10.1073/pnas.82.8.2230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nevins J. R. Induction of the synthesis of a 70,000 dalton mammalian heat shock protein by the adenovirus E1A gene product. Cell. 1982 Jul;29(3):913–919. doi: 10.1016/0092-8674(82)90453-6. [DOI] [PubMed] [Google Scholar]
  31. Nevins J. R. Regulation of early adenovirus gene expression. Microbiol Rev. 1987 Dec;51(4):419–430. doi: 10.1128/mr.51.4.419-430.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pallas D. C., Shahrik L. K., Martin B. L., Jaspers S., Miller T. B., Brautigan D. L., Roberts T. M. Polyoma small and middle T antigens and SV40 small t antigen form stable complexes with protein phosphatase 2A. Cell. 1990 Jan 12;60(1):167–176. doi: 10.1016/0092-8674(90)90726-u. [DOI] [PubMed] [Google Scholar]
  33. Phillips B., Rundell K. Failure of simian virus 40 small t antigen to disorganize actin cables in nonpermissive cell lines. J Virol. 1988 Mar;62(3):768–775. doi: 10.1128/jvi.62.3.768-775.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rundell K. Complete interaction of cellular 56,000- and 32,000-Mr proteins with simian virus 40 small-t antigen in productively infected cells. J Virol. 1987 Apr;61(4):1240–1243. doi: 10.1128/jvi.61.4.1240-1243.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schaffhausen B. S., Bockus B. J., Berkner K. L., Kaplan D., Roberts T. M. Characterization of middle T antigen expressed by using an adenovirus expression system. J Virol. 1987 Apr;61(4):1221–1225. doi: 10.1128/jvi.61.4.1221-1225.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Segawa K., Yamaguchi N. Induction of c-Ha-ras transcription in rat cells by simian virus 40 large T antigen. Mol Cell Biol. 1987 Jan;7(1):556–559. doi: 10.1128/mcb.7.1.556. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sekulovich R. E., Leary K., Sandri-Goldin R. M. The herpes simplex virus type 1 alpha protein ICP27 can act as a trans-repressor or a trans-activator in combination with ICP4 and ICP0. J Virol. 1988 Dec;62(12):4510–4522. doi: 10.1128/jvi.62.12.4510-4522.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Shenk T. E., Carbon J., Berg P. Construction and analysis of viable deletion mutants of simian virus 40. J Virol. 1976 May;18(2):664–671. doi: 10.1128/jvi.18.2.664-671.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Stein R., Ziff E. B. HeLa cell beta-tubulin gene transcription is stimulated by adenovirus 5 in parallel with viral early genes by an E1a-dependent mechanism. Mol Cell Biol. 1984 Dec;4(12):2792–2801. doi: 10.1128/mcb.4.12.2792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Stow N. D. Cloning of a DNA fragment from the left-hand terminus of the adenovirus type 2 genome and its use in site-directed mutagenesis. J Virol. 1981 Jan;37(1):171–180. doi: 10.1128/jvi.37.1.171-180.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Subramanian S., Bhat R. A., Rundell M. K., Thimmappaya B. Suppression of the translation defect phenotype specific for a virus-associated RNA-deficient adenovirus mutant in monkey cells by simian virus 40. J Virol. 1986 Nov;60(2):363–368. doi: 10.1128/jvi.60.2.363-368.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Thimmappaya B., Jones N., Shenk T. A mutation which alters initiation of transcription by RNA polymerase III on the Ad5 chromosome. Cell. 1979 Dec;18(4):947–954. doi: 10.1016/0092-8674(79)90207-1. [DOI] [PubMed] [Google Scholar]
  43. Thummel C., Tjian R., Hu S. L., Grodzicker T. Translational control of SV40 T antigen expressed from the adenovirus late promoter. Cell. 1983 Jun;33(2):455–464. doi: 10.1016/0092-8674(83)90427-0. [DOI] [PubMed] [Google Scholar]
  44. Triezenberg S. J., Kingsbury R. C., McKnight S. L. Functional dissection of VP16, the trans-activator of herpes simplex virus immediate early gene expression. Genes Dev. 1988 Jun;2(6):718–729. doi: 10.1101/gad.2.6.718. [DOI] [PubMed] [Google Scholar]
  45. Twu J. S., Schloemer R. H. Transcriptional trans-activating function of hepatitis B virus. J Virol. 1987 Nov;61(11):3448–3453. doi: 10.1128/jvi.61.11.3448-3453.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Walter G., Ruediger R., Slaughter C., Mumby M. Association of protein phosphatase 2A with polyoma virus medium tumor antigen. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2521–2525. doi: 10.1073/pnas.87.7.2521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. White E., Cipriani R. Role of adenovirus E1B proteins in transformation: altered organization of intermediate filaments in transformed cells that express the 19-kilodalton protein. Mol Cell Biol. 1990 Jan;10(1):120–130. doi: 10.1128/mcb.10.1.120. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES