Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1986 Jun;6(6):1875–1885. doi: 10.1128/mcb.6.6.1875

Sequences involved in initiation of simian virus 40 late transcription in the absence of T antigen.

F Omilli, M Ernoult-Lange, J Borde, E May
PMCID: PMC367725  PMID: 3023909

Abstract

We analyzed the sequences involved in vivo in the initiation of simian virus 40 (SV40) late transcription occurring in the absence of both SV40 origin sequences and T antigen. The constituent elements of the SV40 late promoters have already been the subject of extensive studies. In vitro studies have resulted in the description of two putative domains of the late promoters. The first domain consists of an 11-base-pair (bp) sequence, 5'-GGTACCTAACC-3', located 25 nucleotides (nt) upstream of the SV40 major late initiation site (MLIS) (J. Brady, M. Radonovich, M. Vodkin, V. Natarajan, M. Thoren, G. Das, J. Janik, and N. P. Salzman, Cell 31:624-633, 1982). The second domain is located within the G-C-rich region (J. Brady, M. Radonovich, M. Thoren, G. Das, and N. P. Salzman, Mol. Cell. Biol. 4:133-141; U. Hansen and P. A. Sharp, EMBO J. 2:2293-2303). Our previous in vivo studies permitted us to define a domain of the late promoter which extends from nt 332 to nt 113 and includes the 72-bp enhancer sequences. Here, by using transfection of the appropriate chimeric plasmids into HeLa cells in conjunction with quantitative S1 nuclease analysis, we analyzed in more detail the sequences required for the control of SV40 late-gene expression occurring before the onset of viral DNA replication. We showed that the major late promoter element is in fact the 72-bp repeat enhancer element. This element was able to drive efficient late transcription in the absence of T antigen. Under our experimental conditions, removal of the G-C-rich region (21-bp repeats) entailed a significant increase in the level of late-gene expression. Moreover, translocation of this element closer to the MLIS (53 nt upstream of the MLIS) enhanced the level of transcripts initiated at natural late initiation sites. Our results suggest that the G-C-rich regions have to be positioned between the enhancer element and the initiation sites to stimulate transcription from downstream sites. Thus, the relative arrangement of the various promoter elements is a critical factor contributing to the situation in which the early promoter is stronger than late promoters before viral DNA replication.

Full text

PDF
1875

Images in this article

1880Image
on p.1880
1881Image
on p.1881
1882Image
on p.1882

Selected References

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

  1. Alwine J. C. Transient gene expression control: effects of transfected DNA stability and trans-activation by viral early proteins. Mol Cell Biol. 1985 May;5(5):1034–1042. doi: 10.1128/mcb.5.5.1034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Banerji J., Rusconi S., Schaffner W. Expression of a beta-globin gene is enhanced by remote SV40 DNA sequences. Cell. 1981 Dec;27(2 Pt 1):299–308. doi: 10.1016/0092-8674(81)90413-x. [DOI] [PubMed] [Google Scholar]
  3. Benoist C., Chambon P. In vivo sequence requirements of the SV40 early promotor region. Nature. 1981 Mar 26;290(5804):304–310. doi: 10.1038/290304a0. [DOI] [PubMed] [Google Scholar]
  4. Berk A. J., Sharp P. A. Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274–1278. doi: 10.1073/pnas.75.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brady J., Khoury G. trans Activation of the simian virus 40 late transcription unit by T-antigen. Mol Cell Biol. 1985 Jun;5(6):1391–1399. doi: 10.1128/mcb.5.6.1391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brady J., Radonovich M., Thoren M., Das G., Salzman N. P. Simian virus 40 major late promoter: an upstream DNA sequence required for efficient in vitro transcription. Mol Cell Biol. 1984 Jan;4(1):133–141. doi: 10.1128/mcb.4.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brady J., Radonovich M., Vodkin M., Natarajan V., Thoren M., Das G., Janik J., Salzman N. P. Site-specific base substitution and deletion mutations that enhance or suppress transcription of the SV40 major late RNA. Cell. 1982 Dec;31(3 Pt 2):625–633. doi: 10.1016/0092-8674(82)90318-x. [DOI] [PubMed] [Google Scholar]
  8. Buchman A. R., Fromm M., Berg P. Complex regulation of simian virus 40 early-region transcription from different overlapping promoters. Mol Cell Biol. 1984 Sep;4(9):1900–1914. doi: 10.1128/mcb.4.9.1900. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Contreras R., Fiers W. Initiation of transcription by RNA polymerase II in permeable, SV40-infected or noninfected, CVI cells; evidence for multiple promoters of SV40 late transcription. Nucleic Acids Res. 1981 Jan 24;9(2):215–236. doi: 10.1093/nar/9.2.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dynan W. S., Tjian R. Isolation of transcription factors that discriminate between different promoters recognized by RNA polymerase II. Cell. 1983 Mar;32(3):669–680. doi: 10.1016/0092-8674(83)90053-3. [DOI] [PubMed] [Google Scholar]
  11. Dynan W. S., Tjian R. The promoter-specific transcription factor Sp1 binds to upstream sequences in the SV40 early promoter. Cell. 1983 Nov;35(1):79–87. doi: 10.1016/0092-8674(83)90210-6. [DOI] [PubMed] [Google Scholar]
  12. Ernoult-Lange M., May E. Evidence of transcription from the late region of the integrated simian virus 40 genome in transformed cells: location of the 5' ends of late transcripts in cells abortively infected and in cells transformed by simian virus 40. J Virol. 1983 Jun;46(3):756–767. doi: 10.1128/jvi.46.3.756-767.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ernoult-Lange M., May P., Moreau P., May E. Simian virus 40 late promoter region able to initiate simian virus 40 early gene transcription in the absence of the simian virus 40 origin sequence. J Virol. 1984 Apr;50(1):163–173. doi: 10.1128/jvi.50.1.163-173.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Everett R. D., Baty D., Chambon P. The repeated GC-rich motifs upstream from the TATA box are important elements of the SV40 early promoter. Nucleic Acids Res. 1983 Apr 25;11(8):2447–2464. doi: 10.1093/nar/11.8.2447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fromm M., Berg P. Deletion mapping of DNA regions required for SV40 early region promoter function in vivo. J Mol Appl Genet. 1982;1(5):457–481. [PubMed] [Google Scholar]
  16. Ghosh P. K., Lebowitz P. Simian virus 40 early mRNA's contain multiple 5' termini upstream and downstream from a Hogness-Goldberg sequence; a shift in 5' termini during the lytic cycle is mediated by large T antigen. J Virol. 1981 Oct;40(1):224–240. doi: 10.1128/jvi.40.1.224-240.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ghosh P. K., Reddy V. B., Swinscoe J., Lebowitz P., Weissman S. M. Heterogeneity and 5'-terminal structures of the late RNAs of simian virus 40. J Mol Biol. 1978 Dec 25;126(4):813–846. doi: 10.1016/0022-2836(78)90022-0. [DOI] [PubMed] [Google Scholar]
  18. Gruss P., Dhar R., Khoury G. Simian virus 40 tandem repeated sequences as an element of the early promoter. Proc Natl Acad Sci U S A. 1981 Feb;78(2):943–947. doi: 10.1073/pnas.78.2.943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Haegeman G., Fiers W. Localization of the 5' terminus of late SV40 mRNA. Nucleic Acids Res. 1978 Jul;5(7):2359–2371. doi: 10.1093/nar/5.7.2359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hansen U., Sharp P. A. Sequences controlling in vitro transcription of SV40 promoters. EMBO J. 1983;2(12):2293–2303. doi: 10.1002/j.1460-2075.1983.tb01737.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lange M., May E., May P. Ability of nonpermissive mouse cells to express a simian virus 40 late function(s). J Virol. 1981 Jun;38(3):940–951. doi: 10.1128/jvi.38.3.940-951.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  23. Mishoe H., Brady J. N., Radonovich M., Salzman N. P. Simian virus 40 guanine-cytosine-rich sequences function as independent transcriptional control elements in vitro. Mol Cell Biol. 1984 Dec;4(12):2911–2920. doi: 10.1128/mcb.4.12.2911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Moreau P., Hen R., Wasylyk B., Everett R., Gaub M. P., Chambon P. The SV40 72 base repair repeat has a striking effect on gene expression both in SV40 and other chimeric recombinants. Nucleic Acids Res. 1981 Nov 25;9(22):6047–6068. doi: 10.1093/nar/9.22.6047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nandi A., Das G., Salzman N. P. Characterization of a surrogate TATA box promoter that regulates in vitro transcription of the simian virus 40 major late gene. Mol Cell Biol. 1985 Mar;5(3):591–594. doi: 10.1128/mcb.5.3.591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Reddy V. B., Ghosh P. K., Lebowitz P., Weissman S. M. Gaps and duplicated sequences in the leaders of SV40 16S RNA. Nucleic Acids Res. 1978 Nov;5(11):4195–4213. doi: 10.1093/nar/5.11.4195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rio D. C., Tjian R. Multiple control elements involved in the initiation of SV40 late transcription. J Mol Appl Genet. 1984;2(5):423–435. [PubMed] [Google Scholar]
  28. Sassone-Corsi P., Dougherty J. P., Wasylyk B., Chambon P. Stimulation of in vitro transcription from heterologous promoters by the simian virus 40 enhancer. Proc Natl Acad Sci U S A. 1984 Jan;81(2):308–312. doi: 10.1073/pnas.81.2.308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Vigneron M., Barrera-Saldana H. A., Baty D., Everett R. E., Chambon P. Effect of the 21-bp repeat upstream element on in vitro transcription from the early and late SV40 promoters. EMBO J. 1984 Oct;3(10):2373–2382. doi: 10.1002/j.1460-2075.1984.tb02142.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wasylyk B., Wasylyk C., Augereau P., Chambon P. The SV40 72 bp repeat preferentially potentiates transcription starting from proximal natural or substitute promoter elements. Cell. 1983 Feb;32(2):503–514. doi: 10.1016/0092-8674(83)90470-1. [DOI] [PubMed] [Google Scholar]
  31. Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weiher H., König M., Gruss P. Multiple point mutations affecting the simian virus 40 enhancer. Science. 1983 Feb 11;219(4585):626–631. doi: 10.1126/science.6297005. [DOI] [PubMed] [Google Scholar]
  33. de Villiers J., Olson L., Banerji J., Schaffner W. Analysis of the transcriptional enhancer effect. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 2):911–919. doi: 10.1101/sqb.1983.047.01.105. [DOI] [PubMed] [Google Scholar]

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

RESOURCES