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. 1986 Jan;57(1):91–100. doi: 10.1128/jvi.57.1.91-100.1986

Expression of the late gene of simian virus 40 under the control of the simian virus 40 early-region promoter in monkey and mouse cells.

V N Rao, P K Ghosh, S M Weissman
PMCID: PMC252702  PMID: 3001368

Abstract

We constructed a recombinant plasmid (pVNR4) with the simian virus 40 (SV40) early promoter positioned 30 nucleotides upstream from the major SV40 late transcription initiation site at residue 325. After transfection of the recombinant plasmid DNA into COS and mouse L cells, the transcripts of the SV40 late region were analyzed by S1 nuclease and primer extension analysis. The following are the principal findings. (i) The 16S and 19S late RNAs used the characteristic wild-type splice; no detectable levels of 19S unspliced RNA were observed. (ii) The majority of the late RNAs were heterogeneous and initiated in the early region (upstream and downstream from the Hogness-Goldberg sequence), and a minor population initiated at residue 325, the principal 5' terminus of the wild-type late RNA. (iii) During SV40 lytic infection there was a shift in initiation sites used to transcribe the early region from sites that are downstream to sites which are upstream (up RNA) of the origin of DNA replication. We observed that unlike lytic infection, T antigen and viral DNA replication were not needed for the appearance of up RNA in mouse L cells. (iv) In mouse L cells late RNAs were made, and the residue 325 5' end was utilized in the absence of T antigen or DNA replication. (v) In COS cells we found down RNA and up RNA transcribed from the extrachromosomally replicating plasmid but only down RNA produced by the integrated SV40 genome.

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Selected References

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

  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barkan A., Mertz J. E. DNA sequence analysis of simian virus 40 mutants with deletions mapping in the leader region of the late viral mRNA's: mutants with deletions similar in size and position exhibit varied phenotypes. J Virol. 1981 Feb;37(2):730–737. doi: 10.1128/jvi.37.2.730-737.1981. [DOI] [PMC free article] [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. Bergsma D. J., Olive D. M., Hartzell S. W., Subramanian K. N. Territorial limits and functional anatomy of the simian virus 40 replication origin. Proc Natl Acad Sci U S A. 1982 Jan;79(2):381–385. doi: 10.1073/pnas.79.2.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  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. Enquist L. W., Vande Woude G. F., Wagner M., Smiley J. R., Summers W. C. Construction and characterization of a recombinant plasmid encoding the gene for the thymidine kinase of Herpes simplex type 1 virus. Gene. 1979 Nov;7(3-4):335–342. doi: 10.1016/0378-1119(79)90052-0. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. 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]
  13. Ganem D., Nussbaum A. L., Davoli D., Fareed G. C. Propagation of a segment of bacteriophage lamda-DNA in monkey cells after covalent linkage to a defective simian virus 40 genome. Cell. 1976 Mar;7(3):349–359. doi: 10.1016/0092-8674(76)90164-1. [DOI] [PubMed] [Google Scholar]
  14. Ghosh P. K., Lebowitz P., Frisque R. J., Gluzman Y. Identification of a promoter component involved in positioning the 5' termini of simian virus 40 early mRNAs. Proc Natl Acad Sci U S A. 1981 Jan;78(1):100–104. doi: 10.1073/pnas.78.1.100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Ghosh P. K., Piatak M., Mertz J. E., Weissman S. M., Lebowitz P. Altered utilization of splice sites and 5' termini in late RNAs produced by leader region mutants of simian virus 40. J Virol. 1982 Nov;44(2):610–624. doi: 10.1128/jvi.44.2.610-624.1982. [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. Ghosh P. K., Roy P., Barkan A., Mertz J. E., Weissman S. M., Lebowitz P. Unspliced functional late 19S mRNAs containing intervening sequences are produced by a late leader mutant of simian virus 40. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1386–1390. doi: 10.1073/pnas.78.3.1386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Goff S. P., Berg P. Construction of hybrid viruses containing SV40 and lambda phage DNA segments and their propagation in cultured monkey cells. Cell. 1976 Dec;9(4 Pt 2):695–705. doi: 10.1016/0092-8674(76)90133-1. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Gruss P., Khoury G. Expression of simian virus 40-rat preproinsulin recombinants in monkey kidney cells: use of preproinsulin RNA processing signals. Proc Natl Acad Sci U S A. 1981 Jan;78(1):133–137. doi: 10.1073/pnas.78.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Haegeman G., Fiers W. Characterization of the 5'-terminal capped structures of late simian virus 40-specific mRNA. J Virol. 1978 Mar;25(3):824–830. doi: 10.1128/jvi.25.3.824-830.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Haegeman G., van Heuverswyn H., Gheysen D., Fiers W. Heterogeneity of the 5' terminus of late mRNA induced by a viable simian virus 40 deletion mutant. J Virol. 1979 Aug;31(2):484–493. doi: 10.1128/jvi.31.2.484-493.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Hamer D. H., Kaehler M., Leder P. A mouse globin gene promoter is functional in SV40. Cell. 1980 Oct;21(3):697–708. doi: 10.1016/0092-8674(80)90433-x. [DOI] [PubMed] [Google Scholar]
  26. Hamer D. H., Leder P. Expression of the chromosomal mouse Beta maj-globin gene cloned in SV40. Nature. 1979 Sep 6;281(5726):35–40. doi: 10.1038/281035a0. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Hay R. T., DePamphilis M. L. Initiation of SV40 DNA replication in vivo: location and structure of 5' ends of DNA synthesized in the ori region. Cell. 1982 Apr;28(4):767–779. doi: 10.1016/0092-8674(82)90056-3. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Horowitz M., Cepko C. L., Sharp P. A. Expression of chimeric genes in the early region of SV40. J Mol Appl Genet. 1983;2(2):147–159. [PubMed] [Google Scholar]
  31. 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]
  32. Khoury G., May E. Regulation of early and late simian virus 40 transcription: overproduction of early viral RNA in the absence of a functional T-antigen. J Virol. 1977 Jul;23(1):167–176. doi: 10.1128/jvi.23.1.167-176.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Lai C. J., Dhar R., Khoury G. Mapping the spliced and unspliced late lytic SV40 RNAs. Cell. 1978 Aug;14(4):971–982. doi: 10.1016/0092-8674(78)90351-3. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Lazarowitz S. G. Simian virus 40 mutant with transposed T-antigen and VP1 genes. J Virol. 1982 Mar;41(3):1025–1037. doi: 10.1128/jvi.41.3.1025-1037.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Lebowitz P., Weissman S. M. Organization and transcription of the simian virus 40 genome. Curr Top Microbiol Immunol. 1979;87:43–172. doi: 10.1007/978-3-642-67344-3_3. [DOI] [PubMed] [Google Scholar]
  37. Lusky M., Botchan M. Inhibition of SV40 replication in simian cells by specific pBR322 DNA sequences. Nature. 1981 Sep 3;293(5827):79–81. doi: 10.1038/293079a0. [DOI] [PubMed] [Google Scholar]
  38. Martin R. G. The transformation of cell growth and transmogrification of DNA synthesis by simian virus 40. Adv Cancer Res. 1981;34:1–68. doi: 10.1016/s0065-230x(08)60238-9. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Mertz J. E., Berg P. Viable deletion mutants of simian virus 40: selective isolation by means of a restriction endonuclease from Hemophilus parainfluenzae. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4879–4883. doi: 10.1073/pnas.71.12.4879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. 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]
  42. Mulligan R. C., Howard B. H., Berg P. Synthesis of rabbit beta-globin in cultured monkey kidney cells following infection with a SV40 beta-globin recombinant genome. Nature. 1979 Jan 11;277(5692):108–114. doi: 10.1038/277108a0. [DOI] [PubMed] [Google Scholar]
  43. 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]
  44. Piatak M., Ghosh P. K., Norkin L. C., Weissman S. M. Sequences locating the 5' ends of the major simian virus 40 late mRNA forms. J Virol. 1983 Nov;48(2):503–520. doi: 10.1128/jvi.48.2.503-520.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Piatak M., Subramanian K. N., Roy P., Weissman S. M. Late messenger RNA production by viable simian virus 40 mutants with deletions in the leader region. J Mol Biol. 1981 Dec 15;153(3):589–618. doi: 10.1016/0022-2836(81)90409-5. [DOI] [PubMed] [Google Scholar]
  46. Reddy V. B., Ghosh P. K., Lebowitz P., Piatak M., Weissman S. M. Simian virus 40 early mRNA's. I. Genomic localization of 3' and 5' termini and two major splices in mRNA from transformed and lytically infected cells. J Virol. 1979 Apr;30(1):279–296. doi: 10.1128/jvi.30.1.279-296.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. 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]
  48. Reddy V. B., Tevethia S. S., Tevethia M. J., Weissman S. M. Nonselective expression of simian virus 40 large tumor antigen fragments in mouse cells. Proc Natl Acad Sci U S A. 1982 Mar;79(6):2064–2067. doi: 10.1073/pnas.79.6.2064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Reddy V. B., Thimmappaya B., Dhar R., Subramanian K. N., Zain B. S., Pan J., Ghosh P. K., Celma M. L., Weissman S. M. The genome of simian virus 40. Science. 1978 May 5;200(4341):494–502. doi: 10.1126/science.205947. [DOI] [PubMed] [Google Scholar]
  50. 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]
  51. 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]
  52. Subramanian K. N. Segments of simian virus 40 DNA spanning most of the leader sequence of the major late viral messenger RNA are dispensable. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2556–2560. doi: 10.1073/pnas.76.6.2556. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Tegtmeyer P., Schwartz M., Collins J. K., Rundell K. Regulation of tumor antigen synthesis by simain virus 40 gene A. J Virol. 1975 Jul;16(1):168–178. doi: 10.1128/jvi.16.1.168-178.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Twigg A. J., Sherratt D. Trans-complementable copy-number mutants of plasmid ColE1. Nature. 1980 Jan 10;283(5743):216–218. doi: 10.1038/283216a0. [DOI] [PubMed] [Google Scholar]
  55. Villarreal L. P., White R. T., Berg P. Mutational alterations within the simian virus 40 leader segment generate altered 16S and 19S mRNA's. J Virol. 1979 Jan;29(1):209–219. doi: 10.1128/jvi.29.1.209-219.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Wasylyk B., Wasylyk C., Matthes H., Wintzerith M., Chambon P. Transcription from the SV40 early-early and late-early overlapping promoters in the absence of DNA replication. EMBO J. 1983;2(9):1605–1611. doi: 10.1002/j.1460-2075.1983.tb01631.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Weinberg R. A., Ben-Ishai Z., Newbold J. E. Simian virus 40 transcription in productively infected and transformed cells. J Virol. 1974 Jun;13(6):1263–1273. doi: 10.1128/jvi.13.6.1263-1273.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Wigler M., Pellicer A., Silverstein S., Axel R., Urlaub G., Chasin L. DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1373–1376. doi: 10.1073/pnas.76.3.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]

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