Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 May;79(9):2743–2747. doi: 10.1073/pnas.79.9.2743

Site of premature termination of late transcription of simian virus 40 DNA: enhancement by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole.

H Skolnik-David, N Hay, Y Aloni
PMCID: PMC346282  PMID: 6283541

Abstract

Sedimentation analysis of pulse-labeled RNA synthesized in nuclei isolated from simian virus 40-infected cells revealed an abundance of short cellular and viral RNAs. The relative amount of the short chains is increased in nuclei isolated from cells treated with 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB). The short viral RNAs were purified by hybridization to and elution from simian virus 40 DNA on filters, and their sizes were determined by gel electrophoresis. A major band of 93- to 95-nucleotide-long RNA was observed along with additional minor bands. Identical bands were revealed when the viral RNA was purified from nuclei of cells pretreated with DRB. The major band was identified as an aborted transcript of a RNA that initiated at the major initiation site (nucleotide 243). We have found that the DNA region where the RNA stops is A+T rich and is immediately preceded by a G+C-rich region that exhibits dyad symmetry, resembling the termination signal in prokaryotes. These observations show that RNA polymerase II responds to the same termination signal as the prokaryotic enzyme and suggest that a mechanism of attenuation regulates simian virus 40 late transcription.

Full text

PDF
2743

Images in this article

Selected References

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

  1. Adhya S., Gottesman M. Control of transcription termination. Annu Rev Biochem. 1978;47:967–996. doi: 10.1146/annurev.bi.47.070178.004535. [DOI] [PubMed] [Google Scholar]
  2. Aloni Y., Bratosin S., Dhar R., Laub O., Horowitz M., Khoury G. Splicing of SV40 mRNAs: a novel mechanism for the regulation of gene expression in animal cells. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):559–570. doi: 10.1101/sqb.1978.042.01.058. [DOI] [PubMed] [Google Scholar]
  3. Aloni Y. Splicing of viral mRNAs. Prog Nucleic Acid Res Mol Biol. 1981;25:1–31. doi: 10.1016/s0079-6603(08)60481-x. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Benz E. W., Jr, Wydro R. M., Nadal-Ginard B., Dina D. Moloney murine sarcoma proviral DNA is a transcriptional unit. Nature. 1980 Dec 25;288(5792):665–669. doi: 10.1038/288665a0. [DOI] [PubMed] [Google Scholar]
  6. Bertrand K., Korn L., Lee F., Platt T., Squires C. L., Squires C., Yanofsky C. New features of the regulation of the tryptophan operon. Science. 1975 Jul 4;189(4196):22–26. doi: 10.1126/science.1094538. [DOI] [PubMed] [Google Scholar]
  7. Canaani D., Kahana C., Mukamel A., Groner Y. Sequence heterogeneity at the 5' termini of late simian virus 40 19S and 16S mRNAs. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3078–3082. doi: 10.1073/pnas.76.7.3078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Crawford I. P., Stauffer G. V. Regulation of tryptophan biosynthesis. Annu Rev Biochem. 1980;49:163–195. doi: 10.1146/annurev.bi.49.070180.001115. [DOI] [PubMed] [Google Scholar]
  10. Evans R., Weber J., Ziff E., Darnell J. E. Premature termination during adenovirus transcription. Nature. 1979 Mar 22;278(5702):367–370. doi: 10.1038/278367a0. [DOI] [PubMed] [Google Scholar]
  11. Fiers W., Contreras R., Haegemann G., Rogiers R., Van de Voorde A., Van Heuverswyn H., Van Herreweghe J., Volckaert G., Ysebaert M. Complete nucleotide sequence of SV40 DNA. Nature. 1978 May 11;273(5658):113–120. doi: 10.1038/273113a0. [DOI] [PubMed] [Google Scholar]
  12. Fraser N. W., Sehgal P. B., Darnell J. E., Jr Multiple discrete sites for premature RNA chain termination late in adenovirus-2 infection: enhancement by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2571–2575. doi: 10.1073/pnas.76.6.2571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Frunzio R., Bruni C. B., Blasi F. In vivo and in vitro detection of the leader RNA of the histidine operon of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1981 May;78(5):2767–2771. doi: 10.1073/pnas.78.5.2767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Galluppi G. R., Richardson J. P. ATP-induced changes in the binding of RNA synthesis termination protein Rho to RNA. J Mol Biol. 1980 Apr 15;138(3):513–539. doi: 10.1016/s0022-2836(80)80016-7. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Gidoni D., Kahana C., Canaani D., Groner Y. Specific in vitro initiation of transcription of simian virus 40 early and late genes occurs at the various cap nucleotides including cytidine. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2174–2178. doi: 10.1073/pnas.78.4.2174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gottesman M. E., Adhya S., Das A. Transcription antitermination by bacteriophage lambda N gene product. J Mol Biol. 1980 Jun 15;140(1):57–75. doi: 10.1016/0022-2836(80)90356-3. [DOI] [PubMed] [Google Scholar]
  18. Haegeman G., Iserentant D., Gheysen D., Fiers W. Characterization of the major altered leader sequence of late mRNA induced by SV40 deletion mutant d1-1811. Nucleic Acids Res. 1979 Dec 11;7(7):1799–1814. doi: 10.1093/nar/7.7.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Horowitz M., Laub O., Bratosin S., Aloni Y. Splicing of SV40 late mRNA is a post-transcriptional process. Nature. 1978 Oct 12;275(5680):558–559. doi: 10.1038/275558a0. [DOI] [PubMed] [Google Scholar]
  20. Huang C. C., Hearst J. E. Pauses at positions of secondary structure during in vitro replication of single-stranded fd bacteriophage DNA by T4 DNA polymerase. Anal Biochem. 1980 Mar 15;103(1):127–139. doi: 10.1016/0003-2697(80)90246-8. [DOI] [PubMed] [Google Scholar]
  21. Jakobovits E. B., Bratosin S., Aloni Y. A nucleosome-free region in SV40 minichromosomes. Nature. 1980 May 22;285(5762):263–265. doi: 10.1038/285263a0. [DOI] [PubMed] [Google Scholar]
  22. Jay G., Nomura S., Anderson C. W., Khoury G. Identification of the SV40 agnogene product: a DNA binding protein. Nature. 1981 May 28;291(5813):346–349. doi: 10.1038/291346a0. [DOI] [PubMed] [Google Scholar]
  23. Korn L. J., Brown D. D. Nucleotide sequence of Xenopus borealis oocyte 5S DNA: comparison of sequences that flank several related eucaryotic genes. Cell. 1978 Dec;15(4):1145–1156. doi: 10.1016/0092-8674(78)90042-9. [DOI] [PubMed] [Google Scholar]
  24. Laub O., Aloni Y. Transcription of simian virus 40. V. Regulattion of simian virus 40 gene expression. J Virol. 1975 Nov;16(5):1171–1183. doi: 10.1128/jvi.16.5.1171-1183.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Laub O., Aloni Y. Transcription of simian virus 40. VI. SV 40 DNA-RNA polymerase complex isolated from productively infected cells transcribed in vitro. Virology. 1976 Dec;75(2):346–354. doi: 10.1016/0042-6822(76)90033-7. [DOI] [PubMed] [Google Scholar]
  26. Laub O., Bratosin S., Horowitz M., Aloni Y. The initiation of transcription of SV40 DNA at late time after infection. Virology. 1979 Jan 30;92(2):310–323. doi: 10.1016/0042-6822(79)90136-3. [DOI] [PubMed] [Google Scholar]
  27. Laub O., Jakobovits E. B., Aloni Y. 5,6-dichloro-1-beta-ribofuranosylbenzimidazole enhances premature termination of late transcription of simian virus 40 DNA. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3297–3301. doi: 10.1073/pnas.77.6.3297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Maniatis T., Efstratiadis A. Fractionation of low molecular weight DNA or RNA in polyacrylamide gels containing 98% formamide or 7 M urea. Methods Enzymol. 1980;65(1):299–305. doi: 10.1016/s0076-6879(80)65040-x. [DOI] [PubMed] [Google Scholar]
  30. Martin F. H., Tinoco I., Jr DNA-RNA hybrid duplexes containing oligo(dA:rU) sequences are exceptionally unstable and may facilitate termination of transcription. Nucleic Acids Res. 1980 May 24;8(10):2295–2299. doi: 10.1093/nar/8.10.2295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mills D. R., Dobkin C., Kramer F. R. Template-determined, variable rate of RNA chain elongation. Cell. 1978 Oct;15(2):541–550. doi: 10.1016/0092-8674(78)90022-3. [DOI] [PubMed] [Google Scholar]
  32. Penman S. RNA metabolism in the HeLa cell nucleus. J Mol Biol. 1966 May;17(1):117–130. doi: 10.1016/s0022-2836(66)80098-0. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Remington J. A. Attenuation as a general mechanism for the regulation of differential gene transcription in eukaryoties. FEBS Lett. 1979 Apr 15;100(2):225–229. doi: 10.1016/0014-5793(79)80339-7. [DOI] [PubMed] [Google Scholar]
  35. Robberson D., Aloni Y., Attardi G. Electron microscopic visualization of mitochondrial RNA-DNA hybrids. J Mol Biol. 1971 Jan 28;55(2):267–270. doi: 10.1016/0022-2836(71)90196-3. [DOI] [PubMed] [Google Scholar]
  36. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  37. Salditt-Georgieff M., Harpold M., Chen-Kiang S., Darnell J. E., Jr The addition of 5' cap structures occurs early in hnRNA synthesis and prematurely terminated molecules are capped. Cell. 1980 Jan;19(1):69–78. doi: 10.1016/0092-8674(80)90389-x. [DOI] [PubMed] [Google Scholar]
  38. Saragosti S., Moyne G., Yaniv M. Absence of nucleosomes in a fraction of SV40 chromatin between the origin of replication and the region coding for the late leader RNA. Cell. 1980 May;20(1):65–73. doi: 10.1016/0092-8674(80)90235-4. [DOI] [PubMed] [Google Scholar]
  39. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. Tamm I., Kikuchi T. Early termination of heterogeneous nuclear RNA transcripts in mammalian cells: accentuation by 5,6-dichloro 1-beta-D-ribofuranosylbenzimidazole. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5750–5754. doi: 10.1073/pnas.76.11.5750. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Testa D., Chanda P. K., Banerjee A. K. Unique mode of transcription in vitro by Vesicular stomatitis virus. Cell. 1980 Aug;21(1):267–275. doi: 10.1016/0092-8674(80)90134-8. [DOI] [PubMed] [Google Scholar]
  43. Veldman G. M., Klootwijk J., de Jonge P., Leer R. J., Planta R. J. The transcription termination site of the ribosomal RNA operon in yeast. Nucleic Acids Res. 1980 Nov 25;8(22):5179–5192. doi: 10.1093/nar/8.22.5179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wu A. M., Chapman A. B., Platt T., Guarente L. P., Beckwith J. Deletions of distal sequence after termination of transcription at the end of the tryptophan operon in E. coli. Cell. 1980 Apr;19(4):829–836. doi: 10.1016/0092-8674(80)90073-2. [DOI] [PubMed] [Google Scholar]
  45. Yanofsky C. Attenuation in the control of expression of bacterial operons. Nature. 1981 Feb 26;289(5800):751–758. doi: 10.1038/289751a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES