Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1979 Aug;31(2):437–446. doi: 10.1128/jvi.31.2.437-446.1979

Characterization of the 5'-terminal structure of simian virus 40 early mRNA's.

J A Thompson, M F Radonovich, N P Salzman
PMCID: PMC353467  PMID: 90173

Abstract

RPC-5 reverse-phase chromatography has been used to isolate fragments of simian virus 40 DNA generated by appropriate digestions with restriction endonucleases. Ten specific DNA fragments, mapping successively in a counterclock-wise direction from 0.67 to 0.515 on the simian virus 40 genome, were each hybridized to cytoplasmic mRNA obtained during the early phase of simian virus 40 infection. Primer extension methods with reverse transcriptase were used to characterize the 5' ends of two species of viral mRNA which were fractionated on sucrose gradients. Analysis of the complementary DNA products demonstrated the presence of two different spliced structures of simian virus 40 early mRNA's, both of which had the same 5'-end sequences (AUU), located at residues 18 to 20 on the viral genome. The mRNA for small-t contained a segment 588 bases in length (residues 18 to 605) spliced to residues 672. A 66-nucleotide segment rich in adenine-thymine was spliced out of this mRNA. The mRNA for large-T contained a segment 308 bases in length (residues 18 to 325) which is also spliced to residue 672. A 346-base segment was spliced from this mRNA. The results suggest that there are two levels for control of genetic expression. One would be the regulation of initiation of transcription at a common promoter; the other involves post-transcriptional splicing.

Full text

PDF
440

Images in this article

Selected References

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

  1. 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]
  2. Bina-Stein M., Thoren M., Salzman N., Thomspon J. A. Rapid sequence determination of late simian virus 40 16S mRNA leader by using inhibitors of reverse transcriptase. Proc Natl Acad Sci U S A. 1979 Feb;76(2):731–735. doi: 10.1073/pnas.76.2.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. CRAWFORD L. V., BLACK P. H. THE NUCLEIC ACID OF SIMIAN VIRUS 40. Virology. 1964 Nov;24:388–392. doi: 10.1016/0042-6822(64)90176-x. [DOI] [PubMed] [Google Scholar]
  4. Casey J., Davidson N. Rates of formation and thermal stabilities of RNA:DNA and DNA:DNA duplexes at high concentrations of formamide. Nucleic Acids Res. 1977;4(5):1539–1552. doi: 10.1093/nar/4.5.1539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Deeley R. G., Gordon J. I., Burns A. T., Mullinix K. P., Binastein M., Goldberg R. F. Primary activation of the vitellogenin gene in the rooster. J Biol Chem. 1977 Nov 25;252(22):8310–8319. [PubMed] [Google Scholar]
  6. Eshaghpour H., Crothers D. M. Preparative separation of the complementary strands of DNA restriction fragments by alkaline RPC-5 chromatography. Nucleic Acids Res. 1978 Jan;5(1):13–21. doi: 10.1093/nar/5.1.13. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Groner Y., Carmi P., Aloni Y. Capping structures of simian virus 40 19S and 16S mRNAs. Nucleic Acids Res. 1977 Nov;4(11):3959–3968. doi: 10.1093/nar/4.11.3959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Hardies S. C., Wells R. D. Preparative fractionation of DNA restriction fragments by reversed phase column chromatography. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3117–3121. doi: 10.1073/pnas.73.9.3117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Landy A., Foeller C., Reszelbach R., Dudock B. Preparative fractionation of DNA restriction fragments by high pressure column chromatography on RPC-5. Nucleic Acids Res. 1976 Oct;3(10):2575–2592. doi: 10.1093/nar/3.10.2575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lavi S., Shatkin A. J. Methylated simian virus 40-specific RNA from nuclei and cytoplasm of infected BSC-1 cells. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2012–2016. doi: 10.1073/pnas.72.6.2012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. May E., Kress M., May P. Characterization of two SV40 early mRNAs and evidence for a nuclear "prespliced" RNA species. Nucleic Acids Res. 1978 Sep;5(9):3083–3099. doi: 10.1093/nar/5.9.3083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McCarty K. S., Jr, Vollmer R. T., McCarty K. S. Improved computer program data for the resolution and fractionation of macromolecules by isokinetic sucrose density gradient sedimentation. Anal Biochem. 1974 Sep;61(1):165–183. doi: 10.1016/0003-2697(74)90343-1. [DOI] [PubMed] [Google Scholar]
  17. Paucha E., Smith A. E. The sequences between 0.59 and 0.54 map units on SV40 DNA code for the unique region of small t antigen. Cell. 1978 Nov;15(3):1011–1020. doi: 10.1016/0092-8674(78)90285-4. [DOI] [PubMed] [Google Scholar]
  18. Prives C., Gilboa E., Revel M., Winocour E. Cell-free translation of simian virus 40 early messenger RNA coding for viral T-antigen. Proc Natl Acad Sci U S A. 1977 Feb;74(2):457–461. doi: 10.1073/pnas.74.2.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sebring E. D., Kelly T. J., Jr, Thoren M. M., Salzman N. P. Structure of replicating simian virus 40 deoxyribonucleic acid molecules. J Virol. 1971 Oct;8(4):478–490. doi: 10.1128/jvi.8.4.478-490.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shani M., Birkenmeier E., May E., Salzman N. P. Properties of simian virus 40 transcriptional intermediates isolated from nuclei of permissive cells. J Virol. 1977 Jul;23(1):20–28. doi: 10.1128/jvi.23.1.20-28.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tilghman S. M., Tiemeier D. C., Seidman J. G., Peterlin B. M., Sullivan M., Maizel J. V., Leder P. Intervening sequence of DNA identified in the structural portion of a mouse beta-globin gene. Proc Natl Acad Sci U S A. 1978 Feb;75(2):725–729. doi: 10.1073/pnas.75.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tonegawa S., Maxam A. M., Tizard R., Bernard O., Gilbert W. Sequence of a mouse germ-line gene for a variable region of an immunoglobulin light chain. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1485–1489. doi: 10.1073/pnas.75.3.1485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Vogt V. M. Purification and further properties of single-strand-specific nuclease from Aspergillus oryzae. Eur J Biochem. 1973 Feb 15;33(1):192–200. doi: 10.1111/j.1432-1033.1973.tb02669.x. [DOI] [PubMed] [Google Scholar]
  27. Volckaert G., Van de Voorde A., Fiers W. Nucleotide sequence of the simian virus 40 small-t gene. Proc Natl Acad Sci U S A. 1978 May;75(5):2160–2164. doi: 10.1073/pnas.75.5.2160. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES