Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1983 Oct 25;11(20):7031–7042. doi: 10.1093/nar/11.20.7031

Initiation of RNA synthesis in vitro by vesicular stomatitis virus: single internal initiation in the presence of aurintricarboxylic acid and vanadyl ribonucleoside complexes.

S Talib, J E Hearst
PMCID: PMC326436  PMID: 6314272

Abstract

In the presence of aurintricarboxylic acid (ATA) and vanadyl ribonucleoside complexes (VRC), we have isolated and characterized a small RNA, product of VSV in vitro transcription. This RNA is capped and lacks poly(A) at its 3'-end. Nucleotide sequence analysis revealed that this RNA corresponds to the 5'-terminal transcription product of the N-gene. The termination of the transcription occurs precisely at the 118th base from the 3'-end of the VSV genome. Analysis of the nucleotide sequence around this region reveals a potential secondary structure. Photoreaction of the VSV with 4'-substituted psoralen fails to inhibit the synthesis of the 68-mer RNA under conditions where full length mRNA synthesis is blocked, indicating that the psoralen binding site is located further into the N-gene. Since this RNA is the only in vitro transcription product synthesized under these conditions, the existence of two types of polymerase activities, one for the synthesis of leader RNA and one for mRNA, is suggested.

Full text

PDF
7035

Images in this article

Selected References

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

  1. Abraham G., Rhodes D. P., Banerjee A. K. The 5' terminal structure of the methylated mRNA synthesized in vitro by vesicular stomatitis virus. Cell. 1975 May;5(1):51–58. doi: 10.1016/0092-8674(75)90091-4. [DOI] [PubMed] [Google Scholar]
  2. Ball L. A., White C. N. Order of transcription of genes of vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1976 Feb;73(2):442–446. doi: 10.1073/pnas.73.2.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Donis-Keller H., Maxam A. M., Gilbert W. Mapping adenines, guanines, and pyrimidines in RNA. Nucleic Acids Res. 1977 Aug;4(8):2527–2538. doi: 10.1093/nar/4.8.2527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Emerson S. U. Reconstitution studies detect a single polymerase entry site on the vesicular stomatitis virus genome. Cell. 1982 Dec;31(3 Pt 2):635–642. doi: 10.1016/0092-8674(82)90319-1. [DOI] [PubMed] [Google Scholar]
  5. Gallione C. J., Greene J. R., Iverson L. E., Rose J. K. Nucleotide sequences of the mRNA's encoding the vesicular stomatitis virus N and NS proteins. J Virol. 1981 Aug;39(2):529–535. doi: 10.1128/jvi.39.2.529-535.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hay N., Skolnik-David H., Aloni Y. Attenuation in the control of SV40 gene expression. Cell. 1982 May;29(1):183–193. doi: 10.1016/0092-8674(82)90102-7. [DOI] [PubMed] [Google Scholar]
  7. Herman R. C., Adler S., Lazzarini R. A., Colonno R. J., Banerjee A. K., Westphal H. Intervening polyadenylate sequences in RNA transcripts of vesicular stomatitis virus. Cell. 1978 Oct;15(2):587–596. doi: 10.1016/0092-8674(78)90027-2. [DOI] [PubMed] [Google Scholar]
  8. Herman R. C., Schubert M., Keene J. D., Lazzarini R. A. Polycistronic vesicular stomatitis virus RNA transcripts. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4662–4665. doi: 10.1073/pnas.77.8.4662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Iverson L. E., Rose J. K. Sequential synthesis of 5'-proximal vesicular stomatitis virus mRNA sequences. J Virol. 1982 Oct;44(1):356–365. doi: 10.1128/jvi.44.1.356-365.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kaufmann G., Choder M., Groner Y. Synthesis of carrier-free beta-32P-nucleosides-triphosphate in almost quantitative yields. Anal Biochem. 1980 Nov 15;109(1):198–202. doi: 10.1016/0003-2697(80)90029-9. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Pinney D. F., Emerson S. U. Identification and characterization of a group of discrete initiated oligonucleotides transcribed in vitro from the 3' terminus of the N-gene of vesicular stomatitis virus. J Virol. 1982 Jun;42(3):889–896. doi: 10.1128/jvi.42.3.889-896.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pinney D. F., Emerson S. U. In vitro synthesis of triphosphate-initiated N-gene mRNA oligonucleotides is regulated by the matrix protein of vesicular stomatitis virus. J Virol. 1982 Jun;42(3):897–904. doi: 10.1128/jvi.42.3.897-904.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Platt T. Termination of transcription and its regulation in the tryptophan operon of E. coli. Cell. 1981 Apr;24(1):10–23. doi: 10.1016/0092-8674(81)90496-7. [DOI] [PubMed] [Google Scholar]
  16. Rose J. K. Heterogneeous 5'-terminal structures occur on vesicular stomatitis virus mRNAs. J Biol Chem. 1975 Oct 25;250(20):8098–8104. [PubMed] [Google Scholar]
  17. Roy P., Bishop D. H. Genome homology of vesicular stomatitis virus and defective T particles and evidence for the sequential transcription of the virion ribonucleic acid. J Virol. 1972 Jun;9(6):946–955. doi: 10.1128/jvi.9.6.946-955.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schubert M., Harmison G. G., Sprague J., Condra C. S., Lazzarini R. A. In vitro transcription of vesicular stomatitis virus: initiation with GTP at a specific site within the N cistron. J Virol. 1982 Jul;43(1):166–173. doi: 10.1128/jvi.43.1.166-173.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schulz-Harder B., Tata J. R. Inhibition of nuclear ribonuclease activity during transcription in vitro by aurintricarboxylic acid and vanadyl riboncleoside complexes. Biochem Biophys Res Commun. 1982 Feb 11;104(3):903–910. doi: 10.1016/0006-291x(82)91334-1. [DOI] [PubMed] [Google Scholar]
  20. Singh B., Thornton G. B., Roy J., Talib S., De B. P., Banerjee A. K. Frequent intragenic transcription termination within the N gene of vesicular stomatitis virus. Virology. 1982 Oct 30;122(2):239–250. doi: 10.1016/0042-6822(82)90224-0. [DOI] [PubMed] [Google Scholar]
  21. Talib S., Banerjee A. K. Covalent attachment of psoralen to a single site on vesicular stomatitis virus genome RNA blocks expression of viral genes. Virology. 1982 Apr 30;118(2):430–438. doi: 10.1016/0042-6822(82)90362-2. [DOI] [PubMed] [Google Scholar]
  22. Talib S., Banerjee A. K. Protamine--a potent inhibitor of vesicular stomatitis virus transcriptase in vitro. Biochem Biophys Res Commun. 1981 Feb 12;98(3):875–883. doi: 10.1016/0006-291x(81)91192-x. [DOI] [PubMed] [Google Scholar]
  23. 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]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES