1887

Abstract

SUMMARY

Soluble transcriptase containing the L and the NS proteins was isolated from purified vesicular stomatitis virus and its binding with the template ribonucleoprotein containing the N protein-RNA complex was studied with respect to its ability to initiate and synthesize RNA . By using u.v.-irradiated template reconstituted with soluble transcriptase, it was shown that the synthesis of leader RNA and other small initiated mRNA sequences continued while full-length mRNA synthesis decreased by 90%. In the presence of ATP and CTP, the reconstituted complex synthesized polyphosphorylated oligonucleotides which include AC, AAC and AACA which represent 5′-terminal sequences transcribed from the leader template and genes coding for mRNAs. In the presence of arabinosyl ATP, an inhibitor of RNA synthesis , the synthesis of leader RNA was found to be inhibited considerably more than other small initiated mRNA sequences. Reconstitution of RNA synthesis with soluble transcriptase and template in the presence of viral matrix (M) protein at low ionic condition resulted in virtual cessation of leader RNA synthesis, although the synthesis of small initiated N mRNA, 11 to 14 bases, continued. These results suggest that transcriptase can bind at multiple sites on the genome template and initiate RNA chains.

Keyword(s): RNA synthesis , transcriptase and VSV
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/content/journal/jgv/10.1099/0022-1317-65-3-663
1984-03-01
2024-04-19
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References

  1. Abraham G., Banerjee A. K. 1976a; Sequential transcription of the genes of vesicular stomatitis virus. Proceedings of the National Academy of Sciences, U,. S,. A 73:1504–1508
    [Google Scholar]
  2. Abraham G., BAnerjee A. K. 1976b; The nature of the RNA products synthesized in vitro by subviral components of vesicular stomatitis virus. Virology 71:230–241
    [Google Scholar]
  3. Ball L. A., White C. N. 1976; Order of transcription of genes of vesicular stomatitis virus. Proceedings of the National Academy of Sciences, U,. S,. A 73:442–446
    [Google Scholar]
  4. Banerjee A. K., Abraham G., Colonno R. J. 1977; Vesicular stomatitis virus: mode of transcription. Journal of General Virology 34:1–8
    [Google Scholar]
  5. Carroll A. R., Wagner R. R. 1979; Role of the membrane (M) protein in endogenous inhibition of in vitro transcription by vesicular stomatitis virus. Journal of Virology 29:134–142
    [Google Scholar]
  6. Chanda P. K., Banerjee A. K. 1980; Inhibition of vesicular stomatitis virus transcriptase in vitro by phosphonoformate and ara-ATP. Virology 107:562–566
    [Google Scholar]
  7. Chanda P. K., Banerjee A. K. 1981; Identification of promoter proximal oligonucleotides and a unique dinucleotide, pppGpC, from in vitro transcription products of vesicular stomatitis virus. Journal of Virology 39:93–103
    [Google Scholar]
  8. De B. P., Thornton G. B., Luk D., Banerjee A. K. 1982; Purified matrix protein of vesicular stomatitis virus blocks viral transcription in vitro. Proceedings of the National Academy of Sciences, U. S. A 79:7137–7141
    [Google Scholar]
  9. Emerson S. U. 1982; Reconstitution studies detect a single polymerase entry site on the vesicular stomatitis virus genome. Cell 31:635–642
    [Google Scholar]
  10. Emerson S. U., Yu Y. H. 1975; Both NS and L proteins are required for in vitro RNA synthesis by vesicular stomatitis virus. Journal of Virology 15:1348–1356
    [Google Scholar]
  11. Hsu C. H., Kingsbury D. W. 1982; NS phosphoprotein of vesicular stomatitis virus: subspecies separated by electrophoresis and isoelectric focusing. Journal of Virology 42:342–345
    [Google Scholar]
  12. Kingsford L., Emerson S. U. 1980; Transcriptional activities of different phosphorylated species of NS protein purified from vesicular stomatitis virus and cytoplasm of infected cells. Journal of Virology 33:1097–1105
    [Google Scholar]
  13. Pinney D. F., Emerson S. U. 1982a; 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. Journal of Virology 42:889–896
    [Google Scholar]
  14. Pinney D. F., Emerson S. U. 1982b; In vitro synthesis of triphosphate initiated N-gene mRNA oligonucleotides is regulated by the matrix protein of vesicular stomatitis virus. Journal of Virology 42:897–904
    [Google Scholar]
  15. Schubert M., Harmison G. G., Sprague J., Condra C. S., Lazzarini R. A. 1982; In vitro transcription of vesicular stomatitis virus: initiation with GTP at a specific site within the N cistron. Journal of Virology 43:166–173
    [Google Scholar]
  16. Talib S., Banerjee A. K. 1982; Covalent attachment of psoralen to a single site on vesicular stomatitis virus genome RNA blocks expression of viral genes. Virology 118:430–438
    [Google Scholar]
  17. Testa D., Chanda P. K., Banerjee A. K. 1980; Unique mode of transcription in vitro by vesicular stomatitis virus. Cell 21:267–275
    [Google Scholar]
  18. Wilson T., Lenard J. 1981; Interaction of wild-type and mutant M protein of vesicular stomatitis virus with nucleocapsids in vitro. Biochemistry 20:1349–1354
    [Google Scholar]
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