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. 1989 Dec;63(12):5142–5152. doi: 10.1128/jvi.63.12.5142-5152.1989

Promoter analysis of influenza virus RNA polymerase.

J D Parvin 1, P Palese 1, A Honda 1, A Ishihama 1, M Krystal 1
PMCID: PMC251177  PMID: 2585601

Abstract

Influenza virus polymerase, which was prepared depleted of viral RNA, was used to copy small RNA templates prepared from plasmid-encoded sequences. Template constructions containing only the 3' end of genomic RNA were shown to be efficiently copied, indicating that the promoter lay solely within the 15-nucleotide 3' terminus. Sequences not specific for the influenza virus termini were not copied, and, surprisingly, RNAs containing termini identical to those from plus-sense cRNA were copied at low levels. The specificity for recognition of the virus sense promoter was further defined by site-specific mutagenesis. It was also found that increased levels of viral protein were required in order to catalyze both the cap endonuclease-primed and primer-free RNA synthesis from these model templates, as well as from genomic-length RNAs. This finding indicates that the reconstituted system has catalytic properties very similar to those of native viral ribonucleoprotein complexes.

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Selected References

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

  1. Beaton A. R., Krug R. M. Transcription antitermination during influenza viral template RNA synthesis requires the nucleocapsid protein and the absence of a 5' capped end. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6282–6286. doi: 10.1073/pnas.83.17.6282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bishop D. H., Obijeski J. F., Simpson R. W. Transcription of the influenza ribonucleic acid genome by a virion polymerase. I. Optimal conditions for in vitro activity of the ribonucleic acid-dependent ribonucleic acid polymerase. J Virol. 1971 Jul;8(1):66–73. doi: 10.1128/jvi.8.1.66-73.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bouloy M., Plotch S. J., Krug R. M. Both the 7-methyl and the 2'-O-methyl groups in the cap of mRNA strongly influence its ability to act as primer for influenza virus RNA transcription. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3952–3956. doi: 10.1073/pnas.77.7.3952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dreher T. W., Bujarski J. J., Hall T. C. Mutant viral RNAs synthesized in vitro show altered aminoacylation and replicase template activities. Nature. 1984 Sep 13;311(5982):171–175. doi: 10.1038/311171a0. [DOI] [PubMed] [Google Scholar]
  5. Dreher T. W., Hall T. C. Mutational analysis of the sequence and structural requirements in brome mosaic virus RNA for minus strand promoter activity. J Mol Biol. 1988 May 5;201(1):31–40. doi: 10.1016/0022-2836(88)90436-6. [DOI] [PubMed] [Google Scholar]
  6. Emerson S. U., Yu Y. Both NS and L proteins are required for in vitro RNA synthesis by vesicular stomatitis virus. J Virol. 1975 Jun;15(6):1348–1356. doi: 10.1128/jvi.15.6.1348-1356.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hay A. J., Lomniczi B., Bellamy A. R., Skehel J. J. Transcription of the influenza virus genome. Virology. 1977 Dec;83(2):337–355. doi: 10.1016/0042-6822(77)90179-9. [DOI] [PubMed] [Google Scholar]
  8. Honda A., Uéda K., Nagata K., Ishihama A. Identification of the RNA polymerase-binding site on genome RNA of influenza virus. J Biochem. 1987 Nov;102(5):1241–1249. doi: 10.1093/oxfordjournals.jbchem.a122163. [DOI] [PubMed] [Google Scholar]
  9. Honda A., Uéda K., Nagata K., Ishihama A. RNA polymerase of influenza virus: role of NP in RNA chain elongation. J Biochem. 1988 Dec;104(6):1021–1026. doi: 10.1093/oxfordjournals.jbchem.a122569. [DOI] [PubMed] [Google Scholar]
  10. Hsu M. T., Parvin J. D., Gupta S., Krystal M., Palese P. Genomic RNAs of influenza viruses are held in a circular conformation in virions and in infected cells by a terminal panhandle. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8140–8144. doi: 10.1073/pnas.84.22.8140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ishihama A., Nagata K. Viral RNA polymerases. CRC Crit Rev Biochem. 1988;23(1):27–76. doi: 10.3109/10409238809103119. [DOI] [PubMed] [Google Scholar]
  12. Kaplan G., Lubinski J., Dasgupta A., Racaniello V. R. In vitro synthesis of infectious poliovirus RNA. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8424–8428. doi: 10.1073/pnas.82.24.8424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kato A., Mizumoto K., Ishihama A. Purification and enzymatic properties of an RNA polymerase-RNA complex from influenza virus. Virus Res. 1985 Sep;3(2):115–127. doi: 10.1016/0168-1702(85)90002-4. [DOI] [PubMed] [Google Scholar]
  14. Kingsbury D. W., Jones I. M., Murti K. G. Assembly of influenza ribonucleoprotein in vitro using recombinant nucleoprotein. Virology. 1987 Feb;156(2):396–403. doi: 10.1016/0042-6822(87)90419-3. [DOI] [PubMed] [Google Scholar]
  15. Levis R., Weiss B. G., Tsiang M., Huang H., Schlesinger S. Deletion mapping of Sindbis virus DI RNAs derived from cDNAs defines the sequences essential for replication and packaging. Cell. 1986 Jan 17;44(1):137–145. doi: 10.1016/0092-8674(86)90492-7. [DOI] [PubMed] [Google Scholar]
  16. Mirakhur B., Peluso R. W. In vitro assembly of a functional nucleocapsid from the negative-stranded genome RNA of a defective interfering particle of vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7511–7515. doi: 10.1073/pnas.85.20.7511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Naito S., Ishihama A. Function and structure of RNA polymerase from vesicular stomatitis virus. J Biol Chem. 1976 Jul 25;251(14):4307–4314. [PubMed] [Google Scholar]
  18. Plotch S. J., Bouloy M., Ulmanen I., Krug R. M. A unique cap(m7GpppXm)-dependent influenza virion endonuclease cleaves capped RNAs to generate the primers that initiate viral RNA transcription. Cell. 1981 Mar;23(3):847–858. doi: 10.1016/0092-8674(81)90449-9. [DOI] [PubMed] [Google Scholar]
  19. Robertson J. S., Schubert M., Lazzarini R. A. Polyadenylation sites for influenza virus mRNA. J Virol. 1981 Apr;38(1):157–163. doi: 10.1128/jvi.38.1.157-163.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rochovansky O. M. RNA synthesis by ribonucleoprotein-polymerase complexes isolated from influenza virus. Virology. 1976 Sep;73(2):327–338. doi: 10.1016/0042-6822(76)90394-9. [DOI] [PubMed] [Google Scholar]
  21. Scholtissek C., Becht H. Binding of ribonucleic acids to the RNP-antigen protein of influenza viruses. J Gen Virol. 1971 Jan;10(1):11–16. doi: 10.1099/0022-1317-10-1-11. [DOI] [PubMed] [Google Scholar]
  22. Shapiro G. I., Krug R. M. Influenza virus RNA replication in vitro: synthesis of viral template RNAs and virion RNAs in the absence of an added primer. J Virol. 1988 Jul;62(7):2285–2290. doi: 10.1128/jvi.62.7.2285-2290.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Studier F. W., Dunn J. J. Organization and expression of bacteriophage T7 DNA. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 2):999–1007. doi: 10.1101/sqb.1983.047.01.114. [DOI] [PubMed] [Google Scholar]
  24. Szewczyk B., Laver W. G., Summers D. F. Purification, thioredoxin renaturation, and reconstituted activity of the three subunits of the influenza A virus RNA polymerase. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7907–7911. doi: 10.1073/pnas.85.21.7907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Takeuchi K., Nagata K., Ishihama A. In vitro synthesis of influenza viral RNA: characterization of an isolated nuclear system that supports transcription of influenza viral RNA. J Biochem. 1987 Apr;101(4):837–845. doi: 10.1093/oxfordjournals.jbchem.a121950. [DOI] [PubMed] [Google Scholar]
  26. Ulmanen I., Broni B., Krug R. M. Influenza virus temperature-sensitive cap (m7GpppNm)-dependent endonuclease. J Virol. 1983 Jan;45(1):27–35. doi: 10.1128/jvi.45.1.27-35.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ward C. D., Stokes M. A., Flanegan J. B. Direct measurement of the poliovirus RNA polymerase error frequency in vitro. J Virol. 1988 Feb;62(2):558–562. doi: 10.1128/jvi.62.2.558-562.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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