1887

Abstract

RNA polymerase I transcription in transiently DNA-transfected cells has been used to express influenza virus vRNA molecules coding for chloramphenicol acetyltransferase (CAT) in an antisense orientation. Influenza virus superinfection provided viral RNA polymerase and other proteins required for transcriptional conversion of minus-strand vRNA into plus-strand viral mRNA molecules expressing CAT activity. This system has been used for analysis of the vRNA sequences which cooperatively constitute the vRNA promoter structure via nucleotide exchanges as well as deletions and insertions of both terminal segments. Several mutants caused greatly enhanced expression over wild-type levels, which was transmitted during serial passage of progeny virus. The data obtained for the mutations in various promoter elements support a model implicating double-stranded vRNA promoter structures in binding of viral polymerase, and in consecutive steps during initiation of RNA synthesis.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-76-7-1709
1995-07-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/76/7/JV0760071709.html?itemId=/content/journal/jgv/10.1099/0022-1317-76-7-1709&mimeType=html&fmt=ahah

References

  1. Baudin F., Bach C., Cusack S., Ruigrok R. W. H. 1994; Structure of influenza virus RNP. I. Influenza virus nucleoprotein melts secondary structure in panhandle RNA and exposes the bases to the solvent. EMBO Journal 13:3158–3165
    [Google Scholar]
  2. Braam J., Uhmanen I., Krug R. 1983; Molecular model of a eukaryotic transcription complex: functions and movements of influenza P proteins during capped RNA-primed transcription. Cell 34:609–618
    [Google Scholar]
  3. Compans R. W., Content J., Duesberg P. H. 1972; Structure of the ribonucleoprotein of influenza virus. Journal of Virology 10:795–800
    [Google Scholar]
  4. Fodor E., Seong B. L., Brownlee G. G. 1993; Photochemical cross-linking of influenza A polymerase to its virion RNA promoter defines a polymerase binding site at residues 9 to 12 of the promoter. Journal of General Virology 74:1327–1333
    [Google Scholar]
  5. Fodor E., Pritlove D. C., Brownlee G. G. 1994; The influenza virus panhandle is involved in the initiation of transcription. Journal of Virology 68:4092–4096
    [Google Scholar]
  6. Gorman M., Moffat L., Howard B. 1982; Recombinant genomes which express chloramphenicol acetyl-transferase in mammalian cells. Molecular and Cellular Biology 2:1044–1057
    [Google Scholar]
  7. Hagen M., Chung T. D. Y., Butcher J. A., Krystal M. 1994; Recombinant influenza virus polymerase: requirement of both 5′ and 3′ viral ends for endonuclease activity. Journal of Virology 68:1509–1515
    [Google Scholar]
  8. Honda A., Ueda K., Nagata K., Ishihama A. 1988; RNA polymerase of influenza virus. Role of NP on RNA chain elongation. Journal of Biochemistry 104:1021–1026
    [Google Scholar]
  9. Hsu M., Parvin J. D., Gupta S., Krystal M., Palese P. 1987; Genomic RNAs of influenza viruses are held in a circular conformation in virions and in infected cells by a terminal panhandle. Proceedings of the National Academy of Sciences, USA 84:8140–8144
    [Google Scholar]
  10. Li X., Palese P. 1992; Mutational analysis of the promoter required for influenza virus virion RNA synthesis. Journal of Virology 66:4331–4338
    [Google Scholar]
  11. Luo G., Luytjes W., Enami M., Palese P. 1991; The polyadenylation signal of influenza virus RNA involves a stretch of uridines followed by the RNA duplex of the panhandle structure. Journal of Virology 65:2861–2867
    [Google Scholar]
  12. Martin J., Albo C., Ortin J., Melero J. A., Portela A. 1992; In vitro reconstitution of active influenza virus ribonucleoprotein complexes using viral proteins purified from infected cells. Journal of General Virology 73:1855–1859
    [Google Scholar]
  13. Neumann G., Zobel A., Hobom G. 1994; RNA polymerase I-mediated expression of influenza viral RNA molecules. Virology 202:477–179
    [Google Scholar]
  14. Plotch S., Bouloy M., Krug R. M. 1979; Transfer of 5′-terminal cap of globin mRNA to influenza viral complementary RNA during transcription in vitro . Proceedings of the National Academy of Sciences, USA 76:1618–1622
    [Google Scholar]
  15. Seong B. L., Brownlee G. G. 1992; Nucleotides 9 to 11 of the influenza A virion RNA promoter are crucial for activity in vitro . Journal of General Virology 73:3115–3124
    [Google Scholar]
  16. Shapiro G., Krug R. 1988; Influenza virus RNA replication in vitro: synthesis of viral template RNAs and virion RNAs in the absence of an added primer. Journal of Virology 62:2285–2290
    [Google Scholar]
  17. Tiley L. S., Hagen M., Matthews J. T., Krystal M. 1994; Sequence-specific binding of influenza virus RNA polymerase to sequences located to the 5′ ends of the viral RNAs. Journal of Virology 68:5108–5116
    [Google Scholar]
  18. Zobel A., Neumann G., Hobom G. 1993; RNA polymerase I catalysed transcription of insert viral cDNA. Nucleic Acids Research 21:3607–3614
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-76-7-1709
Loading
/content/journal/jgv/10.1099/0022-1317-76-7-1709
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error