1887

Abstract

Both transcription and replication of the influenza virus RNA genome are catalysed by a virus-specific RNA polymerase. Recently, an assay, based on the synthesis of pppApG, for the initiation of replication by recombinant RNA polymerase in the absence of added primer was described. Here, these findings are extended to show that adenosine, AMP and ADP can each substitute for ATP in reactions catalysed by either recombinant ribonucleoprotein or RNA polymerase complexes with either model virion RNA (vRNA) or cRNA promoters. The use of either adenosine or AMP, rather than ATP, provides a convenient, sensitive and easy assay of replication initiation. Moreover, no pppApG was detected when a PB1–PA dimer, rather than the trimeric polymerase, was used to catalyse synthesis, contrasting with a previous report using baculovirus-expressed influenza RNA polymerase. Overall, it is suggested that the heterotrimeric polymerase is essential for the initiation of replication.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.82199-0
2006-11-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/87/11/3373.html?itemId=/content/journal/jgv/10.1099/vir.0.82199-0&mimeType=html&fmt=ahah

References

  1. Azzeh M., Flick R., Hobom G. 2001; Functional analysis of the influenza A virus cRNA promoter and construction of an ambisense transcription system. Virology 289:400–410 [CrossRef]
    [Google Scholar]
  2. Brosnan M. J., Chen L., Van Dyke T. A., Koretsky A. P. 1990; Free ADP levels in transgenic mouse liver expressing creatine kinase. J Biol Chem 265:20849–20855
    [Google Scholar]
  3. Brownlee G. G., Sharps J. L. 2002; The RNA polymerase of influenza A virus is stabilized by interaction with its viral RNA promoter. J Virol 76:7103–7113 [CrossRef]
    [Google Scholar]
  4. Crow M., Deng T., Addley M., Brownlee G. G. 2004; Mutational analysis of the influenza virus cRNA promoter and identification of nucleotides critical for replication. J Virol 78:6263–6270 [CrossRef]
    [Google Scholar]
  5. Deng T., Sharps J., Fodor E., Brownlee G. G. 2005; In vitro assembly of PB2 with a PB1-PA dimer supports a new model of assembly of influenza A virus polymerase subunits into a functional trimeric complex. J Virol 79:8669–8674 [CrossRef]
    [Google Scholar]
  6. Deng T., Vreede F. T., Brownlee G. G. 2006; Different de novo initiation strategies are used by influenza virus RNA polymerase on its cRNA and viral RNA promoters during viral RNA replication. J Virol 80:2337–2348 [CrossRef]
    [Google Scholar]
  7. Flick R., Hobom G. 1999; Interaction of influenza virus polymerase with viral RNA in the ‘corkscrew’ conformation. J Gen Virol 80:2565–2572
    [Google Scholar]
  8. Flick R., Neumann G., Hoffmann E., Neumeier E., Hobom G. 1996; Promoter elements in the influenza vRNA terminal structure. RNA 2:1046–1057
    [Google Scholar]
  9. Fodor E., Brownlee G. G. 2002; Influenza virus replication. In Influenza pp  1–30 Edited by Potter C. W. Amsterdam: Elsevier;
    [Google Scholar]
  10. Fodor E., Pritlove D. C., Brownlee G. G. 1994; The influenza virus panhandle is involved in the initiation of transcription. J Virol 68:4092–4096
    [Google Scholar]
  11. Honda A., Mizumoto K., Ishihama A. 2002; Minimum molecular architectures for transcription and replication of the influenza virus. Proc Natl Acad Sci U S A 99:13166–13171 [CrossRef]
    [Google Scholar]
  12. Kolpashchikov D. M., Honda A., Ishihama A. 2004; Structure-function relationship of the influenza virus RNA polymerase: primer-binding site on the PB1 subunit. Biochemistry 43:5882–5887 [CrossRef]
    [Google Scholar]
  13. Lamb R. A., Krug R. M. 2001; Orthomyxoviridae : the viruses and their replication. In Fields Virology pp  1487–1531 Edited by Knipe D. M., Howley P. M. Philadelphia, PA: Lippincott Williams & Wilkins;
    [Google Scholar]
  14. Lee M. T. M., Bishop K., Medcalf L., Elton D., Digard P., Tiley L. 2002; Definition of the minimal viral components required for the initiation of unprimed RNA synthesis by influenza virus RNA polymerase. Nucleic Acids Res 30:429–438 [CrossRef]
    [Google Scholar]
  15. Mills G. C., Schmalstieg F. C., Trimmer K. B., Goldman A. S., Goldblum R. M. 1976; Purine metabolism in adenosine deaminase deficiency. Proc Natl Acad Sci U S A 73:2867–2871 [CrossRef]
    [Google Scholar]
  16. Mullin A. E., Dalton R. M., Amorim M. J., Elton D., Digard P. 2004; Increased amounts of the influenza virus nucleoprotein do not promote higher levels of viral genome replication. J Gen Virol 85:3689–3698 [CrossRef]
    [Google Scholar]
  17. Neumann G., Brownlee G. G., Fodor E., Kawaoka Y. 2004; Orthomyxovirus replication, transcription, and polyadenylation. Curr Top Microbiol Immunol 283:121–143
    [Google Scholar]
  18. Pritlove D. C., Fodor E., Seong B. L., Brownlee G. G. 1995; In vitro transcription and polymerase binding studies of the termini of influenza A virus cRNA: evidence for a cRNA panhandle. J Gen Virol 76:2205–2213 [CrossRef]
    [Google Scholar]
  19. Puig O., Caspary F., Rigaut G., Rutz B., Bouveret E., Bragado-Nilsson E., Wilm M., Séraphin B. 2001; The tandem affinity purification (TAP) method: a general procedure of protein complex purification. Methods 24:218–229 [CrossRef]
    [Google Scholar]
  20. Randerath K., Randerath E. 1967; Ion-exchange thin-layer chromatography. XVII. Resolution of oligonucleotides from a pancreatic ribonuclease digest of RNA on PEI-cellulose. J Chromatogr 31:500–509 [CrossRef]
    [Google Scholar]
  21. Szymkowiak C., Kwan W. S., Su Q., Toner T. J., Shaw A. R., Youil R. 2003; Rapid method for the characterization of 3′ and 5′ UTRs of influenza viruses. J Virol Methods 107:15–20 [CrossRef]
    [Google Scholar]
  22. Vreede F. T., Jung T. E., Brownlee G. G. 2004; Model suggesting that replication of influenza virus is regulated by stabilization of replicative intermediates. J Virol 78:9568–9572 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.82199-0
Loading
/content/journal/jgv/10.1099/vir.0.82199-0
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