1887

Abstract

A recombinant baculovirus expressing the nucleocapsid gene (NP) of Newcastle disease virus (NDV), a member of the genus , has been generated and shown to express the native protein to high levels in insect cells. In contrast to the NP protein of the rubulavirus human parainfluenza virus 2, the NDV protein has been demonstrated by electron microscopy and caesium chloride gradient analysis to be capable of self-assembly to form nucleocapsid-like structures in the absence of other NDV proteins. These structures, which contained RNA that was resistant to micrococcal nuclease digestion, were also observed when the protein was expressed in , a phenomenon which was not inhibited by the presence of a 40 amino acid fusion region at the amino terminus of the protein. Further, the formation of these structures was inhibited by the co-expression of the phosphoprotein (P). Therefore, we conclude that the P protein acts as a chaperone, preventing uncontrolled encapsidation of non-viral RNA by NP protein.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-78-9-2335
1997-09-01
2024-04-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/78/9/9292023.html?itemId=/content/journal/jgv/10.1099/0022-1317-78-9-2335&mimeType=html&fmt=ahah

References

  1. Buchholz C. J., Spehner D., Drillien R., Neubert W. J., Homann H. E. 1993; The conserved N-terminal region of Sendai virus nucleocapsid protein NP is required for nucleocapsid assembly. Journal of Virology 67:5803–5812
    [Google Scholar]
  2. Chenik M., Chebli K., Gaudin Y., Blondel D. 1994; In vivo interaction of rabies virus phosphoprotein (P) and nucleoprotein (N): existence of two N-binding sites on P protein. Journal of General Virology 75:2889–2896
    [Google Scholar]
  3. Curran J., Marq J., Kolakofsky D. 1995a; An N-terminal domain of the Sendai paramyxovirus P protein acts as a chaperone for the NP protein during the nascent chain assembly step of genome replication. Journal of Virology 69:849–855
    [Google Scholar]
  4. Curran J., Boeck R., Lin-Marq N., Lupas A., Kolakofsky D. 1995b; Paramyxovirus phosphoproteins form homo-trimers as determined by an epitope dilution assay, via predicted coiled coils. Virology 214:139–149
    [Google Scholar]
  5. Errington W. 1996 Characterisation of the proteins involved in the transcription and replication of Newcastle disease virus PhD thesis University of Newcastle upon Tyne, UK:
    [Google Scholar]
  6. Errington W., Steward M. S., Emmerson P. T. 1995; A diagnostic immunoassay for Newcastle disease virus based on the nucleocapsid protein expressed by a recombinant baculovirus. Journal of Virological Methods 55:357–365
    [Google Scholar]
  7. Hamaguchi M., Yoshida T., Nishikawa K., Naruse H., Nagai Y. 1983; Transcriptive complex of Newcastle disease virus. I. Both P and L proteins are required to constitute and active complex. Virology 128:105–117
    [Google Scholar]
  8. Horikami S. M., Curran J., Kolakofsky D., Moyer S. A. 1992; Complexes of Sendai virus NP-P and P-L proteins are required for defective interfering genome replication in vitro. Journal of Virology 66:4901–4908
    [Google Scholar]
  9. Klug A. 1979; The assembly of tobacco mosaic virus : structure and specificity. The Harvey Lectures, Harvey Society of New York pp. 141–172 New York: Academic Press;
    [Google Scholar]
  10. Kurilla M. G., Stone H. O., Keene J. D. 1985; RNA sequence and translation properties of the 3′ end of the Newcastle disease virus genome. Virology 145:203–212
    [Google Scholar]
  11. Laemmli U. K. 1970; Cleavage of the structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  12. Lamb R. A., Kolakofsky D. 1996; Paramyxoviridae: the viruses and their replication. In Fields Virology, 3rd edn. pp. 1177–1204 Fields B. N., Knipe D. M., Howley P. M. Edited by Philadelphia: Lippincott-Raven;
    [Google Scholar]
  13. Madeley C. R., Field A. M. 1988 Virus Morphology, 2nd edn. Edinburgh and New York: Churchill Livingstone;
    [Google Scholar]
  14. Meric C., Spehner D., Mazarin V. 1994; Respiratory syncytial virus nucleocapsid protein (N) expressed in insect cells forms nucleocapsid-like structures. Virus Research 31:187–201
    [Google Scholar]
  15. Millar N. S., Emmerson P. T. 1988; Molecular cloning and nucleotide sequencing of Newcastle disease virus. In Developments in Veterinary Virology: Newcastle Disease pp. 79–97 Alexander D. J. Edited by Boston: Kluwer Academic Publishers;
    [Google Scholar]
  16. Mountcastle W. E., Compans R. W., Caliguiri L. A., Choppin P. W. 1970; Nucleocapsid protein subunits of simian virus 5, Newcastle disease virus and Sendai virus. Journal of Virology 6:677–684
    [Google Scholar]
  17. Murphy F. A., Fauquet C. M., Bishop D. H. L., Ghabrial S. A., Jarvis A. W., Martelli G. P., Mayo M. A., Summers M. D.editor 1995 Virus Taxonomy. Sixth Report of the International Committee on Taxonomy of Viruses Vienna & New York: Springer-Verlag;
    [Google Scholar]
  18. Nishio M., Tsurodome M., Kawano M., Watanabe N., Ohgimoto S., Ito M., Komada H., Ito Y. 1996; Interaction between the nucleocapsid protein (NP) and phosphoprotein (P) of human parainfluenza virus type 2 : one of the two NP binding sites on P is essential for granule formation. Journal of General Virology 77:2457–2463
    [Google Scholar]
  19. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  20. Spehner D., Kirn A., Drillier R. 1991; Assembly of nucleocapsid- like structures in animal cells infected with vaccinia virus recombinant encoding the measles virus nucleoprotein. Journal of Virology 65:6296–6300
    [Google Scholar]
  21. Steward M., Vipond I. B., Millar N. S., Emmerson P. T. 1993; RNA editing in Newcastle disease virus. Journal of General Virology 74:2539–2547
    [Google Scholar]
  22. Steward M. S., Samson A. C. R., Errington W., Emmerson P. T. 1995; The Newcastle disease virus V protein binds zinc. Archives of Virology 140:1321–1328
    [Google Scholar]
  23. Summers M. D., Smith G. E. 1987 A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures Texas Agricultural Experiment Station Bulletin 1555
    [Google Scholar]
  24. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets : procedure and some applications. Proceedings of the National Academy of Sciences, USA 76:4350–4354
    [Google Scholar]
  25. Warnes A., Fooks A. R., Dowesett A. B., Wilkinson G. W. G. 1995; Expression ofthe measles virus nucleocapsid protein in Escherichia coli and assembly of nucleocapsid-like structures. Gene 160:173–178
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-78-9-2335
Loading
/content/journal/jgv/10.1099/0022-1317-78-9-2335
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