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. 1993 Jul;67(7):4358–4364. doi: 10.1128/jvi.67.7.4358-4364.1993

The hypervariable C-terminal tail of the Sendai paramyxovirus nucleocapsid protein is required for template function but not for RNA encapsidation.

J Curran 1, H Homann 1, C Buchholz 1, S Rochat 1, W Neubert 1, D Kolakofsky 1
PMCID: PMC237806  PMID: 8389932

Abstract

The paramyxovirus nucleocapsid proteins (NPs) are relatively well conserved, except for the C-terminal 20% (or ca. 100 amino acids), referred to as the tail. We have examined whether this hypervariable tail is required for genome synthesis, both in vitro, where synthesis is predominantly from the input templates, and in vivo, where multiple rounds of amplification occur. In these viruses, genome synthesis and assembly of the nascent chain are coupled. We find that the tail is required in vivo but not in vitro. Closer examination of the in vivo system showed that the tailless NP could encapsidate the genome chain but that amplification did not occur. We interpret these results as indicating that the tail is not required for RNA assembly but is required for the template to function in RNA synthesis. Relatively small deletions within the conserved N-terminal 80% of the protein, on the other hand, rendered the protein nonfunctional in either system. The possible functions of the tail in RNA synthesis are discussed.

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Selected References

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

  1. Blumberg B. M., Rose K., Simona M. G., Roux L., Giorgi C., Kolakofsky D. Analysis of the Sendai virus M gene and protein. J Virol. 1984 Nov;52(2):656–663. doi: 10.1128/jvi.52.2.656-663.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Buckland R., Giraudon P., Wild F. Expression of measles virus nucleoprotein in Escherichia coli: use of deletion mutants to locate the antigenic sites. J Gen Virol. 1989 Feb;70(Pt 2):435–441. doi: 10.1099/0022-1317-70-2-435. [DOI] [PubMed] [Google Scholar]
  3. Calain P., Curran J., Kolakofsky D., Roux L. Molecular cloning of natural paramyxovirus copy-back defective interfering RNAs and their expression from DNA. Virology. 1992 Nov;191(1):62–71. doi: 10.1016/0042-6822(92)90166-m. [DOI] [PubMed] [Google Scholar]
  4. Curran J., Boeck R., Kolakofsky D. The Sendai virus P gene expresses both an essential protein and an inhibitor of RNA synthesis by shuffling modules via mRNA editing. EMBO J. 1991 Oct;10(10):3079–3085. doi: 10.1002/j.1460-2075.1991.tb07860.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Curran J., Kolakofsky D. Scanning independent ribosomal initiation of the Sendai virus X protein. EMBO J. 1988 Sep;7(9):2869–2874. doi: 10.1002/j.1460-2075.1988.tb03143.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Curran J., Kolakofsky D. Scanning independent ribosomal initiation of the Sendai virus Y proteins in vitro and in vivo. EMBO J. 1989 Feb;8(2):521–526. doi: 10.1002/j.1460-2075.1989.tb03406.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Curran J., Marq J. B., Kolakofsky D. The Sendai virus nonstructural C proteins specifically inhibit viral mRNA synthesis. Virology. 1992 Aug;189(2):647–656. doi: 10.1016/0042-6822(92)90588-g. [DOI] [PubMed] [Google Scholar]
  8. Einberger H., Mertz R., Hofschneider P. H., Neubert W. J. Purification, renaturation, and reconstituted protein kinase activity of the Sendai virus large (L) protein: L protein phosphorylates the NP and P proteins in vitro. J Virol. 1990 Sep;64(9):4274–4280. doi: 10.1128/jvi.64.9.4274-4280.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Finch J. T., Gibbs A. J. Observations on the structure of the nucleocapsids of some paramyxoviruses. J Gen Virol. 1970 Jan;6(1):141–150. doi: 10.1099/0022-1317-6-1-141. [DOI] [PubMed] [Google Scholar]
  10. Fuerst T. R., Niles E. G., Studier F. W., Moss B. Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8122–8126. doi: 10.1073/pnas.83.21.8122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gill D. S., Takai S., Portner A., Kingsbury D. W. Mapping of antigenic domains of Sendai virus nucleocapsid protein expressed in Escherichia coli. J Virol. 1988 Dec;62(12):4805–4808. doi: 10.1128/jvi.62.12.4805-4808.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gotoh H., Shioda T., Sakai Y., Mizumoto K., Shibuta H. Rescue of Sendai virus from viral ribonucleoprotein-transfected cells by infection with recombinant vaccinia viruses carrying Sendai virus L and P/C genes. Virology. 1989 Aug;171(2):434–443. doi: 10.1016/0042-6822(89)90612-0. [DOI] [PubMed] [Google Scholar]
  13. Hamaguchi M., Yoshida T., Nishikawa K., Naruse H., Nagai Y. Transcriptive complex of Newcastle disease virus. I. Both L and P proteins are required to constitute an active complex. Virology. 1983 Jul 15;128(1):105–117. doi: 10.1016/0042-6822(83)90322-7. [DOI] [PubMed] [Google Scholar]
  14. Heggeness M. H., Scheid A., Choppin P. W. the relationship of conformational changes in the Sendai virus nucleocapsid to proteolytic cleavage of the NP polypeptide. Virology. 1981 Oct 30;114(2):555–562. doi: 10.1016/0042-6822(81)90235-x. [DOI] [PubMed] [Google Scholar]
  15. Homann H. E., Willenbrink W., Buchholz C. J., Neubert W. J. Sendai virus protein-protein interactions studied by a protein-blotting protein-overlay technique: mapping of domains on NP protein required for binding to P protein. J Virol. 1991 Mar;65(3):1304–1309. doi: 10.1128/jvi.65.3.1304-1309.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Horikami S. M., Curran J., Kolakofsky D., Moyer S. A. Complexes of Sendai virus NP-P and P-L proteins are required for defective interfering particle genome replication in vitro. J Virol. 1992 Aug;66(8):4901–4908. doi: 10.1128/jvi.66.8.4901-4908.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hsu C. H., Kingsbury D. W. Topography of phosphate residues in Sendai virus proteins. Virology. 1982 Jul 15;120(1):225–234. doi: 10.1016/0042-6822(82)90020-4. [DOI] [PubMed] [Google Scholar]
  18. Hudson L. D., Condra C., Lazzarini R. A. Cloning and expression of a viral phosphoprotein: structure suggests vesicular stomatitis virus NS may function by mimicking an RNA template. J Gen Virol. 1986 Aug;67(Pt 8):1571–1579. doi: 10.1099/0022-1317-67-8-1571. [DOI] [PubMed] [Google Scholar]
  19. Lamb R. A., Mahy B. W., Choppin P. W. The synthesis of sendai virus polypeptides in infected cells. Virology. 1976 Jan;69(1):116–131. doi: 10.1016/0042-6822(76)90199-9. [DOI] [PubMed] [Google Scholar]
  20. Lyn D., Gill D. S., Scroggs R. A., Portner A. The nucleoproteins of human parainfluenza virus type 1 and Sendai virus share amino acid sequences and antigenic and structural determinants. J Gen Virol. 1991 Apr;72(Pt 4):983–987. doi: 10.1099/0022-1317-72-4-983. [DOI] [PubMed] [Google Scholar]
  21. Lynch S., Kolakofsky D. Ends of the RNA within Sendai virus defective interfering nucleocapsids are not free. J Virol. 1978 Nov;28(2):584–589. doi: 10.1128/jvi.28.2.584-589.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Matsuoka Y., Ray R. Sequence analysis and expression of the human parainfluenza type 1 virus nucleoprotein gene. Virology. 1991 Mar;181(1):403–407. doi: 10.1016/0042-6822(91)90514-c. [DOI] [PubMed] [Google Scholar]
  23. Middleton Y., Tashiro M., Thai T., Oh J., Seymour J., Pritzer E., Klenk H. D., Rott R., Seto J. T. Nucleotide sequence analyses of the genes encoding the HN, M, NP, P, and L proteins of two host range mutants of Sendai virus. Virology. 1990 Jun;176(2):656–657. doi: 10.1016/0042-6822(90)90040-x. [DOI] [PubMed] [Google Scholar]
  24. Morgan E. M., Re G. G., Kingsbury D. W. Complete sequence of the Sendai virus NP gene from a cloned insert. Virology. 1984 May;135(1):279–287. doi: 10.1016/0042-6822(84)90137-5. [DOI] [PubMed] [Google Scholar]
  25. Mountcastle W. E., Compans R. W., Lackland H., Choppin P. W. Proteolytic cleavage of subunits of the nucleocapsid of the paramyxovirus simian virus 5. J Virol. 1974 Nov;14(5):1253–1261. doi: 10.1128/jvi.14.5.1253-1261.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Neubert W. J., Eckerskorn C., Homann H. E. Sendai virus NP gene codes for a 524 amino acid NP protein. Virus Genes. 1991 Jan;5(1):25–32. doi: 10.1007/BF00571728. [DOI] [PubMed] [Google Scholar]
  27. Parks G. D., Ward C. D., Lamb R. A. Molecular cloning of the NP and L genes of simian virus 5: identification of highly conserved domains in paramyxovirus NP and L proteins. Virus Res. 1992 Mar;22(3):259–279. doi: 10.1016/0168-1702(92)90057-g. [DOI] [PubMed] [Google Scholar]
  28. Perrault J., Clinton G. M., McClure M. A. RNP template of vesicular stomatitis virus regulates transcription and replication functions. Cell. 1983 Nov;35(1):175–185. doi: 10.1016/0092-8674(83)90220-9. [DOI] [PubMed] [Google Scholar]
  29. Shioda T., Hidaka Y., Kanda T., Shibuta H., Nomoto A., Iwasaki K. Sequence of 3,687 nucleotides from the 3' end of Sendai virus genome RNA and the predicted amino acid sequences of viral NP, P and C proteins. Nucleic Acids Res. 1983 Nov 11;11(21):7317–7330. doi: 10.1093/nar/11.21.7317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Vidal S., Kolakofsky D. Modified model for the switch from Sendai virus transcription to replication. J Virol. 1989 May;63(5):1951–1958. doi: 10.1128/jvi.63.5.1951-1958.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yuasa T., Bando H., Kawano M., Tsurudome M., Nishio M., Kondo K., Komada H., Ito Y. Sequence analyses of the 3' genome end and NP gene of human parainfluenza type 2 virus: sequence variation of the gene-starting signal and the conserved 3' end. Virology. 1990 Dec;179(2):777–784. doi: 10.1016/0042-6822(90)90145-h. [DOI] [PubMed] [Google Scholar]

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