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. 1995 Feb;69(2):849–855. doi: 10.1128/jvi.69.2.849-855.1995

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.

J Curran 1, J B Marq 1, D Kolakofsky 1
PMCID: PMC188651  PMID: 7815552

Abstract

Two domains involved in RNA synthesis have recently been found within the N-terminal 77 amino acids of the Sendai virus P protein. One domain is required for RNA synthesis per se and has properties in common with the transactivation domains of cellular transcription factors. The second domain is thought to be specifically required for the nascent chain assembly step in genome replication. We have further mapped this second domain by the construction of chimeric and deleted P proteins to amino acids 33 to 41 of P and by examining the abilities of these P proteins to support DI genome replication in vivo. Using glycerol gradient sedimentation, we have shown that this domain is required to form a stable complex with unassembled NP (P-NP0) and to prevent NP from assembling illegitimately, i.e., independently of the concurrent assembly of a nascent viral genome. Since the P-NP0 complex represents the functional form of unassembled NP which is delivered to the nascent chain during genome replication, and since amino acids 33 to 41 are not required for the stable interaction of P with the assembled NP of the nucleocapsid, this chaperone function of P is not required for mRNA synthesis or the RNA synthesis step of genome replication.

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Selected References

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  1. Buchholz C. J., Spehner D., Drillien R., Neubert W. J., Homann H. E. The conserved N-terminal region of Sendai virus nucleocapsid protein NP is required for nucleocapsid assembly. J Virol. 1993 Oct;67(10):5803–5812. doi: 10.1128/jvi.67.10.5803-5812.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Curran J., Homann H., Buchholz C., Rochat S., Neubert W., Kolakofsky D. The hypervariable C-terminal tail of the Sendai paramyxovirus nucleocapsid protein is required for template function but not for RNA encapsidation. J Virol. 1993 Jul;67(7):4358–4364. doi: 10.1128/jvi.67.7.4358-4364.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Curran J., Pelet T., Kolakofsky D. An acidic activation-like domain of the Sendai virus P protein is required for RNA synthesis and encapsidation. Virology. 1994 Aug 1;202(2):875–884. doi: 10.1006/viro.1994.1409. [DOI] [PubMed] [Google Scholar]
  6. Deshpande K. L., Portner A. Monoclonal antibodies to the P protein of Sendai virus define its structure and role in transcription. Virology. 1985 Jan 15;140(1):125–134. doi: 10.1016/0042-6822(85)90451-9. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Field J., Nikawa J., Broek D., MacDonald B., Rodgers L., Wilson I. A., Lerner R. A., Wigler M. Purification of a RAS-responsive adenylyl cyclase complex from Saccharomyces cerevisiae by use of an epitope addition method. Mol Cell Biol. 1988 May;8(5):2159–2165. doi: 10.1128/mcb.8.5.2159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. 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]
  12. Howard M., Wertz G. Vesicular stomatitis virus RNA replication: a role for the NS protein. J Gen Virol. 1989 Oct;70(Pt 10):2683–2694. doi: 10.1099/0022-1317-70-10-2683. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Ma J., Ptashne M. Deletion analysis of GAL4 defines two transcriptional activating segments. Cell. 1987 Mar 13;48(5):847–853. doi: 10.1016/0092-8674(87)90081-x. [DOI] [PubMed] [Google Scholar]
  15. Matsuoka Y., Curran J., Pelet T., Kolakofsky D., Ray R., Compans R. W. The P gene of human parainfluenza virus type 1 encodes P and C proteins but not a cysteine-rich V protein. J Virol. 1991 Jun;65(6):3406–3410. doi: 10.1128/jvi.65.6.3406-3410.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mikaelian I., Sergeant A. A general and fast method to generate multiple site directed mutations. Nucleic Acids Res. 1992 Jan 25;20(2):376–376. doi: 10.1093/nar/20.2.376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mottet G., Curran J., Roux L. Intracellular stability of nonreplicating paramyxovirus nucleocapsids. Virology. 1990 May;176(1):1–7. doi: 10.1016/0042-6822(90)90223-e. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Orvell C., Grandien M. The effects of monoclonal antibodies on biologic activities of structural proteins of Sendai virus. J Immunol. 1982 Dec;129(6):2779–2787. [PubMed] [Google Scholar]
  20. Pelet T., Curran J., Kolakofsky D. The P gene of bovine parainfluenza virus 3 expresses all three reading frames from a single mRNA editing site. EMBO J. 1991 Feb;10(2):443–448. doi: 10.1002/j.1460-2075.1991.tb07966.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Peluso R. W., Moyer S. A. Viral proteins required for the in vitro replication of vesicular stomatitis virus defective interfering particle genome RNA. Virology. 1988 Feb;162(2):369–376. doi: 10.1016/0042-6822(88)90477-1. [DOI] [PubMed] [Google Scholar]
  22. Rose J. K., Buonocore L., Whitt M. A. A new cationic liposome reagent mediating nearly quantitative transfection of animal cells. Biotechniques. 1991 Apr;10(4):520–525. [PubMed] [Google Scholar]
  23. Ryan K. W., Morgan E. M., Portner A. Two noncontiguous regions of Sendai virus P protein combine to form a single nucleocapsid binding domain. Virology. 1991 Jan;180(1):126–134. doi: 10.1016/0042-6822(91)90016-5. [DOI] [PubMed] [Google Scholar]
  24. Ryan K. W., Portner A., Murti K. G. Antibodies to paramyxovirus nucleoproteins define regions important for immunogenicity and nucleocapsid assembly. Virology. 1993 Mar;193(1):376–384. doi: 10.1006/viro.1993.1134. [DOI] [PubMed] [Google Scholar]
  25. Ryan K. W., Portner A. Separate domains of Sendai virus P protein are required for binding to viral nucleocapsids. Virology. 1990 Feb;174(2):515–521. doi: 10.1016/0042-6822(90)90105-z. [DOI] [PubMed] [Google Scholar]
  26. Sakai Y., Suzu S., Shioda T., Shibuta H. Nucleotide sequence of the bovine parainfluenza 3 virus genome: its 3' end and the genes of NP, P, C and M proteins. Nucleic Acids Res. 1987 Apr 10;15(7):2927–2944. doi: 10.1093/nar/15.7.2927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Smallwood S., Ryan K. W., Moyer S. A. Deletion analysis defines a carboxyl-proximal region of Sendai virus P protein that binds to the polymerase L protein. Virology. 1994 Jul;202(1):154–163. doi: 10.1006/viro.1994.1331. [DOI] [PubMed] [Google Scholar]
  28. Spehner D., Kirn A., Drillien R. Assembly of nucleocapsidlike structures in animal cells infected with a vaccinia virus recombinant encoding the measles virus nucleoprotein. J Virol. 1991 Nov;65(11):6296–6300. doi: 10.1128/jvi.65.11.6296-6300.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Vidal S., Curran J., Kolakofsky D. Editing of the Sendai virus P/C mRNA by G insertion occurs during mRNA synthesis via a virus-encoded activity. J Virol. 1990 Jan;64(1):239–246. doi: 10.1128/jvi.64.1.239-246.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Vidal S., Curran J., Orvell C., Kolakofsky D. Mapping of monoclonal antibodies to the Sendai virus P protein and the location of its phosphates. J Virol. 1988 Jun;62(6):2200–2203. doi: 10.1128/jvi.62.6.2200-2203.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]

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