Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1995 Dec;69(12):8123–8126. doi: 10.1128/jvi.69.12.8123-8126.1995

Posttranslational signal peptidase cleavage at the flavivirus C-prM junction in vitro.

C E Stocks 1, M Lobigs 1
PMCID: PMC189766  PMID: 7494334

Abstract

We have investigated the cleavages at the flavivirus capsid-prM protein junction in vitro. When expressed in the absence of the flavivirus proteinase, capsid and prM, which are separated by an internal signal sequence, exist as a membrane-spanning precursor protein. Here we show the induction of posttranslational signal peptidase cleavage of prM by trypsin cleavage of a cytoplasmic region of this precursor protein.

Full Text

The Full Text of this article is available as a PDF (194.1 KB).

Selected References

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

  1. Amberg S. M., Nestorowicz A., McCourt D. W., Rice C. M. NS2B-3 proteinase-mediated processing in the yellow fever virus structural region: in vitro and in vivo studies. J Virol. 1994 Jun;68(6):3794–3802. doi: 10.1128/jvi.68.6.3794-3802.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bruss V., Lu X., Thomssen R., Gerlich W. H. Post-translational alterations in transmembrane topology of the hepatitis B virus large envelope protein. EMBO J. 1994 May 15;13(10):2273–2279. doi: 10.1002/j.1460-2075.1994.tb06509.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chambers T. J., Grakoui A., Rice C. M. Processing of the yellow fever virus nonstructural polyprotein: a catalytically active NS3 proteinase domain and NS2B are required for cleavages at dibasic sites. J Virol. 1991 Nov;65(11):6042–6050. doi: 10.1128/jvi.65.11.6042-6050.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chambers T. J., Hahn C. S., Galler R., Rice C. M. Flavivirus genome organization, expression, and replication. Annu Rev Microbiol. 1990;44:649–688. doi: 10.1146/annurev.mi.44.100190.003245. [DOI] [PubMed] [Google Scholar]
  5. Chambers T. J., McCourt D. W., Rice C. M. Production of yellow fever virus proteins in infected cells: identification of discrete polyprotein species and analysis of cleavage kinetics using region-specific polyclonal antisera. Virology. 1990 Jul;177(1):159–174. doi: 10.1016/0042-6822(90)90470-c. [DOI] [PubMed] [Google Scholar]
  6. Chambers T. J., Weir R. C., Grakoui A., McCourt D. W., Bazan J. F., Fletterick R. J., Rice C. M. Evidence that the N-terminal domain of nonstructural protein NS3 from yellow fever virus is a serine protease responsible for site-specific cleavages in the viral polyprotein. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8898–8902. doi: 10.1073/pnas.87.22.8898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dalgarno L., Trent D. W., Strauss J. H., Rice C. M. Partial nucleotide sequence of the Murray Valley encephalitis virus genome. Comparison of the encoded polypeptides with yellow fever virus structural and non-structural proteins. J Mol Biol. 1986 Feb 5;187(3):309–323. doi: 10.1016/0022-2836(86)90435-3. [DOI] [PubMed] [Google Scholar]
  8. Falgout B., Pethel M., Zhang Y. M., Lai C. J. Both nonstructural proteins NS2B and NS3 are required for the proteolytic processing of dengue virus nonstructural proteins. J Virol. 1991 May;65(5):2467–2475. doi: 10.1128/jvi.65.5.2467-2475.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gruenberg A., Wright P. J. Processing of dengue virus type 2 structural proteins containing deletions in hydrophobic domains. Arch Virol. 1992;122(1-2):77–94. doi: 10.1007/BF01321119. [DOI] [PubMed] [Google Scholar]
  10. Hall R. A., Kay B. H., Burgess G. W., Clancy P., Fanning I. D. Epitope analysis of the envelope and non-structural glycoproteins of Murray Valley encephalitis virus. J Gen Virol. 1990 Dec;71(Pt 12):2923–2930. doi: 10.1099/0022-1317-71-12-2923. [DOI] [PubMed] [Google Scholar]
  11. Hegner M., von Kieckebusch-Gück A., Falchetto R., James P., Semenza G., Mantei N. Single amino acid substitutions can convert the uncleaved signal-anchor of sucrase-isomaltase to a cleaved signal sequence. J Biol Chem. 1992 Aug 25;267(24):16928–16933. [PubMed] [Google Scholar]
  12. Hogue B. G., Nayak D. P. Deletion mutation in the signal anchor domain activates cleavage of the influenza virus neuraminidase, a type II transmembrane protein. J Gen Virol. 1994 May;75(Pt 5):1015–1022. doi: 10.1099/0022-1317-75-5-1015. [DOI] [PubMed] [Google Scholar]
  13. Hong W. J., Doyle D. Molecular dissection of the NH2-terminal signal/anchor sequence of rat dipeptidyl peptidase IV. J Cell Biol. 1990 Aug;111(2):323–328. doi: 10.1083/jcb.111.2.323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lipp J., Dobberstein B. The membrane-spanning segment of invariant chain (I gamma) contains a potentially cleavable signal sequence. Cell. 1986 Sep 26;46(7):1103–1112. doi: 10.1016/0092-8674(86)90710-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lobigs M. Flavivirus premembrane protein cleavage and spike heterodimer secretion require the function of the viral proteinase NS3. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6218–6222. doi: 10.1073/pnas.90.13.6218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lobigs M. Proteolytic processing of a Murray Valley encephalitis virus non-structural polyprotein segment containing the viral proteinase: accumulation of a NS3-4A precursor which requires mature NS3 for efficient processing. J Gen Virol. 1992 Sep;73(Pt 9):2305–2312. doi: 10.1099/0022-1317-73-9-2305. [DOI] [PubMed] [Google Scholar]
  17. Markoff L., Chang A., Falgout B. Processing of flavivirus structural glycoproteins: stable membrane insertion of premembrane requires the envelope signal peptide. Virology. 1994 Nov 1;204(2):526–540. doi: 10.1006/viro.1994.1566. [DOI] [PubMed] [Google Scholar]
  18. Markoff L. In vitro processing of dengue virus structural proteins: cleavage of the pre-membrane protein. J Virol. 1989 Aug;63(8):3345–3352. doi: 10.1128/jvi.63.8.3345-3352.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nothwehr S. F., Hoeltzli S. D., Allen K. L., Lively M. O., Gordon J. I. Residues flanking the COOH-terminal C-region of a model eukaryotic signal peptide influence the site of its cleavage by signal peptidase and the extent of coupling of its co-translational translocation and proteolytic processing in vitro. J Biol Chem. 1990 Dec 15;265(35):21797–21803. [PubMed] [Google Scholar]
  20. Nowak T., Färber P. M., Wengler G., Wengler G. Analyses of the terminal sequences of West Nile virus structural proteins and of the in vitro translation of these proteins allow the proposal of a complete scheme of the proteolytic cleavages involved in their synthesis. Virology. 1989 Apr;169(2):365–376. doi: 10.1016/0042-6822(89)90162-1. [DOI] [PubMed] [Google Scholar]
  21. Preugschat F., Yao C. W., Strauss J. H. In vitro processing of dengue virus type 2 nonstructural proteins NS2A, NS2B, and NS3. J Virol. 1990 Sep;64(9):4364–4374. doi: 10.1128/jvi.64.9.4364-4374.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rapoport T. A. Transport of proteins across the endoplasmic reticulum membrane. Science. 1992 Nov 6;258(5084):931–936. doi: 10.1126/science.1332192. [DOI] [PubMed] [Google Scholar]
  23. Roy P., Chatellard C., Lemay G., Crine P., Boileau G. Transformation of the signal peptide/membrane anchor domain of a type II transmembrane protein into a cleavable signal peptide. J Biol Chem. 1993 Feb 5;268(4):2699–2704. [PubMed] [Google Scholar]
  24. Ruiz-Linares A., Cahour A., Després P., Girard M., Bouloy M. Processing of yellow fever virus polyprotein: role of cellular proteases in maturation of the structural proteins. J Virol. 1989 Oct;63(10):4199–4209. doi: 10.1128/jvi.63.10.4199-4209.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schmid S. R., Spiess M. Deletion of the amino-terminal domain of asialoglycoprotein receptor H1 allows cleavage of the internal signal sequence. J Biol Chem. 1988 Nov 15;263(32):16886–16891. [PubMed] [Google Scholar]
  26. Wengler G., Czaya G., Färber P. M., Hegemann J. H. In vitro synthesis of West Nile virus proteins indicates that the amino-terminal segment of the NS3 protein contains the active centre of the protease which cleaves the viral polyprotein after multiple basic amino acids. J Gen Virol. 1991 Apr;72(Pt 4):851–858. doi: 10.1099/0022-1317-72-4-851. [DOI] [PubMed] [Google Scholar]
  27. Westaway E. G. Flavivirus replication strategy. Adv Virus Res. 1987;33:45–90. doi: 10.1016/s0065-3527(08)60316-4. [DOI] [PubMed] [Google Scholar]
  28. Yamshchikov V. F., Compans R. W. Formation of the flavivirus envelope: role of the viral NS2B-NS3 protease. J Virol. 1995 Apr;69(4):1995–2003. doi: 10.1128/jvi.69.4.1995-2003.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Yamshchikov V. F., Compans R. W. Processing of the intracellular form of the west Nile virus capsid protein by the viral NS2B-NS3 protease: an in vitro study. J Virol. 1994 Sep;68(9):5765–5771. doi: 10.1128/jvi.68.9.5765-5771.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yamshchikov V. F., Compans R. W. Regulation of the late events in flavivirus protein processing and maturation. Virology. 1993 Jan;192(1):38–51. doi: 10.1006/viro.1993.1006. [DOI] [PubMed] [Google Scholar]
  31. Yang X. F., Chatellard C., Lazure C., Crine P., Boileau G. Insertion of hydrophilic amino acid residues in the signal peptide/membrane anchor domain of neprilysin (neutral endopeptidase-24.11) results in its cleavage: role of the position of insertion. Arch Biochem Biophys. 1994 Dec;315(2):382–386. doi: 10.1006/abbi.1994.1514. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES