Skip to main content
SpringerLink
Log in

Semliki Forest virus capsid protein expressed by a baculovirus recombinant

  • Original Papers
  • Published:
Archives of Virology Aims and scope Submit manuscript

Summary

We have constructed a recombinant baculovirus which expressed the Semliki Forest virus (SFV) capsid (C) gene as a fusion protein under the control of the polyhedrin gene promoter. The sequence coding for C and part of the envelope E3 region were expressed as a polyprotein precursor.Spodoptera frugiperda (Sf9) insect cells infected with the recombinant virus produced a protein reacting with polyclonal rabbit antiserum that had been raised against the 33 kDa authentic C protein purified from SFV. Primer extension analyses showed that transcription from the polyhedrin promoter started in the late/very late consensus initiation motif. Due to its autoprotease activity, the recombinant C protein was contranslationally cleaved within its C-terminus and the mature protein remained undegraded for long periods of time.Spodoptera frugiperda cells infected with recombinant virus synthesized large amounts of C protein. The recombinant protein was karyophilic as is authentic SFV C protein and it was translocated into the nucleus; there, it was associated with nucleolus-like structures.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Boege U, Wengler G, Wittmann-Liebold B (1983) The core protein of alphaviruses. 1. Purification of peptides and complete amino-acid sequence of Semliki Forest virus core protein. Eur J Biochem 133: 405–414

    Google Scholar 

  2. Brzeski H, Clegg JCS, Atkins GJ, Kennedy SIT (1978) Regulation of the synthesis of Sindbis virus-specified RNA: role of the virion core protein. J Gen Virol 38: 461–470

    Google Scholar 

  3. Carstens EB, Tjia ST, Doerfler W (1979) Infection ofSpodoptera frugiperda cells withAutographa californica nuclear polyhedrosis virus. 1. Synthesis of intracellular proteins after virus infection. Virology 99: 386–398

    Google Scholar 

  4. Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18: 5294–5299

    Google Scholar 

  5. Elgizoli M, Dai Y, Kempf C, Koblet H, Michel MR (1989) Semliki Forest virus capsid protein acts as a pleiotropic regulator of host cellular protein synthesis. J Virol 63: 2921–2928

    Google Scholar 

  6. Fuller SD (1987) The T-4 envelope of Sindbis virus is organized by interactions with a complementary T-3 capsid. Cell 48: 923–934

    Google Scholar 

  7. Garoff H, Frischauf A-M, Simons K, Lehrach H, Delius H (1980) The capsid protein of Semliki Forest virus has clusters of basic amino acids and prolines in its aminoterminal region. Proc Natl Acad Sci USA 77: 6376–6380

    Google Scholar 

  8. Garoff H, Kondor-Koch C, Riedel H (1982) Structure and assembly of alphaviruses. Curr Top Microbiol Immunol 99: 1–50

    Google Scholar 

  9. Gearing KL, Possee RD (1990) Functional analysis of a 603 nucleotide open reading frame upstream of the polyhedrin gene ofAutographa californica nuclear polyhedrosis virus. J Gen Virol 71: 251–262

    Google Scholar 

  10. Howard SC, Ayres MD, Possee RD (1986) Mapping the 5′ and 3′ ends ofAutographa californica nuclear polyhedrosis virus polyhedrin mRNA. Virus Res 5: 109–119

    Google Scholar 

  11. Iddekinge BJL Hooft van, Smith GE, Summers MD (1983) Nucleotide sequence of the polyhedrin gene ofAutographa californica nuclear polyhedrosis virus. Virology 131: 561–565

    Google Scholar 

  12. Jalanko A (1985) Expression of Semliki Forest virus capsid protein from SV 40 recombinant virus. FEBS Lett 186: 59–64

    Google Scholar 

  13. Knudson DL, Harrap KA (1976) Replication of a nuclear polyhedrosis virus in a continuous cell culture ofSpodoptera frugiperda: microscopy study of the sequence of events of the virus infection. J Virol 17: 254–268

    Google Scholar 

  14. Koblet H (1990) The “merry-go-round”: alphaviruses between vertebrate and invertebrate cells. Adv Virus Res 38: 343–402

    Google Scholar 

  15. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

    Google Scholar 

  16. Liljeström P, Lusa S, Huylebroeck D, Garoff H (1991) In vitro mutagenesis of a full-length cDNA clone of Semliki Forest virus: the small 6,000-molecular-weight membrane protein modulates virus release. J Virol 65: 4107–4113

    Google Scholar 

  17. Luckow VA, Summers MD (1988) Trends in the development of baculovirus expression vectors. BioTechnology 6: 47–55

    Google Scholar 

  18. Melancon P, Garoff H (1987) Processing of the Semliki Forest virus structural poly-protein: role of the capsid protease. J Virol 61: 1301–1309

    Google Scholar 

  19. Michel MR, Elgizoli M, Dai Y, Jakob R, Koblet H, Arrigo A-P (1990) Karyophilic properties of Semliki Forest virus nucleocapsid protein. J Virol 64: 5123–5131

    Google Scholar 

  20. Oker-Blom C, Summers MD (1989) Expression of Sindbis virus 26 S cDNA inSpodoptera frugiperda (Sf9) cells, using a baculovirus expression vector. J Virol 63: 1256–1264

    Google Scholar 

  21. Possee RD, Howard SC (1987) Analysis of the polyhedrin gene promoter of theAutographa californica nuclear polyhedrosis virus. Nucleic Acids Res 15: 10233–10248

    Google Scholar 

  22. Possee RD, Sun T-P, Howard SC, Ayres MD, Hill-Perkins M, Gearing KL (1991) Nucleotide sequence of theAutographa californica nuclear polyhedrosis 9.4kbpEco RI-I and -R (polyhedrin gene) region. Virology 185: 229–241

    Google Scholar 

  23. Sambrook J, Fritsch EF, Maniatis T (1990) Molecular cloning: A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor

    Google Scholar 

  24. Simons K, Warren G (1984) Semliki Forest virus: a probe for membrane traffic in the animal cell. Adv Protein Chem 36: 79–132

    Google Scholar 

  25. Robin Y (1981 a) Semliki Forest viral fever. In: Steele JH (ed) Viral zoonoses. CRC Press, Boca Raton, pp 239–241 (CRB Handbook, Ser. Zoonoses, vol 1).

    Google Scholar 

  26. Stoflet ES, Koeberl DD, Sarkar G, Sommer SS (1988) Genomic amplification with transcript sequencing. Science 239: 491–494

    Google Scholar 

  27. Summers MD, Smith GE (1987) A manual of methods for baculovirus vectors and insect cell culture procedures. Tex Agric Exper Stat Bull no. 1555

  28. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354

    Google Scholar 

  29. Ulmanen I, Söderlund H, Kääriäinen (1976) Semliki Forest virus capsid protein associates with the 60 S ribosomal subunit in infected cells. J Virol 20: 203–210

    Google Scholar 

  30. Wengler G (1987) The mode of assembly of alphavirus cores implies a mechanism for the disassembly of the cores in the early stages of infection. Arch Virol 94: 1–14

    Google Scholar 

Download references

Author information

Author notes

  1. T. Nishimura

    Present address: Department of Anatomy, Aichi Medical University, Nagakute-cho, Japan

Authors and Affiliations

  1. Institute of Medical Microbiology, University of Berne, Berne, Switzerland

    D. Favre, E. Studer, T. Nishimura & M. R. Michel

  2. Institute for Clinical Microbiology and Immunology, St-Gallen, Switzerland

    M. Weitz

Authors
  1. D. Favre
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. Studer
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. T. Nishimura
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. Weitz
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. R. Michel
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Favre, D., Studer, E., Nishimura, T. et al. Semliki Forest virus capsid protein expressed by a baculovirus recombinant. Archives of Virology 132, 307–319 (1993). https://doi.org/10.1007/BF01309541

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01309541

Keywords

Search

Navigation