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In vitro reconstitution of rotavirus transcriptional activity using viral cores and recombinant baculovirus expressed VP 6

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Summary

Purified baculovirus-expressed group A rotavirus VP 6 polypeptide was shown to be active in the recovery of the transcriptase activity associated with the reconstitution of the single-shelled rotavirus particle. Recombinant VP 6 polypeptide was able to restore the transcriptional activity in purified viral cores from both SA-11 and RF rotavirus strains. Recombinant group C VP 6 (Cowden strain) is capable of binding as a trimer to group A viral core particles but unable to restore the transcriptase activity, suggesting that the binding of the polypeptide to cores is not the only requirement to restore the transcriptase activity. The VP 6 group A polypeptide was shown to bind as a monomer to viral cores, indicating that trimerization of VP 6 may be not required for reconstitution of the polymerase activity.

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References

  1. Cukor G, Blacklow NR (1984) Human viral gastroenteritis. Microbiol Rev 48: 157–179

    Google Scholar 

  2. Estes MK, Palmer EL, Obijeski JF (1983) Rotaviruses: a review. Curr Top Microbiol Immunol 105: 123–184

    Google Scholar 

  3. Estes MK, Cohen J (1989) Rotavirus gene structure and function. Microbiol Rev 53: 410–449

    Google Scholar 

  4. Bican P, Cohen J, Charpilienne A, Scherrer R (1982) Purification and characterization of bovine rotavirus cores. J Virol 43: 1113–1117

    Google Scholar 

  5. Valenzuela S, Pizarro JM, Sandino AM, Vasquez M, Fernandez J, Hernandez O, Patton J, Spencer E (1991) Photoaffinity labeling of rotavirus VP 1 with 8-azido-ATP: identification of the viral RNA polymerase. J Virol 65: 3964–3967

    Google Scholar 

  6. Mansell E, Patton J (1990) Rotavirus RNA replication: VP 2 but not VP 6, is necessary for viral replicase activity. J Virol 64: 4988–4996

    Google Scholar 

  7. Patton J, Gallegos C (1988) Structure and protein composition of the rotavirus replicase particle. Virology 166: 358–365

    Google Scholar 

  8. Pizarro JL, Sandino AM, Pizarro JM, Fernandez J, Spencer E (1991) Characterization of rotavirus guanylyltransferase activity associated to VP 3. J Gen Virol 72: 325–332

    Google Scholar 

  9. Pizarro JM, Pizarro JL, Fernandez J, Sandino AM, Spencer E (1991) Effect of nucleotide analogs on rotavirus transcription and replication. Virology 184: 768–772

    Google Scholar 

  10. Novo E, Esparza J (1981) Composition and topography of structural polypeptides of bovine rotavirus. J Gen Virol 56: 325–335

    Google Scholar 

  11. Sabara M, Ready KFM, Frenchick PJ, Babiuk LA (1987) Biochemical evidence for the oligomeric arrangement of bovine rotaviruses nucleocapsid protein and its possible significance in the immunogenicity of this protein. J Gen Virol 68: 123–133

    Google Scholar 

  12. Sabara M, Parker M, Aha P, Cosco C, Gibbons E, Parsons S, Babiuk LA (1991) Assembly of double-shelled rotaviruslike particles by simultaneous expression of recombinant VP 6-protein and VP 7-protein. J Virol 65: 6994–6997

    Google Scholar 

  13. Sandino AM, Jashés M, Faúndez G, Spencer E (1986) Role of the inner protein capsid on “in vitro” human rotavirus transcription. J Virol 60: 797–802

    Google Scholar 

  14. Sandino AM, Pizarro J, Fernandez J, Fellay C, Spencer E (1988) Involvement of structural and nonstructural polypeptides in rotavirus RNA synthesis. Arch Biol Med Exper 21: 381–392

    Google Scholar 

  15. Estes M, Crawford M, Penaranda B, Petrie J, Chan W, Ericson B, Smith G, Summers D (1987) Synthesis and immunogenity of the rotavirus major capsid antigen using a baculovirus expression system. J Virol 61: 1488–1494

    Google Scholar 

  16. Tosser G, Labbe M, Bremont M, Cohen J (1992) Expression of the major capsid protein VP 6 of group C rotavirus and synthesis of chimeric single-shelled particles using recombinant baculoviruses. J Virol 66: 5825–5831

    Google Scholar 

  17. Labbe M, Charpiliene A, Crawford S, Estes MK, Cohen J (1991) Expression of rotavirus VP 2 produces empty corelike particles. J Virol 65: 2946–2952

    Google Scholar 

  18. Clapp L, Patton J (1991) Rotavirus morphogenesis: domains in the mayor inner capsid protein essential for binding to single-shelled particles and for trimerization. Virology 180: 697–708

    Google Scholar 

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Authors and Affiliations

  1. Laboratoire de Virologie, Facultés de Médicine et Pharmacie, Université de Bourgogne, Dijon

    Evelyne Kohli, P. Pothier & E. Spencer

  2. Unité de Virologie et Immunologie Moléculaire, C.R.J., INRA Jouy-en-Josas, France

    Gwenola Tosser & Jean Cohen

  3. Unidad de Virología, INTA Universidad de Chile, Santiago, Chile

    Ana Maria Sandino

Authors
  1. Evelyne Kohli
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  2. P. Pothier
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  3. Gwenola Tosser
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  4. Jean Cohen
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  5. Ana Maria Sandino
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  6. E. Spencer
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Kohli, E., Pothier, P., Tosser, G. et al. In vitro reconstitution of rotavirus transcriptional activity using viral cores and recombinant baculovirus expressed VP 6. Archives of Virology 133, 451–458 (1993). https://doi.org/10.1007/BF01313782

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  • DOI: https://doi.org/10.1007/BF01313782

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