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Cell-free translation ofDrosophila C virus RNA: Identification of a virus protease activity involved in capsid protein synthesis and further studies onin vitro processing of Cricket paralysis virus specified proteins

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Summary

Drosophila C virus RNA acted as mRNA in rabbit reticulocyte lysates and directed the synthesis of at least one capsid protein and a number of higher molecular weight proteins. Kinetic analysis by pulse-chase experiments showed that a number of high molecular weight products acted as precursors to the capsid protein(s). Various dilution experiments were performed which showed that the virus specified a protease activity essential for the correct processing of precursors to give the capsid protein(s). A similar result was obtained with Cricket paralysis virus, and mixing experiments showed that the protease activity specified by one virus could perform some of the cleavages resulting in the production of the capsid proteins of the other virus. Some of the cleavages involving the highest molecular weight precursors could not be performed by the protease activity of the other virus. We could find no evidence for intramolecular cleavage of the capsids precursors of either of the viruses.

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References

  1. Cleveland, D. W., Fischer, S. G., Kirschner, M. W., Laemmli, U. K.: Peptide mapping by limited proteolysis in sodium dodecyl sulphate and analysis by gel electrophoresis. J. Biol. Chem.252, 1102–1106 (1977).

    Google Scholar 

  2. Kitamura, N., Semler, B. L., Rothberg, P. G., Larsen, G. R., Adler, C. J., Dorner, A. J., Emini, E. A., Hanecak, R., Lee, J. J., van der Werf, S., Anderson, C. W., Wimmer, E.: Primary sturcture gene organization and polypeptide, expression of poliovirus RNA. Nature (London)291, 547–553 (1981).

    Google Scholar 

  3. Korant, B. D.: Cleavage of viral precursor proteinsin vivo andin vitro. J. Virol.10, 751–759 (1972).

    Google Scholar 

  4. Laemmli, U. K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London)227, 680–685 (1970).

    Google Scholar 

  5. Larsen, G. R., Anderson, C. W., Dorner, A. J., Semler, B. L., Wimmer, E.: Cleavage sites within the poliovirus capsid protein precursors. J. Virol.41, 340–344 (1982).

    Google Scholar 

  6. Laskey, R. Mills, A.: Quantitative film detection of3H and14C in polyacrylamide gels by fluorography. Eur. J. Biochem.56, 335–341 (1975).

    Google Scholar 

  7. Matthews, R. E. F.: Fowel report of the International Committee on Taxonomy of Viruses. Intervirol.17, 1–200 (1982).

    Google Scholar 

  8. Moore, N. F., Kearns, A., Pullin, J. S. K.: Characterization of Cricket paralysis. virus-induced polypeptides inDrosophila cells. J. Virol.33, 1–9 (1980).

    Google Scholar 

  9. Moore, N. F., Pullin, J. S. K.: Plaque purification of Cricket paralysis virus using an agar overlay onDrosophila cells. J. Invertebr. Pathol.39, 10–14 (1982).

    Google Scholar 

  10. Moore, N. F., Pullin, J. S. K., Reavy, B.: The intracellular proteins induced by Cricket paralysis virus inDrosophila cells: the effect of protease inhibitors and amino acid analogues. Arch. Virol.70, 1–9 (1981).

    Google Scholar 

  11. Moore, N. F., Reavy, B., Pullin, J. S. K.: Processing of Cricket paralysis virus induced polypeptides inDrosophila cells: production of high molecular weight polypeptides by treatment with iodoacetamide. Arch. Virol.68, 1–8 (1981).

    Google Scholar 

  12. Moore, N. F., Reavy, B., Pullin, J. S. K., Plus, N.: The polypeptides induced inDrosophila cells byDrosophila C virus (strain Ouarzazate). Virology112, 411–416 (1981).

    Google Scholar 

  13. Palmenberg, A. C., Rueckert, R. R.: Evidence for intramolecular self-cleavage of picornaviral replicase precursors. J. Virol.41, 244–249 (1982).

    Google Scholar 

  14. Pelham, H. R. B.: Synthesis and proteolytic processing of cowpea mosaic virus proteins in reticulocyte lysates. Virology96, 463–477 (1979).

    Google Scholar 

  15. Pelham, H. R. B., Jackson, R. J.: An efficient mRNA-dependent translation system from reticulocyte lysates. Eur. J. Biochem.67, 247–256 (1976).

    Google Scholar 

  16. Plus, N., Croizier, G., Reinganum, C., Scotti, P. D.: Cricket paralysis virus andDrosophila C virus: serological analysis and comparison of capsid polypeptides and host range. J. Invertebr. Pathol.31, 296–302 (1978).

    Google Scholar 

  17. Reavy, B., Moore, N. F.:In vitro translation of Cricket paralysis virus RNA. Arch. Virol.67, 175–180 (1981).

    Google Scholar 

  18. Reavy, B., Moore, N. F.: Cell-free translation of Cricket paralysis virus RNA: analysis of the synthesis and processing of virus-specified proteins. J. gen. Virol.55, 429–438 (1981).

    Google Scholar 

  19. Reavy, B., Moore, N. F.: The replication of small RNA-containing viruses of insects. Microbiologica5, 63–84 (1982).

    Google Scholar 

  20. Rueckert, R. R.: On the structure and morphogenesis of picornaviruses. In:Fraenkel-Conrat, H., Wagner, R. R. (eds.), Comprehensive Virology, Vol. 6, 131–213. New York: Plenum Publishing Corp. 1976.

    Google Scholar 

  21. Scotti, P. D.: Cricket paralysis virus replicates in culturedDrosophila cells. Intervirol.6, 333–342 (1976).

    Google Scholar 

  22. Scotti, P. D.: End-point dilution and plaque-assay methods for titration of Cricket paralysis virus in culturedDrosophila cells. J. gen. Virol.35, 393–396 (1977).

    Google Scholar 

  23. Semler, B. L., Hanecak, R., Anderson, C. W., Wimmer, E.: Cleavage sites in the polypeptide precursors of poliovirus protein P2-X. Virology114, 589–594 (1981).

    Google Scholar 

  24. Shih, D. S., Shih, C. T., Kew, O., Pallansch, M., Rueckert, R. R., Kaesberg, P.: Cell-free synthesis and processing of the proteins of poliovirus. Proc. Nat. Acad. Sci. U.S.A.75, 5807–5811 (1978).

    Google Scholar 

  25. Svitkin, Y. V., Gorbalenya, A. E., Kazachkov, Y. A., Agol, V. I.: Encephalomyocarditis virus-specific polypeptide p22 possessing a proteolytic activity. Preliminary mapping on the viral genome. FEBS Letts.108, 6–9 (1979).

    Google Scholar 

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Reavy, B., Moore, N.F. Cell-free translation ofDrosophila C virus RNA: Identification of a virus protease activity involved in capsid protein synthesis and further studies onin vitro processing of Cricket paralysis virus specified proteins. Archives of Virology 76, 101–115 (1983). https://doi.org/10.1007/BF01311694

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