1887

Abstract

SUMMARY

Monoclonal antibodies were developed that are specific for Rous sarcoma virus structural, polymerase (reverse transcriptase) and transforming proteins. The monoclonal antibodies were shown to bind to purified virus proteins in an indirect I-labelled Protein A binding assay suitable for screening even very large numbers of hybridomas. Additional tests for specificity included radioimmunoprecipitation of purified virus structural proteins P12 and P27, of reverse transcriptase subunits and , and of the transforming protein pp60. Pilot immunofluorescence and protein kinase assays of the expression of virus proteins in avian and mammalian cells infected by wild-type virus as well as by temperature-sensitive, transformation-defective virus mutants revealed that synthesis of virus structural and transforming proteins is hardly affected by changes in temperature, whereas the pp60-associated kinase activity is temperature-sensitive in cells infected by most, but not all the virus mutants.

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1985-04-01
2024-04-25
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References

  1. Balduzzi P., Beamand J. A., Christensen J. R., Pearson Y. M., Wyke J. A. 1978; Provisional mapping of transformation defective temperature sensitive mutants of Rous sarcoma virus. In Avian RNA Tumor Viruses pp 112–121 Edited by Barlati S., de Giuli-Morghen C. Padua: Piccin Medical Books;
    [Google Scholar]
  2. Bishop J. M., Varmus H. 1982; Functions and origins of retroviral transforming genes. In RNA Tumor Viruses pp 999–1108 Edited by Weiss R., Teich N., Varmus H., Coffin J. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  3. Boettiger D., Love D. N., Weiss R. A. 1975; Virus envelope markers in mammalian tropism of avian RNA tumor viruses. Journal of Virology 15:108–114
    [Google Scholar]
  4. Brugge J. S., Erikson R. L. 1977; Identification of a transformation-specific antigen induced by avian sarcoma virus. Nature, London 269:346–348
    [Google Scholar]
  5. Bunte T., Owada M. K., Donner P., Boschek C. B., Moelling K. 1981; Association of the transformation-specific protein pp60src with the membrane of an avian sarcoma virus. Journal of Virology 33:1034–1047
    [Google Scholar]
  6. Chiswell D. J., Enrietto P. J., Evans S., Quade K., Wyke J. A. 1982; Molecular mechanisms involved in morphological variation of avian sarcoma virus-infected rat cells. Virology 116:428–440
    [Google Scholar]
  7. Collett M. S., Erikson R. L. 1978; Protein kinase activity associated with the avian sarcoma virus src gene product. Proceedings of the National Academy of Sciences, U,. S,. A 75:2021–2024
    [Google Scholar]
  8. Donner P., Bunte T., Owada M. K., Moelling K. 1981; Biochemical characterization of pp60src-associated protein kinase from avian sarcoma virus Schmidt-Ruppin strain. Journal of Biological Chemistry 256:8786–8794
    [Google Scholar]
  9. Ellwart-Tschürtz M., Kurth R. 1981; Demonstration of avian sarcoma virus-coded pp60src in vivo and the anti-pp60src immune response in chickens. Journal of Virology 39:950–953
    [Google Scholar]
  10. Fazekas De St Groth S., Scheidegger D. 1980; Production of monoclonal antibodies: strategy and tactics. Journal of Immunological Methods 35:1–21
    [Google Scholar]
  11. Fincham V. J., Chiswell D. J., Wyke J. A. 1982; Mapping of nonconditional and conditional mutants in the src-gene of Prague strain Rous sarcoma virus. Virology 116:72–83
    [Google Scholar]
  12. Galfré G., Milstein C. 1981; Preparation of monoclonal antibodies. Methods in Enzymology 73:3–46
    [Google Scholar]
  13. Huesgen A., Huesgen G., Stegmann L., Kurth R. 1980; The immune response against the ASV-coded src-gene product in syngeneic mice. Journal of General Virology 48:401–406
    [Google Scholar]
  14. Hunter T., Sefton B. M. 1980; Transforming gene product of Rous sarcoma virus phosphorylates tyrosine. Proceedings of the National Academy of Sciences, U,. S,. A 77:1311–1315
    [Google Scholar]
  15. Köhler G. 1981; The technique of hybridoma production. In Immunological Methods vol 2: pp 285–298 Edited by Lefkovits I., Pernis B. New York: Academic Press;
    [Google Scholar]
  16. Kurth R. 1983; Oncogenes in retroviruses and cells. Naturwissenschaften 70:439–450
    [Google Scholar]
  17. Kurth R., Mikschy U. 1978; Human antibodies reactive with purified envelope antigens of primate type C tumor viruses. Proceedings of the National Academy of Sciences U,. S,. A 75:5692–5696
    [Google Scholar]
  18. Lipsich L. A., Lewis A. J., Brugge J. S. 1983; Isolation of monoclonal antibodies that recognize the transforming proteins of avian sarcoma viruses. Journal of Virology 48:352–360
    [Google Scholar]
  19. Martin G. S. 1970; Rous sarcoma virus: a function required for the maintenance of the transformed state. Nature. London 227:1021–1023
    [Google Scholar]
  20. Moelling K., Sykora K. W., Dittmar K. E., Scott A., Watson K. F. 1979; The isolation of avian viral RNA and polypeptides. Journal of Biological Chemistry 254:3738–3742
    [Google Scholar]
  21. Nigg E. A., Walter G., Singer S. J. 1982; On the nature of crossreactions observed with antibodies directed to defined epitopes. Proceedings of the National Academy of Sciences, U.S.A 79:5939–5943
    [Google Scholar]
  22. Parsons S. J., Riley S. C., Mullen E. E., Brock E. J., Benjamin D. C., Kuehl W. M., Parsons J. T. 1979; Immune response to the src-gene product in mice bearing tumors induced by injection of avian sarcoma virus-transformed mouse cells. Journal of Virology 32:40–46
    [Google Scholar]
  23. Parsons S. J., McCarley D. J., Ely C. M., Benjamin D. C., Parsons J. T. 1983; Isolation and partial characterization of a monoclonal antibody to the Rous sarcoma virus transforming protein pp60src. Journal of Virology 45:1190–1194
    [Google Scholar]
  24. Richert N. D. 1982; Monoclonal antibody specific for avian sarcoma virus structural protein p27. Journal of General Virology 62:385–392
    [Google Scholar]
  25. Schäfer W. 1979; Der Mause-Inzuchtstamm STU. Entwicklung und Eigenschaften. Zeitschrijt für Naturforschung 34c:306–309
    [Google Scholar]
  26. Sovová V., Cerná H., Dostálová V., Hlozánek I. 1980; Use of the hybridoma technique for antibody production to group-specific antigens of avian oncornaviruses. Folia biologica, Praha 26:366–368
    [Google Scholar]
  27. Tanaka T., Kurth R. 1984a; Monoclonal antibodies specific for the avian sarcoma virus transforming protein pp60src. Virology 133:202–210
    [Google Scholar]
  28. Tanaka T., Kurth R. 1984b; A small scale indirect 125iodine-labelled protein A binding assay for detection of monoclonal antibodies against avian oncoviral proteins. Journal of Immunological Methods 75:217–225
    [Google Scholar]
  29. Varmus H. E., Guntaka R. V., Deng C. T., Bishop J. M. 1975; Synthesis, structure and function of avian sarcoma virus-specific DNA in permissive and nonpermissive cells. Cold Spring Harbor Symposia on Quantitative Biology 39:987–996
    [Google Scholar]
  30. Weis J., Smith D., Burns N., Furcht L., Faras A. 1983; Production and characterization of monoclonal antibodies against avian retrovirus reverse transcriptase. Journal of Virology 45:859–863
    [Google Scholar]
  31. Wyke J. A. 1973a; The selective isolation of temperature-sensitive mutants of Rous sarcoma virus. Virology 52:587–590
    [Google Scholar]
  32. Wyke J. A. 1973b; Complementation of transforming functions by temperature-sensitive mutants of avian sarcoma virus. Virology 54:28–36
    [Google Scholar]
  33. Wyke J. A., Kurth R. 1978; Reversion of temperature sensitive transformation mutants of Rous sarcoma virus and its effect on the expression of tumour specific surface antigen. Journal of General Virology 40:701–704
    [Google Scholar]
  34. Wyke J. A., Quade K. 1980; Infection of rat cells by avian sarcoma virus: factors affecting transformation and subsequent reversion. Virology 106:217–233
    [Google Scholar]
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