1887

Abstract

Summary

Double-stranded RNA (dsRNA) from virus particles isolated from a P4 killer strain (isolate 77) of , which secretes a protein toxin of apparent approx. 10000 (10K) was resolved by PAGE into eight segments with sizes (kbp) 6·7 (H1), 4·5 (H2), 3·2 (H3a), 3·1 (H3b), 2·6 (H4), 0·98 (M2-4), 0·92 (M3-4) and 0·36 (L1). A subculture, designated 77-3A, was found to have lost the H2 and M3-4 segments, but still produced virus particles and protein toxin. Northern hybridization showed that dsRNA segments M2-4 and L1 are related but no homology was detected between H1, H3a, H3b and H4, or between any of these H segments and M2-4 or L1. When individual denatured dsRNA segments were translated in a reticulocyte lysate system H1, H3a, H3b and H4 each gave rise to polypeptides in the range 100K to 128K. Polypeptides of apparent 108K and 128K, encoded by H3b, and 100K, encoded by H4, were immunoprecipitated by antiserum to purified virus particles from subculture 77-3A. translation of denatured M2-4 gave rise to polypeptides of apparent 26K and 13K which were immunoprecipitated by antiserum to the secreted protein toxin. No translation product was detected from denatured L1.

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1988-05-01
2024-05-03
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References

  1. BEN-ZVI B. S., KOLTIN Y., MEVARECH M., TAMARKIN A. 1984; RNA polymerase activity in virions from Ustilago maydis. Molecular and Cellular Biology 4:188–194
    [Google Scholar]
  2. BOZARTH R. F., HARLEY E. H. 1976; The electrophoretic mobility of dsRNA in Polyacrylamide gels as a function of molecular weight. Biochimica et biophysica acta 432:329–335
    [Google Scholar]
  3. BOZARTH R. F., WOOD H. A., MANDELBROT A. 1971; The Penicillium stoloniferum virus-like complex: two similar double-stranded RNA virus-like particles in a single cell. Virology 45:516–523
    [Google Scholar]
  4. BOZARTH R. F., KOLTIN Y., WEISSMAN M. B., PARKER R. L., DALTON R. E., STEINLAUF R. 1981; The molecular weight and packaging of dsRNA in the mycovirus from Ustilago maydis killer strains. Virology 113:492–502
    [Google Scholar]
  5. BRUENN I. 1986 The killer systems of Saccharomyces cerevisiae and other yeasts. Fungal Virology85–108 Edited by Buck K. W. Boca Raton: CRC Press;
    [Google Scholar]
  6. BUCK K. W. 1986 Fungal virology - an overview. Fungal Virology1–84 Edited by Buck K. W. Boca Raton: CRC Press;
    [Google Scholar]
  7. BUCK K. W., GIRVAN R. F. 1977; Comparison of the biophysical properties of Penicillium cyaneo-fulvum virus and Penicillium chrysogenum virus. Journal of General Virology 34:145–154
    [Google Scholar]
  8. BUCK K. W., RATH G. 1975; Biophysical and biochemical properties of two viruses isolated from Aspergillus foetidus. Journal of General Virology 27:211–224
    [Google Scholar]
  9. BUCK K. W., RATTI G. 1977; Molecular weight of double-stranded RNA: a re-examination of Aspergillus foetidus virus S RNA components. Journal of General Virology 37:215–219
    [Google Scholar]
  10. BUCK K. W., ALMOND M. R., MCFADDEN J. J. P., ROMANOS M. A., RAWLINSON C.J. 1981; Properties of thirteen viruses and virus variants obtained from eight isolates of the wheat take-all fungus Gaeumannomyces graminis var. tritici. Journal of General Virology 53:235–245
    [Google Scholar]
  11. CHAMBERLAIN J. P. 1979; Fluorographic detection of radioactivity in Polyacrylamide gels with the water-soluble fluor, sodium salicylate. Analytical Biochemistry 98:132–135
    [Google Scholar]
  12. FIELD L. J., BRUENN J. A., CHANG T. H., PINHASI O., KOLTIN Y. 1983; Two Ustilago maydis viral dsRNAs of different size code for the same product. Nucleic Acids Research 11:2765–2778
    [Google Scholar]
  13. FRANKLIN R. M. 1966; Purification and properties of the replicative intermediate of the RNA bacteriophage R17. Proceedings of the National Academy of Sciences, U.S.A 55:1504–1511
    [Google Scholar]
  14. JORDAN R. L., DODDS J. A. 1983; Hybridization of 5′ end labelled RNA to plant viral RNA in agarose and acrylamide gels. Plant Molecular Biology Reporter 1:31–37
    [Google Scholar]
  15. KOLTIN Y. 1986 The killer systems of Ustilago maydis. Fungal Virology109–141 Edited by Buck K. W. Boca Raton: CRC Press;
    [Google Scholar]
  16. KOLTIN Y., DAY P. R. 1976; Inheritance of killer phenotypes and double-stranded RNA in Ustilago maydis. Proceedings of the National Academy of Sciences, U.S.A 73:594–598
    [Google Scholar]
  17. KOLTIN Y., KANDEL J. S. 1978; Killer phenomenon in Ustilago maydis: the organisation of the viral genome. Genetics 88:267–276
    [Google Scholar]
  18. KOLTIN Y., MAYER I., STEINLAUF R. 1978; Killer phenomenon in Ustilago maydis: mapping viral functions. Molecular and General Genetics 166:181–186
    [Google Scholar]
  19. KOLTIN Y., LEVINE R., PEERY T. 1980; Assignment of functions to segments of the dsRNA genome of the Ustilago virus. Molecular and General Genetics 178:173–178
    [Google Scholar]
  20. LAEMMLI U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  21. LAMPSON G. P., TYTELL A. A., FIELD A. K., NEMES M. M., HILLEMAN M. R. 1967; Inducers of interferon and host resistance. I. Double-stranded RNA from extracts of Penicillium funiculosum. Proceedings of the National Academy of Sciences, U.S.A 58:782–786
    [Google Scholar]
  22. LEVINE R., KOLTIN Y., KANDEL J. 1979; Nuclease activity associated with the Ustilago maydis virus induced killer proteins. Nucleic Acids Research 6:3717–3731
    [Google Scholar]
  23. MANIATIS T., FRITSCH E. F., SAMBROOK J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  24. PEACOCK A. C., DINGMAN C W. 1968; Molecular weight estimation and separation of ribonucleic acid by electrophoresis in agarose-acrylamide composite gels. Biochemistry 7:668–674
    [Google Scholar]
  25. PEERY T., KOLTIN Y., TAMARKIN A. 1982; Mapping the immunity function of the Ustilago maydis P1 virus. Plasmid 7:52–58
    [Google Scholar]
  26. PEERY T., SHABAT-BRAND T., STEINLAUF R., KOLTIN Y., BRUENN J. 1987; Virus-encoded toxin of Ustilago maydis: two polypeptides are essential for activity. Molecular and Cellular Biology 7:470–477
    [Google Scholar]
  27. PELHAM H. R. B., JACKSON R. J. 1976; An efficient mRNA-dependent translation system from reticulocyte lysates. European Journal of Biochemistry 67:247–256
    [Google Scholar]
  28. PODILA G. K., FLURKEY W. H., BOZARTH R. F. 1987a; Identification and comparison of viral genes coding for capsid proteins of Ustilago maydis virus. Journal of General Virology 68:2741–2750
    [Google Scholar]
  29. PODILA G. K., BOZARTH R. F., FLURKEY W. H. 1987a; Synthesis and processing of killer toxin from Ustilago maydis virus P4. Biochemical and Biophysical Research Communications 149:391–397
    [Google Scholar]
  30. RATTI G., BUCK K. W. 1972; Virus particles in Aspergillus Foetidus: a multicomponent system. Journal of General Virology 14:165–175
    [Google Scholar]
  31. SHAPIRO A. L., VINUELA E., MAIZEL J. B. 1967; Molecular weight estimations of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochemical and Biophysical Research Communications 28:815–820
    [Google Scholar]
  32. SIMON S., GORECKI M., KOLTIN Y. 1981; In vitro translation of the Ustilago virus dsRNA. Genetics 97:s76 Abstract
    [Google Scholar]
  33. STEVENS R. B. 1974 Mycological Guidebook Seattle: University of Washington Press;
    [Google Scholar]
  34. STREET J. E., CROXSON M. C, CHADDERTON W. F., BELLAMY A. R. 1982; Sequence diversity of human rotavirus strains investigated by Northern blot hybridization analysis. Journal of Virology 43:369–378
    [Google Scholar]
  35. VODKIN M., KATTERMAN F., FINK G. R. 1974; Yeast killer mutants with altered double-stranded ribonucleic acid. Journal of Bacteriology 117:681–686
    [Google Scholar]
  36. WOOD H. A., BOZARTH R. F. 1973; Heterokaryon transfer of virus-like particles and a cytoplasmically inherited determinant in Ustilago maydis. Phytopathology 63:1019–1020
    [Google Scholar]
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