1887

Abstract

Summary

Cloned DNA representing the sequence coding for the non-structural protein 3A from cucumber mosaic virus (CMV) was inserted in an expression vector containing a truncated portion of the Protein A gene from . Expressed fusion protein was purified from cell extracts by affinity chromatography on rabbit gamma globulin-conjugated Sepharose and used as an immunogen for the production of monoclonal antibodies (MAbs). Five MAbs that reacted with pXCM3A13 fusion protein in solid phase ELISA were able to immunoprecipitate specifically protein 3A from mixtures of translation products of CMV RNA. None of these antibodies reacted with the analogous protein in translation products of brome mosaic virus RNA, but all of them reacted with the 3A protein from tomato aspermy virus. Immunogold labelling of ultrathin sections of CMV-infected tobacco tissue with MAb 3H12 demonstrated that the 3A protein accumulated within the nucleoli of these cells. This location differs from that of the analogous 3A protein of alfalfa mosaic virus which is associated with the middle lamella of the cell walls of infected tobacco cells.

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1988-09-01
2022-01-20
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References

  1. AHLQUIST P., STRAUSS E. G., RICE C. M., STRAUSS J. H., HASELOFF J., ZIMMERN D. 1985; Sindbis virus proteins nsP1 and nsP2 contain homology to non-structural proteins from several RNA plant viruses. Journal of Virology 53:536–542
    [Google Scholar]
  2. ATABEKOV J. G., DOROKHOV YU. L. 1984; Plant virus-specific transport function and resistance of plants to viruses. Advances in Virus Research 29:313–364
    [Google Scholar]
  3. D’ALESSIO I. M., NOON M. C, LEY H. L. III, GERARD G. F. 1987 One-tube double-stranded cDNA synthesis using cloned m-MLV reverse transcriptase. Focus 911–4 Gaithersburg: Bethesda Research Laboratories;
    [Google Scholar]
  4. FRANCKL R. I. B., MOSSOP D. W., HATTA T. 1979; Cucumber mosaic virus. CMI/AAB Descriptions of Plant Viruses 213:
    [Google Scholar]
  5. GODEFROY-COLBURN T., GAGEY M.-J., BERNA A., STUSSI-GARAUD C. 1986; A non-structural protein of alfalfa mosaic virus in the walls of infected tobacco cells. Journal of General Virology 67:2233–2239
    [Google Scholar]
  6. GORDON K. H. J., GILL D. S., SYMONS R. H. 1982; Highly purified cucumber mosaic virus-induced RNA-dependent RNA polymerase does not contain any of the full length translation products of the genomic RNAs. Virology 123:284–295
    [Google Scholar]
  7. GOULD A. R., SYMONS R. H. 1982; Cucumber mosaic virus RNA 3: determination of the nucleotide sequence provides the amino acid sequences of protein 3A and viral coat protein. European Journal of Biochemistry 126:217–226
    [Google Scholar]
  8. KLBERSTIS P. A., LOESCH-FRIES L. S., HALL T. C. 1981; Viral protein synthesis in barley protoplasts inoculated with native and fractionated brome mosaic virus RNA. Virology 112:804–808
    [Google Scholar]
  9. LAEMMLI U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  10. LEONARD D. A., ZAITLIN M. 1982; A temperature-sensitive strain of tobacco mosaic virus defective in cell-to-cell movement generates an altered viral-coded protein. Virology 117:416–424
    [Google Scholar]
  11. MANIATIS T., FRITSCH E. F., SAMBROOK J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  12. MESHI T., OHNO T., OKADA Y. 1982; Nucleotide sequence of the 30K protein cistron of cowpea strain of tobacco mosaic virus. Nucleic Acids Research 10:6111–6117
    [Google Scholar]
  13. MURTHY M. R. N. 1983; Comparison of the nucleotide sequences of cucumber mosaic virus and brome mosaic virus. Journal of Molecular Biology 168:469–475
    [Google Scholar]
  14. NASSUTH A., ALBLAS F., BOL J. F. 1981; Localization of genetic information involved in the replication of alfalfa mosaic virus. Journal of General Virology 53:207–214
    [Google Scholar]
  15. OHNO T., TAKAMATSU N., MESHI T., OKADA Y., NISHIGUCHI M., KIHO Y. 1983; Single amino acid substitution in 30K protein of TMV, defective in virus transport function. Virology 131:255–258
    [Google Scholar]
  16. PEDEN K. W. C., SYMONS R. H. 1973; Cucumber mosaic virus contains a functionally divided genome. Virology 53:487–492
    [Google Scholar]
  17. REZAIAN M. A., WILLIAMS R. H. V., GORDON K. H. J., GOULD A. R., SYMONS R. H. 1984; Nucleotide sequence of cucumber-mosaic-virus RNA 2 reveals a translation product significantly homologous to corresponding proteins of other viruses. European Journal of Biochemistry 143:277–284
    [Google Scholar]
  18. REZAIAN M. A., WILLIAMS R. H. V., SYMONS R. H. 1985; Nucleotide sequence of cucumber mosaic virus RNA 1 and homologies with other viral RNAs. European Journal of Biochemistry 150:331–339
    [Google Scholar]
  19. ROTH J., BENDAYAN M., ORCI L. 1978; Ultrastructural localization of intracellular antigens by the use of Protein A-gold complex. Journal of Histochemistry and Cytochemistry 26:1074–1081
    [Google Scholar]
  20. SAVITHRI H. S., MURTHY M. R. N. 1983; Evolutionary relationship of alfalfa mosaic virus with cucumber mosaic virus and brome mosaic virus. Journal of Biosciences 5:183–187
    [Google Scholar]
  21. SCHWINGHAMER M. W., SYMONS R. H. 1977; Translation of the four major RNA species of cucumber mosaic virus in plant and animal cell-free systems and in toad oocytes. Virology 79:88–108
    [Google Scholar]
  22. STUSSI-GARAUD C, GARAUD J.-C, BERNA A., GODEFROY-COLBURN T. 1987; In situ location of an alfalfa mosaic virus non-structural protein in plant cell walls: correlation with virus transport. Journal of General Virology 68:1779–1784
    [Google Scholar]
  23. TOMENIUS K., CLAPHAM D., MESHI T. 1987; Localization by immunogold cytochemistry of the virus-coded 30K protein in plasmodesmata of leaves infected with tobacco mosaic virus. Virology 160:363–371
    [Google Scholar]
  24. VAN PELT-HEERSCHAP H., VERBEEK H., HUISMAN M. J., LOESCH-FRIES L. S., VAN VLOTEN-DOTING L. 1987; Non-Structural proteins and RNAs of alfalfa mosaic virus synthesized in tobacco and cowpea protoplasts. Virology 161:190–197
    [Google Scholar]
  25. WATANABE Y., MORITA N., NISHLGUCHL M., OKADA Y. 1987; Attenuated strains of tobacco mosaic virus reduced synthesis of a viral protein with a cell-to-cell movement function. Journal of Molecular Biology 194:699–704
    [Google Scholar]
  26. ZIMMERN D. 1983; Homologous proteins encoded by yeast mitochondrial introns and by a group of RNA viruses from plants. Journal of Molecular Biology 171:345–352
    [Google Scholar]
  27. ZIMMERN D., HUNTER T. 1983; Point mutation in the 30K open reading frame of TMV implicated in temperature-sensitive assembly and local lesion spreading of mutant Ni 2519. EMBO Journal 2:1893–1900
    [Google Scholar]
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