1887

Abstract

The coat protein of alfalfa mosaic virus (AMV) was cloned and expressed in as a fusion protein containing a 37 amino acid extension with a (His) region for affinity purification. About half of the expressed recombinant coat protein (rCP) was soluble upon extraction and half was insoluble in inclusion bodies. Western blot analysis confirmed the identity of the rCP and protoplast infectivity assays indicated that the rCP was biologically active in an early event of AMV infection, called genome activation. The rCP assembled into = 1 empty icosahedral particles, as described previously for native coat protein. Empty particles formed hexagonal crystals that diffracted X-rays to 5.5 Å resolution. The crystals of trypsin-treated particles of rCP appear to be isomorphous with crystals of trypsintreated particles of native coat protein. Spherical particles containing RNA assembled when the rCP was combined with transcripts of AMV RNA4, the smallest naturally encapsidated AMV RNA. Bacilliform particles that resembled native virions assembled when the rCP was combined with transcripts of RNA1, the largest genomic RNA.

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1996-04-01
2022-05-28
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References

  1. Allison R., Thompson C., Ahlquist P. 1990; Regeneration of a functional RNA virus genome by recombination between deletion mutants and requirement for cowpea chlorotic mottle virus 3a and coat genes for systemic infection. Proceedings of the National Academy of Sciences, USA 87:1820–1824
    [Google Scholar]
  2. Baer M. L., Houser F., Loesch-Fries L. S., Gehrke L. 1994; Specific RNA binding by amino-terminal peptides of alfalfa mosaic virus coat protein. EMBO Journal 13:727–735
    [Google Scholar]
  3. Bancroft J. B., Hiebert E. 1967; Formation of an infectious nucleoprotein from protein and nucleic acid isolated from a small spherical virus. Virology 32:354–356
    [Google Scholar]
  4. Bol J. F., Van Vloten-Doting L., Jaspars E. M. J. 1971; A functional equivalence of top component a RNA and coat protein in the initiation of infection by alfalfa mosaic virus. Virology 46:73–85
    [Google Scholar]
  5. Bol J. F., Kraal B., Brederode F. Th. 1974; Limited proteolysis of alfalfa mosaic virus: influence on the structural and biological function of the coat protein. Virology 58:101–110
    [Google Scholar]
  6. Chapman S., Hills G., Watts J., Baulcombe D. 1992; Mutational analysis of the coat protein gene of potato virus X: effects on virion morphology and viral pathogenicity. Virology 191:223–230
    [Google Scholar]
  7. De Graaff M., Man in’t Weld M. R., Jaspars E. M. J. 1995; In vitro evidence that the coat protein of alfalfa mosaic virus plays a direct role in the regulation of plus and minus RNA synthesis: implications for the life cycle of alfalfa mosaic virus. Virology 208:583–589
    [Google Scholar]
  8. Dolja V. V., Haldeman R., Robertson N. L., Dougherty W. G., Carrington J. C. 1994; Distinct functions of capsid protein in assembly and movement of tobacco etch potyvirus in plants. EMBO Journal 13:1482–1491
    [Google Scholar]
  9. Driedonks R. A., Krijgsman P. C. J., Mellema J. E. 1977; Alfalfa mosaic virus protein polymerization. Journal of Molecular Biology 113:123–140
    [Google Scholar]
  10. Driedonks R. A., Krijgsman P. C. J., Mellema J. E. 1978; A characterization of alfalfa-mosaic-virus protein polymerization in the presence of nucleic acid. European Journal of Biochemistry 82:405–417
    [Google Scholar]
  11. Fukuyama K., Abdel-Meguid S. S., Rossmann M. G. 1981; Crystallization of alfalfa mosaic virus coat protein as a T = 1 aggregate. Journal of Molecular Biology 150:33–41
    [Google Scholar]
  12. Fukuyama K., Abdel-Meguid S. S., Rossmann M. G. 1983; Structure of a T = 1 aggregate of alfalfa mosaic virus coat protein seen at 4.5 Å resolution. Journal of Molecular Biology 167:873–894
    [Google Scholar]
  13. Halk E. L. 1981; Comparative electrophoretic mobility of three ilarviruses and alfalfa mosaic virus in polyacrylamide gels. Abstract, Fifth International Congress of Virology, P1/13
    [Google Scholar]
  14. Halk E. L. 1986; Serotyping plant viruses with monoclonal antibodies. Methods in Enzymology 118:766–780
    [Google Scholar]
  15. Hull R. 1970; Studies on alfalfa mosaic virus. III. Reversible dissociation and reconstitution studies. Virology 40:34–47
    [Google Scholar]
  16. Jaspars E. M. J. 1985; Interaction of alfalfa mosaic virus nucleic acid and protein. In Molecular Plant Virology vol 1 pp 155–221 Edited by Davies J. W. Boca Raton, Fla.: CRC Press;
    [Google Scholar]
  17. Kruseman J., Kraal B., Jaspars E. M. J., Bol J. F., Brederode F. Th., Veldstra H. 1971; Molecular weight of the coat protein of alfalfa mosaic virus. Biochemistry 10:447–455
    [Google Scholar]
  18. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  19. Lebeurier G., Fraenkel-Conrat H., Wurtz M., Hirth L. 1971; Self-assembly of protein subunits from alfalfa mosaic virus. Virology 43:51–61
    [Google Scholar]
  20. Loesch-Fries L. S., Hall T. C. 1980; Synthesis, accumulation and encapsidation of individual brome mosaic virus RNA components in barley protoplasts. Journal of General Virology 47:323–332
    [Google Scholar]
  21. Loesch-Fries L. S., Jarvis N. P., Krahn K. J., Nelson S. E., Hall T. C. 1985; Expression of alfalfa mosaic virus RNA 4 cDNA transcripts in vitro and in vivo . Virology 146:177–187
    [Google Scholar]
  22. Loesch-Fries L. S., Merlo D., Zinnen T., Burhop L., Hill K., Krahn K., Jarvis N., Nelson S., Halk E. 1987; Expression of alfalfa mosaic virus RNA 4 in transgenic plants confers virus resistance. EMBO Journal 6:1845–1851
    [Google Scholar]
  23. Marsh L. E., Huntley C. C., Pogue G. P., Connell J. P., Hall T. C. 1991; Regulation of (+):(—) strand asymmetry in replication of brome mosaic virus RNA. Virology 182:76–83
    [Google Scholar]
  24. Otwinowski Z. 1993; Oscillation data reduction program. In Proceedings of the CCP4 Study Weekend: Data Collection and Processing pp 55–62 Edited by Sawyer L., Isaacs N., Bailey S. SERC Daresbury Laboratory; UK:
    [Google Scholar]
  25. Quadt R., Rosdorff H. J. M., Hunt T. W., Jaspars E. M. J. 1991; Analysis of protein composition of alfalfa mosaic virus RNA-dependent RNA polymerase. Virology 182:309–315
    [Google Scholar]
  26. Reusken C. B. E. M., Neeleman L., Bol J. F. 1994; The 3′-untranslated region of alfalfa mosaic virus RNA3 contains at least two independent binding sites for viral coat protein. Nucleic Acids Research 22:1346–1353
    [Google Scholar]
  27. Sacher R., Ahlquist P. 1989; Effects of deletions in the N-terminal basic arm of brome mosaic virus coat protein on RNA packaging and systemic infection. Journal of Virology 63:4545–4552
    [Google Scholar]
  28. Samac D. A., Nelson S. E., Loesch-Fries L. S. 1983; Virus protein synthesis in alfalfa mosaic virus infected alfalfa protoplasts. Virology 131:455–462
    [Google Scholar]
  29. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual 2nd edn New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  30. Sehnke P. C., Johnson J. E. 1994; A chromatographic analysis of capsid protein isolated from alfalfa mosaic virus: zinc binding and proteolysis cause distinct charge heterogeneity. Virology 204:843–846
    [Google Scholar]
  31. Van der Kuyl A. C., Neeleman L., Bol J. F. 1991a; Complementation and recombination between alfalfa mosaic virus RNA3 mutants in tobacco plants. Virology 183:731–738
    [Google Scholar]
  32. Van der Kuyl A. C., Neeleman L., Bol J. F. 1991b; Role of alfalfa mosaic virus coat protein in regulation of the balance between viral plus and minus strand RNA synthesis. Virology 185:496–499
    [Google Scholar]
  33. Van der Vossen E. A. G., Neeleman L., Bol J. F. 1994; Early and late functions of alfalfa mosaic virus coat protein can be mutated separately. Virology 202:891–903
    [Google Scholar]
  34. Yusibov V. M., Loesch-Fries L. S. 1995; N-terminal basic amino acids of alfalfa mosaic virus coat protein involved in the initiation of infection. Virology 208:405–407
    [Google Scholar]
  35. Zhao X., Fox J. M., Olson N. H., Baker T. S., Young M. J. 1995; In vitro assembly of cowpea chlorotic mottle virus from coat protein expressed in Escherichia coli and in w′zro-transcribed viral cDNA. Virology 207:486–494
    [Google Scholar]
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