1887

Abstract

Mutants of tobacco vein mottling virus (TVMV) with substitutions of Lys or Arg for Asp in the DAG motif at position 5 in the coat protein (CP) failed to infect tobacco plants systemically, but replicated and produced virions in protoplasts. Occasional systemic infections occurred when or transgenic tobacco plantsexpressing wild- type TVMV CP were inoculated with these mutants, but viral progeny contained reversions to negatively or non-charged amino acids at position 5 or substitutions of Glu for Lys at position 8. The compensatory nature of these mutations was demonstrated by recreating one of the most common alterations. Tobacco etch virus (TEV) mutants with substitutions of Lys for Asp in the two DAG motifs near the CP N terminus also failed to infect tobacco plants systemically, and histochemical analysis showed limited movement. A certain net charge evidently must be maintained near the CP N terminus for systemic movement to occur.

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1998-01-01
2022-05-28
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References

  1. Allison R., Johnston R. E., Dougherty W. G. 1986; The nucleotide sequence of the coding region of tobacco etch virus genomic RNA : evidence for the synthesis of a single polyprotein. Virology 154:9–20
    [Google Scholar]
  2. Atreya C. D., Pirone T. P. 1993; Mutational analysis of the helper component proteinase gene of a potyvirus: effects of amino acid substitutions, deletions, and gene replacement on virulence and aphid transmissibility. Proceedings of the National Academy of Sciences, USA 90:11919–11923
    [Google Scholar]
  3. Atreya P. L., Lopez-Moya J. J., Chu M., Atreya C. D., Pirone T. P. 1995; Mutational analysis of the coat protein N-terminal amino acids residues involved in potyvirus transmission by aphids. Journal of General Virology 76:265–270
    [Google Scholar]
  4. Carrington J. C., Haldeman R., Dolja V. V., Restrepo-Hartwig M. A. 1993; Internal cleavage and frans-proteolytic activities of the VPg-proteinase (NIa) of tobacco etch potyvirus in vivo. Journal of Virology 67:6995–7000
    [Google Scholar]
  5. Carrington J. C., Kasschau K. D., Mahajan S. K., Schaad M. C. 1996; Cell-to-cell and long-distance transport of viruses in plants. Plant Cell 8:1669–1681
    [Google Scholar]
  6. Cronin S., Verchot J., Haldeman-Cahill R., Schaad M. C., Carrington J. C. 1995; Long-distance movement factor: a transport function of the potyvirus helper component proteinase. Plant Cell 7:549–559
    [Google Scholar]
  7. Dolja V. V., McBride H. J., Carrington J. C. 1992; Tagging of plant potyvirus replication and movement by insertion of β-glucuronidase into the viral polyprotein. Proceedings of the National Academy of Sciences, USA 89:10208–10212
    [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. Dolja V. V., Haldeman-Cahill R., Montgomery A. E., Vandenbosch K. A., Carrington J. C. 1995; Capsid protein determinants involved in cell-to-cell and long distance movement of tobacco etch potyvirus. Virology 206:1007–1016
    [Google Scholar]
  10. Domier L. L., Franklin K. M., Shahabuddin M., Hellmann G. M., Overmeyer J. H., Hiremath S. T., Siaw M. F. E., Lomonossoff G. P., Shaw J. G., Rhoads R. E. 1986; The nucleotide sequence of tobacco vein mottling virus RNA. Nucleic Acids Research 14:5417–5430
    [Google Scholar]
  11. Domier L. L., Franklin K. M., Hunt A. G., Rhoads R. E., Shaw J. G. 1989; Infectious in vitro transcripts from cloned cDNA of a potyvirus, tobacco vein mottling virus. Proceedings of the National Academy of Sciences, USA 86:3509–3513
    [Google Scholar]
  12. Hong Y., Levay K., Murphy J. F., Klein P. G., Shaw J. G., Hunt A. G. 1995; A potyvirus polymerase interacts with the viral coat protein and VPg in yeast cells. Virology 214:159–166
    [Google Scholar]
  13. Kasschau K. D., Cronin S., Carrington J. C. 1997; Genome amplification and long-distance movement functions associated with the central domain of tobacco etch virus helper-component-proteinase. Virology 228:251–262
    [Google Scholar]
  14. Kunkel T. A., Roberts J. D., Zakour R. A. 1987; Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods in Enzymology 154:367–382
    [Google Scholar]
  15. Luciano C. S., Rhoads R. E., Shaw J. G. 1987; Synthesis of potyviral RNA and proteins in tobacco mesophyll protoplasts inoculated by electroporation. Plant Science 51:295–303
    [Google Scholar]
  16. Otsuki Y., Takebe I., Honda Y., Matsui C. 1972; Ultrastructure of infection of tobacco mesophyll protoplasts by tobacco mosaic virus. Virology 49:188–194
    [Google Scholar]
  17. Shukla D. D., Ward C. W. 1989; Structure of potyvirus coat protein and its application in the taxonomy of the potyvirus group. Advances in Virus Research 36:273–314
    [Google Scholar]
  18. Tao B. Y., Lee K. C. P. 1994; Mutagenesis by PCR. In PCR Technology: Current Innovations pp. 69–83 Griffin H. G., Griffin A. M. Edited by Boca Raton: CRC Press;
    [Google Scholar]
  19. Wang R. Y., Ammar E. D., Thornbury D. W., Lopez-Moya J. J., Pirone T. P. 1996; Loss of potyvirus transmissibility and helper-component activity correlate with non-retention of virions in aphid stylets. Journal of General Virology 77:861–867
    [Google Scholar]
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