1887

Abstract

In order to elucidate the function of the C-terminal region of cowpea chlorotic mottle bromovirus (CCMV) movement protein (MP) in cell-to-cell movement, a set of deletions ranging from 10 to 80 amino acids (deltaMP10, deltaMP20, deltaMP33, deltaMP43, deltaMP60 and deltaMP80) was engineered into the MP gene encoded by the biologically active clone C3/deltaCP-EGFP, a variant of CCMV RNA3 that contained wild-type (wt) MP and the enhanced green fluorescent protein (EGFP) gene in place of the coat protein (CP). The effect of each MP deletion on cell-to-cell movement was examined in three susceptible host plants: Chenopodium quinoa, Nicotiana benthamiana and cowpea (Vigno sinensis cv. Black Eye). The results indicate that, except for mutant deltaMP43, infections resulting from the deletion mutants remained subliminal. Interestingly, infections resulting from inoculating mutant deltaMP43, which lacked the 43 most C-terminal amino acids, spread rapidly between cells and the number of infected cells expressing EGFP approached that of control inoculations made with C3/deltaCP-EGFP. To verify whether the presence of wt CP altered the movement behaviour of these mutants, each MP deletion was also incorporated into the genetic background of wt CCMV RNA3 (pCC3) and inoculated independently to all three hosts. The results suggest that the overall movement process exhibited by each MP mutant is influenced profoundly by the presence of CP and the particular host plant tested.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-80-6-1357
1999-06-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/jgv/80/6/0801357a.html?itemId=/content/journal/jgv/10.1099/0022-1317-80-6-1357&mimeType=html&fmt=ahah

References

  1. Ahlquist P. 1994; Bromoviruses. In Encyclopedia of Virology vol 1 pp 181–185 Edited by Webster R. G., Granoff A. San Diego: Academic Press;
    [Google Scholar]
  2. Allison R. F., Janda M., Ahlquist P. 1988; Infectious in vitro transcripts from cowpea chlorotic mottle virus cDNA clones and exchange of individual RNA components with brome mosaic virus. Journal of Virology 62:3581–3588
    [Google Scholar]
  3. Allison R. F., 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]
  4. Carrington J. C., Kasschau K. D., Mahajan S. K., Schaad M. 1996; Cell-to-cell and long distance transport of viruses in plants. Plant Cell 8:1669–1681
    [Google Scholar]
  5. De Jong W., Mise K., Chu A., Ahlquist P. 1997; Effects of coat protein mutations and reduced movement protein expression on infection spread by cowpea chlorotic mottle virus and its hybrid derivatives. Virology 232:167–173
    [Google Scholar]
  6. Deom C. M., Lapidot M., Beachy R. N. 1992; Plant virus movement proteins. Cell 69:221–224
    [Google Scholar]
  7. Dreher T. W., Rao A. L. N., Hall T. C. 1989; Replication in vivo of mutant brome mosaic virus RNAs defective in aminoacylation. Journal of Molecular Biology 206:425–438
    [Google Scholar]
  8. Fujita Y., Mise K., Okuno T., Ahlquist P., Furusawa I. 1996; A single codon change in a conserved motif of a bromovirus movement protein gene confers compatibility with a new host. Virology 223:283–291
    [Google Scholar]
  9. Fujita Y., Mise K., Kajiura Y., Dohi K., Furusawa I. 1998; Nucleic acid-binding properties and subcellular localization of the 3a protein of brome mosaic bromovirus. Journal of General Virology 79:1273–1280
    [Google Scholar]
  10. Jansen K. A., Wolfs C. J., Lohuis H., Goldbach R. W., Verduin B. J. 1998; Characterization of the brome mosaic virus movement protein expressed in E. coli. Virology 242:387–394
    [Google Scholar]
  11. Kasteel D. T. J., van der Wel N. N., Jansen K. A. J., Goldbach R. W., van Lent J. W. M. 1997; Tubule-forming capacity of the movement proteins of alfalfa mosaic virus and brome mosaic virus. Journal of General Virology 78:2089–2093
    [Google Scholar]
  12. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  13. Mise K., Ahlquist P. 1995; Host-specificity restriction by bromo-virus cell-to-cell movement protein occurs after initial cell-to-cell spread of infection in nonhost plants. Virology 206:276–286
    [Google Scholar]
  14. Mise K., Allison R. F., Janda M., Ahlquist P. 1993; Bromovirus movement protein genes play a crucial role in host specificity. Journal of Virology 67:2815–2823
    [Google Scholar]
  15. Nagano H., Okuno T., Mise K., Furusawa I. 1997; Deletion of the C-terminal 33 amino acids of cucumber mosaic virus movement protein enables a chimeric brome mosaic virus to move from cell to cell. Journal of Virology 71:2270–2276
    [Google Scholar]
  16. Oparka K. J., Roberts A. G., Prior D. A. M., Chapman S., Baulcombe D. C., Santa Cruz S. 1995; Imaging the green fluorescent protein in plants-viruses carry the torch. Protoplasma 189:133–141
    [Google Scholar]
  17. Osman F., Grantham G. L., Rao A. L. N. 1997; Molecular studies on bromovirus capsid protein. IV. Coat protein exchanges between brome mosaic and cowpea chlorotic mottle viruses exhibit neutral effects in heterologous hosts. Virology 238:452–459
    [Google Scholar]
  18. Rao A. L. N. 1997; Molecular studies on bromovirus capsid protein. III. Analysis of cell-to-cell movement competence of coat protein defective variants of cowpea chlorotic mottle virus. Virology 232:385–395
    [Google Scholar]
  19. Rao A. L. N., Grantham G. L. 1995a; A spontaneous mutation in the movement protein gene of brome mosaic virus modulates symptom phenotype in Nicotiana benthamiana. Journal of Virology 69:2689–2691
    [Google Scholar]
  20. Rao A. L. N., Grantham G. L. 1995b; Biological significance of the seven amino-terminal basic residues of brome mosaic virus coat protein. Virology 211:42–52
    [Google Scholar]
  21. Rao A. L. N., Grantham G. L. 1996; Molecular studies on bromovirus capsid protein. II. Functional analysis of the amino-terminal arginine-rich motif and its role in encapsidation, movement, and pathology. Virology 226:294–305
    [Google Scholar]
  22. Rao A. L. N., Duggal R., Lahser F., Hall T. C. 1994; Analysis of RNA replication in plant viruses. In Methods in Molecular GeneticsMolecular Virology Techniques vol 4 pp 216–236 Edited by Adolph K. W. Orlando: Academic Press;
    [Google Scholar]
  23. Rao A. L. N., Cooper B., Deom C. M. 1998; Defective movement of viruses in the family Bromoviridae is differentially complemented in Nicotiana benthamiana expressing tobamovirus or dianthovirus movement proteins. Phytopathology 88:666–672
    [Google Scholar]
  24. Schmitz I., Rao A. L. N. 1996; Molecular studies on bromovirus capsid protein. I. Characterization of cell-to-cell movement-defective RNA3 variants of brome mosaic virus. Virology 226:281–293
    [Google Scholar]
  25. Traynor P., Young B. M., Ahlquist P. 1991; Deletion analysis of brome mosaic virus 2a protein: effects on RNA replication and systemic spread. Journal of Virology 65:2807–2815
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-80-6-1357
Loading
/content/journal/jgv/10.1099/0022-1317-80-6-1357
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error