1887

Abstract

To investigate the process of tubule formation for the cauliflower mosaic virus movement protein (CaMV MP), the green fluorescent protein (GFP) was fused to the MP to provide a vital marker for MP location after expression in insect cells. In contrast to the long tubular structures seen previously following baculovirus-based expression of the wild-type MP, the fusion protein produced only aggregates of fluorescing material in the cytoplasm. However, by co-expressing wild-type MP and GFP–MP, or by engineering their co-accumulation by introducing a foot-and-mouth disease virus 2A cleavage sequence between GFP and MP, long GFP-fluorescing tubules were formed. The experiments suggest that the presence of GFP at the N or C terminus of the tubule-forming domain of the CaMV MP places steric constraints upon the aggregation of the MP into a tubule but that this can be overcome by providing wild-type protein for inclusion in the aggregate.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-81-7-1851
2000-07-01
2021-07-27
Loading full text...

Full text loading...

/deliver/fulltext/jgv/81/7/0811851a.html?itemId=/content/journal/jgv/10.1099/0022-1317-81-7-1851&mimeType=html&fmt=ahah

References

  1. Crameri, A., Whitehorn, E. A., Tate, E. & Stemmer, W. P. C. (1996). Improved green fluorescent protein by molecular evolution using DNA shuffling. Nature Biotechnology 14, 315-319.[CrossRef] [Google Scholar]
  2. Fütterer, J., Bonneville, J. M. & Hohn, T. (1990). Cauliflower mosaic virus as a gene expression vector for plants. Physiologia Plantarum 79, 154-157.[CrossRef] [Google Scholar]
  3. Gardner, R. C., Howarth, A. J., Hahn, P., Brown-Luedi, M., Shepherd, R. J. & Messing, J. (1981). The complete nucleotide sequence of an infectious clone of cauliflower mosaic virus by M13mp7 shotgun sequencing. Nucleic Acids Research 9, 2871-2888.[CrossRef] [Google Scholar]
  4. Grieco, F., Castellano, M. A., Di Sansebastiano, G. P., Maggipinto, G., Neuhaus, J.-M. & Martelli, G. P. (1999). Subcellular localization and in vivo identification of the putative movement protein of olive latent virus 2. Journal of General Virology 80, 1103-1109. [Google Scholar]
  5. Harker, C. L., Mullineaux, P. M., Bryant, J. A. & Maule, A. J. (1987). Detection of CaMV gene 1 and gene VI protein products in vivo using antisera raised to COOH-terminal β-galactosidase proteins. Plant Molecular Biology 8, 275-287.[CrossRef] [Google Scholar]
  6. Haseloff, J., Siemering, K. R., Prasher, D. C. & Hodge, S. (1997). Removal of a cryptic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly. Proceedings of National Academy of Sciences, USA 94, 2122-2127.[CrossRef] [Google Scholar]
  7. Kasteel, D. T. J., Perbal, M.-C., Boyer, J.-C., Wellink, J., Goldbach, R. W., Maule, A. J. & van Lent, J. W. M. (1996). The movement proteins of cowpea mosaic virus and cauliflower mosaic virus induce tubular structures in plant and insect cells. Journal of General Virology 77, 2857-2864.[CrossRef] [Google Scholar]
  8. Kasteel, D. T. J., Wellink, J., Goldbach, R. W. & van Lent, J. W. M. (1997). Isolation and characterisation of tubular structures of cowpea mosaic virus. Journal of General Virology 78, 3167-3170. [Google Scholar]
  9. King, L. A. & Possee, R. D. (1992).The Baculovirus Expression System. A Laboratory Guide. London: Chapman & Hall.
  10. Kiss-László, Z., Blanc, S. & Hohn, T. (1995). Splicing of cauliflower mosaic virus 35S RNA is essential for viral infectivity. EMBO Journal 14, 3552-3562. [Google Scholar]
  11. Lekkerkerker, A., Wellink, J., Yuan, P., van Lent, J., Goldbach, R. W. & van Kammen, A. (1996). Distinct functional domains in the cowpea mosaic virus movement protein. Journal of Virology 70, 5658-5661. [Google Scholar]
  12. Maule, A. J., Usmany, M., Wilson, I. G., Boudazin, G. & Vlak, J. M. (1992). Biophysical and biochemical properties of baculovirus-expressed CaMV P1 protein.Virus Genes 6, 5-18.[CrossRef] [Google Scholar]
  13. Oparka, K. J., Boevink, P. & Santa Cruz, S. (1996). Studying the movement of plant viruses using green fluorescent protein. Trends in Plant Science 1, 412-418.[CrossRef] [Google Scholar]
  14. Perbal, M.-C., Thomas, C. L. & Maule, A. J. (1993). Cauliflower mosaic virus gene 1 product (P1) forms tubular structures which extend from the surface of infected protoplasts. Virology 195, 281-285.[CrossRef] [Google Scholar]
  15. Ryan, M. D. & Drew, J. (1994). Foot-and-mouth disease virus 2A oligopeptide mediated cleavage of an artificial polyprotein. EMBO Journal 13, 928-933. [Google Scholar]
  16. Ryan, M. D., King, A. M. Q. & Thomas, G. P. (1991). Cleavage of foot-and-mouth disease virus polyprotein is mediated by residues located within a 19 amino acid sequence.Journal of General Virology 72, 2727-2732.[CrossRef] [Google Scholar]
  17. Thomas, C. L. & Maule, A. J. (1995a). Identification of structural domains within the cauliflower mosaic virus movement protein by scanning deletion mutagenesis and epitope tagging. Plant Cell 7, 561-572.[CrossRef] [Google Scholar]
  18. Thomas, C. L. & Maule, A. J. (1995b). Identification of the cauliflower mosaic virus movement protein RNA binding domain. Virology 206, 1145-1149.[CrossRef] [Google Scholar]
  19. Thomas, C. L. & Maule, A. J. (1999). Identification of inhibitory mutants of cauliflower mosaic virus movement protein function after expression in insect cells. Journal of Virology 73, 7886-7890. [Google Scholar]
  20. Zheng, H., Wang, G. & Zhang, L. (1997). Alfalfa mosaic virus movement protein induces tubules in plant protoplasts. Molecular Plant–Microbe Interactions 10, 1010-1014.[CrossRef] [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-81-7-1851
Loading
/content/journal/jgv/10.1099/0022-1317-81-7-1851
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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