1887

Abstract

The proteinase of (genus ) is responsible for proteolytic cleavage of the RNA2-encoded polyprotein (P2) at two cleavage sites, allowing definition of the domains for the movement protein (MP) and coat protein. In this study, we have characterized a third cleavage site in the N-terminal region of P2 using an processing assay and partial cDNA clones. Results from site-directed mutagenesis of putative cleavage sites suggest that cleavage occurs at dipeptide Q/G. Cleavage at this site is predicted to result in the release of two proteins from the N-terminal region of P2: a 34 kDa protein located at the N terminus of P2 (assuming translation initiation at the first AUG codon) and a 71 kDa protein located immediately upstream of the MP domain. In contrast, only one protein domain is present in the equivalent region of the P2 polyprotein of other characterized nepoviruses.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-82-7-1785
2001-07-01
2024-05-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/82/7/0821785a.html?itemId=/content/journal/jgv/10.1099/0022-1317-82-7-1785&mimeType=html&fmt=ahah

References

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. 1990; Basic local alignment search tool. Journal of Molecular Biology 215:403–410
    [Google Scholar]
  2. Carrier K., Hans F., Sanfaçon H. 1999; Mutagenesis of amino acids at two tomato ringspot nepovirus cleavage sites: effect on proteolytic processing in cis and in trans by the 3C-like protease. Virology 258:161–175
    [Google Scholar]
  3. Clark A. J., Bertens P., Wellink J., Shanks M., Lomonossoff G. P. 1999; Studies on hybrid comoviruses reveal the importance of three-dimensional structure for processing of the viral coat protein and show that the specificity of cleavage is greater in trans than in cis. Virology 263:184–194
    [Google Scholar]
  4. Demangeat G., Hemmer O., Fritsch C., Le Gall O., Candresse T. 1991; In vitro processing of the RNA-2-encoded polyprotein of two nepoviruses: tomato black ring virus and grapevine chrome mosaic virus. Journal of General Virology 72:247–252
    [Google Scholar]
  5. Gaire F., Schmitt C., Stussi-Garaud C., Pinck L., Ritzenthaler C. 1999; Protein 2A of grapevine fanleaf nepovirus is implicated in RNA2 replication and colocalizes to the replication site. Virology 264:25–36
    [Google Scholar]
  6. Guermeur Y., Geourjon C., Gallinari P., Deleage G. 1999; Improved performance in protein secondary structure prediction by inhomogeneous score combination. Bioinformatics 14:413–421
    [Google Scholar]
  7. Hans F., Sanfaçon H. 1995; Tomato ringspot nepovirus protease: characterization and cleavage site specificity. Journal of General Virology 76:917–927
    [Google Scholar]
  8. Hibrand L., Le Gall O., Candresse T., Dunez J. 1992; Immunodetection of the proteins encoded by grapevine chrome mosaic nepovirus RNA2. Journal of General Virology 73:2093–2098
    [Google Scholar]
  9. Latvala S., Susi P., Kalkkinen N., Lehto K. 1998; Characterization of the coat protein gene of mite-transmitted blackcurrant reversion associated nepovirus. Virus Research 53:1–11
    [Google Scholar]
  10. Latvala-Kilby S., Lehto K. 1999; The complete nucleotide sequence of RNA2 of blackcurrant reversion nepovirus. Virus Research 65:87–92
    [Google Scholar]
  11. Margis R., Ritzenthaler C., Reinbolt J., Pinck M., Pinck L. 1993; Genome organization of grapevine fanleaf nepovirus RNA2 deduced from the 122K polyprotein P2 in vitro cleavage products. Journal of General Virology 74:1919–1926
    [Google Scholar]
  12. Mayo M. A., Robinson D. J. 1996; Nepoviruses: molecular biology and replication. In The Plant Viruses pp 139–185 Edited by Harrison B. D., Murant A. F. New York: Plenum Press;
    [Google Scholar]
  13. Rott M. E., Tremaine J. H., Rochon D. M. 1991a; Nucleotide sequence of tomato ringspot virus RNA-2. Journal of General Virology 76:465–473
    [Google Scholar]
  14. Rott M. E., Tremaine J. H., Rochon D. M. 1991b; Comparison of the 5′ and 3′ termini of tomato ringspot virus RNA1 and RNA2: evidence for RNA recombination. Virology 185:468–472
    [Google Scholar]
  15. Sanfaçon H. 1995; Nepovirus. In Pathogenesis and Host Specificity in Plant Disease: Histopathological Biochemical & Molecular Bases , vol. III, Viruses & Viroids pp 129–141 Edited by Singh R. P., Singh U. S., Kohmoto K. Oxford: Pergamon Press;
    [Google Scholar]
  16. Van Bokhoven H., Le Gall O., Kasteel D., Verver J., Wellink J., Van Kammen A. 1993; Cis - and trans- acting elements in cowpea mosaic virus RNA replication. Virology 195:377–386
    [Google Scholar]
  17. Wang A., Sanfaçon H. 2000; Proteolytic processing at a novel cleavage site in the N-terminal region of the tomato ringspot nepovirus RNA1-encoded polyprotein in vitro . Journal of General Virology 81:2771–2781
    [Google Scholar]
  18. Wang A., Carrier K., Chisholm J., Wieczorek A., Huguenot C., Sanfaçon H. 1999; Proteolytic processing of tomato ringspot nepovirus 3C-like protease precursors: definition of the domains for the VPg, protease and putative RNA-dependent RNA polymerase. Journal of General Virology 80:799–809
    [Google Scholar]
  19. Ypma-Wong M. F., Filman D. J., Hogle J. M., Semler B. L. 1988; Structural domains of the poliovirus polyprotein are major determinants for proteolytic processing at Gln–Gly pairs. Journal of Biological Chemistry 263:17846–17856
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-82-7-1785
Loading
/content/journal/jgv/10.1099/0022-1317-82-7-1785
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