1887

Abstract

Recombinant DNA molecules containing cDNA to a soybean mosaic virus (SMV) RNA genome were constructed and partial nucleotide sequences determined for two cDNA inserts, pSMV-34 and pSMV-35. Comparison of the predicted amino acid sequence encoded by the pSMV-34 cDNA insert to other potyvirus protein sequences revealed extensive homology with the region of the genome encoding the NIa proteinase, with conservation of the amino acids proposed to form the catalytic triad of the active site. Cell-free transcription and translation of the cloned cDNA sequence containing the NIa open reading frame and flanking sequences revealed that NIa proteinase sequences, which were expressed as part of a high precursor, were able to undergo proteolytic processing. Alteration of the codon for one of the putative active site residues by site-directed mutagenesis eliminated processing and resulted in the accumulation of a high precursor. Based on predicted amino acid sequences at five putative cleavage sites within the SMV polyprotein, a consensus SMV NIa proteinase cleavage sequence of Glu/Asn- Xaa-Val-Xaa-Xaa-Gln ↓ Gly/Ser was proposed. The SMV NIa proteinase and its putative cleavage sites maintained motifs found in other potyviruses.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-71-9-1921
1990-09-01
2022-08-11
Loading full text...

Full text loading...

/deliver/fulltext/jgv/71/9/JV0710091921.html?itemId=/content/journal/jgv/10.1099/0022-1317-71-9-1921&mimeType=html&fmt=ahah

References

  1. Allison R. F., 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. Argos P., Kamer G., Nicklin M. J. H., Wimmer E. 1984; Similarity in gene organization and homology between proteins of animal picomaviruses and a plant comovirus suggest common ancestry of these virus families. Nucleic Acids Research 12:7251–7267
    [Google Scholar]
  3. Bazan J. F., Fletterick R. J. 1988; Viral cysteine proteases are homologous to the trypsin-like family of serine proteases: structural and functional implications. Proceedings of the National Academy of Sciences, U.S.A 85:7872–7876
    [Google Scholar]
  4. Bos L. 1972; Soybean mosaic virus. CMI/AAB Descriptions of Plant Viruses93
    [Google Scholar]
  5. Calvert L., Ghabrial S. A. 1983; Enhancement by soybean mosaic virus of bean pod mottle virus titer in doubly infected soybean. Phytopathology 78:992–997
    [Google Scholar]
  6. Carrington J. C., Dougherty W. G. 1987a; Small nuclear inclusion protein encoded by a plant potyvirus genome is a protease. Journal of Virology 61:2540–2548
    [Google Scholar]
  7. Carrington J. C., Dougherty W. G. 1987b; Processing of the tobacco etch virus 49K protease requires autoprocessing. Virology 160:355–362
    [Google Scholar]
  8. Carrington J. C., Parks T. D., Cary S. M., Dougherty W. G. 1987; Vectors for cell-free expression and mutagenesis of protein coding sequences. Nucleic Acids Research 15:10066
    [Google Scholar]
  9. Carrington J. C., Cary S. M., Parks T. D., Dougherty W. G. 1989; A second proteinase encoded by a plant potyvirus genome. EMBO Journal 8:365–370
    [Google Scholar]
  10. Cho E. K., Goodman R. M. 1979; Strains of soybean mosaic virus: classification based on virulence in resistant soybean cultivars. Phytopathology 69:467–470
    [Google Scholar]
  11. Domier L. L., Franklin K. M., Shahabuddin M., Hellmann G. M., Overmeyer J. H., Hiremath S. T., Siaw M. F., 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]
  12. Dougherty W. G., Carrington J. C. 1988; Expression and function of poty viral gene products. Annual Review of Phytopathology 26:123–143
    [Google Scholar]
  13. Dougherty W. G., Parks T. D. 1989; Molecular genetic and biochemical evidence for the involvement of the heptapeptide cleavage sequence in determining the reaction profile at two tobacco etch virus cleavage sites in cell-free assays. Virology 172:145–155
    [Google Scholar]
  14. Dougherty W. G., Parks T. D., Cary S. M., Bazan J. F., Fletterick R. J. 1989; Characterization of the catalytic residues of the tobacco etch virus 49-kDa proteinase. Virology 172:302–310
    [Google Scholar]
  15. Eggenberger A. L., Stark D. M., Beachy R. N. 1989; The nucleotide sequence of a soybean mosaic virus coat protein-coding region and its expression in Escherichia coli, Agrobacterium tumefaciens and tobacco callus. Journal of General Virology 70:1853–1860
    [Google Scholar]
  16. Garcia J. A., Riechmann J. L., Lain S. 1989; Proteolytic activity of the plumpox potyvirus NIa-like protein in Escherichia coli . Virology 170:362–369
    [Google Scholar]
  17. Goldbach R. W. 1987; Genome similarities between plant and animal RNA viruses. Microbiological Sciences 4:197–202
    [Google Scholar]
  18. Gubler U., Hoffman B. J. 1983; A simple and very efficient method for generating cDNA libraries. Gene 25:263–269
    [Google Scholar]
  19. Hellen C. U. T., Kräusslich H. G., Wimmer E. 1989; Proteolytic processing of polyproteins in the replication of RNA viruses. Biochemistry 28:9881–9890
    [Google Scholar]
  20. Hellmann G. M., Shaw J. G., Lesnaw J. A., Chu L.-Y., Pirone T. P., Rhoads R. E. 1980; Cell-free translation of tobacco vein mottling virus RNA. Virology 106:206–216
    [Google Scholar]
  21. Hellmann G. M., Shaw J. G., Rhoads R. E. 1988; In vitro analysis of tobacco vein mottling virus NIa cistron: evidence for a virus-encoded protease. Virology 163:554–562
    [Google Scholar]
  22. Kräusslich H. G., Wimmer E. 1988 Viral proteinoses. Annual Review of Biochemistry 57:701–754
    [Google Scholar]
  23. Maiss E., Timpe U., Brisske A., Jelkmann W., Casper R., Himmler G., Mattanovich D., Katinger H. W. D. 1989; The complete nucleotide sequence of plum pox virus RNA. Journal of General Virology 70:513–524
    [Google Scholar]
  24. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  25. Robaglia C., Durand-Tardif M., Tronchet M., Boudazin G., Astier-Manifacier S., Casse-Delbart F. 1989; Nucleotide sequence of potato virus Y (N strain) genomic RNA. Journal of General Virology 70:935–947
    [Google Scholar]
  26. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, U.S.A 74:5463–5467
    [Google Scholar]
  27. Sinclair J. B., Backman P. A. 1989 Compendium of Soybean Diseases, 3rd edn. St Paul: American Phytopathological Society;
    [Google Scholar]
  28. Strauss J. H., Strauss E. G. 1988; Evolution of RNA viruses. Annual Review of Microbiology 42:657–683
    [Google Scholar]
  29. Taylor J. W., Ott J., Eckstein F. 1985a; The rapid generation of oligonucleotide-directed mutations at high frequency using phosphorothioate-modified DNA. Nucleic Acids Research 13:8765–8785
    [Google Scholar]
  30. Taylor J. W., Schmidt W., Cosstick R., Okruszek A., Eckstein F. 1985b; The use of phosphorothioate-modified DNA in restriction enzyme reactions to prepare nicked DNA. Nucleic Acids Research 13:8749–8764
    [Google Scholar]
  31. Vance V. B., Beachy R. N. 1984a; Translation of soybean mosaic virus RNA in vitro: evidence of protein processing. Virology 132:271–281
    [Google Scholar]
  32. Vance V. B., Beachy R. N. 1984b; Detection of genomic-length soybean mosaic virus RNA on polyribosomes of infected soybean leaves. Virology 138:26–36
    [Google Scholar]
  33. Wellink J., van Kammen A. 1988; Proteases involved in the processing of viral polyproteins. Archives of Virology 98:1–26
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-71-9-1921
Loading
/content/journal/jgv/10.1099/0022-1317-71-9-1921
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