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Volume 78, Issue 1

The cleavage activities of aphthovirus and cardiovirus 2A proteins. Free

Abstract

The primary 2A/2B polyprotein cleavage of aphtho- and cardioviruses is mediated by their 2A proteins cleaving C-terminally. Whilst the aphthovirus 2A region is only 16 aa (possibly 18 aa) long, the cardiovirus 2A protein is some 150 aa. We have previously shown that foot-and-mouth disease virus (FMDV) 2A is able to mediate cleavage in an artificial (chloramphenicol acetyltransferase/FMDV 2A/-glucuronidase [CAT-2A-GUS]) polyprotein system devoid of any other FMDV sequences with high (~85%), although not complete, cleavage. In this paper we show that insertion of upstream FMDV capsid protein 1D sequences increases the activity. In addition, we have demonstrated that the cardiovirus Theiler’s murine encephalomyelitis virus (TME) 2A protein, when linked to GUS in a single ORF, is able to cleave at its own C terminus with high efficiency - if not completely. The C-terminal 19 aa of TME 2A, together with the N-terminal proline residue of protein 2B, were inserted into the CAT/GUS artificial polyprotein system (in a single ORF). This recombinant [CAT-ΔTME2A-GUS] polyprotein was able to mediate cleavage with high (~85%) efficiency-directly comparable to the activity observed when FMDV 2A was inserted. A similar insertion into [CAT-GUS] of the C-terminal 19 aa of the cardiovirus encephalomyocarditis virus (EMC) 2A, together with the N-terminal proline residue of protein 2B, produced a [CAT- EMC2A- GUS] polyprotein which also mediated cleavage at ~85%. Analysis of the products of expression of these artificial polyproteins in a prokaryotic translation system did not, apparently, reveal any GUS cleavage product.

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© Society for General Microbiology Ltd 1997
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1997-01-01
2024-04-19
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References

  1. Allaire M., Chernaia M. M., Malcolm B. A., James M. N. G. 1994; Picornaviral 3C cysteine proteinases have a fold similar to chymotrypsin- like serine proteinases. Nature 369:72–76
     [Google Scholar]
  2. Bachrach H. L., Swaney J. B., vande Woude G. F. 1973; Isolation of the structural polypeptides of foot-and-mouth disease virus and analysis of their C-terminal sequences. Virology 52:520–528
     [Google Scholar]
  3. Batson S., Rundell K. 1991; Proteolysis at the 2A/2B junction in Theiler’s murine encephalomyelitis virus. Virology 181:764–767
     [Google Scholar]
  4. Bazan J. F., Fletterick R. J. 1988; Viral cysteine proteases are homologous to the trypsin-like serine proteases: structural and functional implications. Proceedings of the National Academy of Sciences USA: 857872–7876
     [Google Scholar]
  5. Carroll A. R., Rowlands D. J., Clarke B. E. 1984; The complete nucleotide sequence of the RNA coding for the primary translation product of foot and mouth disease virus. Nucleic Acids Research 12:2461–2472
     [Google Scholar]
  6. Gorbalenya A. E., Blinov V. M., Donchenko A. M. 1986; Poliovirus-encoded proteinase 3C : a possible evolutionary link between cellular serine and cysteine proteinase families. FEBS Letters 194:253–257
     [Google Scholar]
  7. Kurz C., Forss S., Kupper H., Strohmaier K., Schaller H. 1981; Nucleotide sequence and corresponding amino acid sequence of the gene for the major antigen of foot-and-mouth disease virus. Nucleic Acids Research 9:1919–1931
     [Google Scholar]
  8. Law K. M., Brown T. D. K. 1990; The complete nucleotide sequence of the GDVII strain of Theiler's murine encephalomyelitis virus (TMEV). Nucleic Acids Research 18:6707–6708
     [Google Scholar]
  9. Li F., Browning G. F., Studdert M. J., Crabb B. S. 1996; Equine rhinovirus 1 is more closely related to foot-and-mouth disease virus than to other picornaviruses. Proceedings of the National Academy of Sciences USA: 93990–995
     [Google Scholar]
  10. McKeown B. 1996 A study of the 2A and 3C mediated cleavage events in the processing of the foot-and-mouth disease virus polyprotein PhD thesis Queen’s University, Belfast, UK:
     [Google Scholar]
  11. Matthews D. A., Smith W. W., Ferre R. A., Condon B., Budahazi G., Sisson W., Villafranca J. E., Janson C. A., McElroy H. E., Gribskov C. L., Worland S. 1994; Structure of human rhinovirus 3C protease reveals a trypsin-like polypeptide fold, RNA-binding Site, and means for cleaving precursor polyprotein. Cell 77:761–771
     [Google Scholar]
  12. Palmenberg A. 1990; Proteolytic processing of picornaviral polyprotein. Annual Review of Microbiology 44:603–623
     [Google Scholar]
  13. Palmenberg A. C., Parks G. D., Hall D. J., Ingraham R. H., Seng T. W., Pallai P. V. 1992; Proteolytic processing of the cardioviral P2 region: primary 2A/2B cleavage in clone-derived precursors. Virology 190:754–762
     [Google Scholar]
  14. Pevear D. C., Borkowski J., Calenoff M., Oh C. K., Ostrowski B., Lipton H. 1988; Insights into Theiler’s virus neurovirulence based upon a genomic comparison of the neurovirulent GDVII and less virulent BeAn strains. Virology 165:1–12
     [Google Scholar]
  15. Robertson B. H., Grubman M. J., Weddell G. N., Moore D. M., Welsh J. D., Fischer T., Dowbenko D. J., Yansura D. G., Small B., Kleid D. G. 1985; Nucleotide and amino acid sequence coding for polypeptides of foot-and-mouth disease virus type A12. Journal of Virology 54:651–660
     [Google Scholar]
  16. Roos R. P., Kong W.-P., Semler B. L. 1989; Polyprotein processing of Theiler’s murine encephalomyelitis virus. Journal of Virology 63:53344–5353
     [Google Scholar]
  17. 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]
  18. Ryan M. D., Belsham G. J., King A. M. Q. 1989; Specificity of substrate-enzyme interactions in foot-and-mouth disease virus polyprotein processing. Virology 173:35–45
     [Google Scholar]
  19. 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
     [Google Scholar]
  20. Sommergruber W., Zorn M., Blaas D., Fessl F., Volkmann P., Maurer-Fogy I., Pallai P., Merluzzi V., Matteo M., Skern T., Keuchler E. 1989; Polypeptide 2A of human rhinovirus type 2: identification as a protease and characterization by mutational analysis. Virology 169:68–77
     [Google Scholar]
  21. Sommergruber W., Casari G., Fessl F., Seipelt J., Skern T. 1994; The 2A proteinase of human rhinovirus is a zinc containing enzyme. Virology 204:815–818
     [Google Scholar]
  22. Toyoda H., Nicklin M. J. H., Murray M. G., Anderson C. W., Dunn J. J., Studier F. W., Wimmer E. 1986; A second virus-encoded proteinase involved in proteolytic processing of poliovirus polyprotein. Cell 45:761–770
     [Google Scholar]
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The cleavage activities of aphthovirus and cardiovirus 2A proteins.
J. Gen. Virol. 78, 13 (1997); https://doi.org/10.1099/0022-1317-78-1-13
/content/journal/jgv/10.1099/0022-1317-78-1-13
/content/journal/jgv/10.1099/0022-1317-78-1-13
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