1887

Abstract

SUMMARY

Peptides were synthesized based on the cleavage sites in the adenovirus type 2 proteins pVI and pVII. The synthetic peptides were incubated with disrupted, purified adenovirus as a source of proteinase and specific cleavages were monitored by fast protein liquid chromatography and amino acid analysis. Using this approach it was established that all the peptides cleaved were of the form M(L)XGX↓G or M(L)XGG↓X. Thus we have shown that the adenoviral proteinase recognizes a specific secondary structure formed by a sequence of at least five amino acids, the main determinants of specificity being two and four residues to the N-terminal side of the bond cleaved. We were able to examine the relevant structural features of the peptide substrates by utilizing the CHEM-X molecular modelling package. Using our consensus sequence we were able to predict the cleavage sites in the viral proteins pVIII, pre-terminal protein (pTP), 11K and IIIa. Octapeptides containing the predicted sites in pVIII and the pTP were synthesized and shown to be cleaved by the proteinase.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-70-12-3225
1989-12-01
2022-01-23
Loading full text...

Full text loading...

/deliver/fulltext/jgv/70/12/JV0700123225.html?itemId=/content/journal/jgv/10.1099/0022-1317-70-12-3225&mimeType=html&fmt=ahah

References

  1. Anderson C. W., Baum P. R., Gesteland R. F. 1973; Processing of adenovirus 2-induced proteins. Journal of Virology 12:241–252
    [Google Scholar]
  2. Akusjarvi G., Persson H. 1981; Gene and mRNA for precursor polypeptide VI from adenovirus type 2. Journal of Virology 38:469–482
    [Google Scholar]
  3. Berger A., Schechter I. 1970; Mapping the active site of papain with the aid of peptide substrates and inhibitors. Philosophical Transactions of the Royal Society of London, Series B 257:249–264
    [Google Scholar]
  4. Bhatti A. R., Weber J. 1979; Protease of adenovirus type 2: partial characterisation. Virology 96:478–485
    [Google Scholar]
  5. Boudin M.-L., D’Halluin J.-C., Cousin C., Boulanger P. 1980; Human adenovirus type 2 protein Ilia: II. Maturation and encapsidation. Virology 101:144–156
    [Google Scholar]
  6. Hannan C., Raptis L. H., Dery C. V., Weber J. 1983; Biological and structural studies with an adenovirus type 2 temperature-sensitive mutant defective for uncoating. Intervirology 19:213–223
    [Google Scholar]
  7. Krausslich H., Wimmer E. 1988; Viral proteinases. Annual Review of Biochemistry 57:701–754
    [Google Scholar]
  8. Leith I. R., Hay R. T., Russell W. C. 1989; Adenovirus subviral particles and cores can support limited DNA replication. Journal of General Virology 70:3235–3248
    [Google Scholar]
  9. Miles B. D., Luftig R. B., Weatherbee J. A., Weihing R. R., Weber J. 1980; Quantitation of the interaction between adenovirus types 2 and 5 and microtubules inside infected cells. Virology 105:265–269
    [Google Scholar]
  10. Polgar L. 1987; Structure and function of serine proteases. In Hydrolytic Enzymes159–200 Neuberger A., Brocklehurst K. Amsterdam: Elsevier;
    [Google Scholar]
  11. Roberts M. M., White J. L., Grutter M. G., Burnett R. M. 1986; Three-dimensional structure of the adenovirus major coat protein hexon. Science 232:1148–1151
    [Google Scholar]
  12. Russell W. C., Blair G. E. 1977; Polypeptide phosphorylation in adenovirus-infected cells. Journal of General Virology 34:19–35
    [Google Scholar]
  13. Smart J. E., Stillman B. W. 1982; Adenovirus terminal protein precursor: partial amino acid sequence and the site of covalent linkage to DNA. Journal of Biological Chemistry 257:13499–13506
    [Google Scholar]
  14. Sung M. T., Cao T. M., Lischwe M. A., Coleman R. T. 1983; Molecular processing of adenovirus proteins. Journal of Biological Chemistry 258:8266–8272
    [Google Scholar]
  15. Staden R. 1986; The current status and portability of our sequence handling software. Nucleic Acids Research 114:217–231
    [Google Scholar]
  16. Toogood C. I. A., Murali R., Burnett R. M., Hay R. T. 1989; The adenovirus type 40 hexon: sequence, predicted structure and relationship to other adenovirus hexons. Journal of General Virology 70:3203–3248
    [Google Scholar]
  17. Tremblay M. L., Dery C. V., Talbot B. G., Weber J. 1983; In vitro cleavage specificity of the adenovirus type 2 proteinase. Biochimica et biophysica acta 743:239–245
    [Google Scholar]
  18. Weber J. 1976; Genetic analysis of adenovirus type 2: III. Temperature sensitivity of processing of viral proteins. Journal of Virology 17:462–471
    [Google Scholar]
  19. Weber J., Anderson C. W. 1988; Identification of the gene coding for the precursor of adenovirus core protein X. Journal of Virology 62:1741–1745
    [Google Scholar]
  20. Webster A., Russell W. C., Kemp G. D. 1989; Characterization of the adenovirus proteinase; development and use of a specific peptide assay. Journal of General Virology 70:3215–3223
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-70-12-3225
Loading
/content/journal/jgv/10.1099/0022-1317-70-12-3225
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