1887

Journal of General Virology header logo

Volume 96, Issue 9

Cleavage of bovine adenovirus type 3 non-structural 100K protein by protease is required for nuclear localization in infected cells but is not essential for virus replication Free

Abstract

The L6 region of bovine adenovirus type 3 (BAdV-3) encodes a non-structural protein named 100K. Rabbit antiserum raised against BAdV-3 100K recognized a protein of 130 kDa at 12–24 h and proteins of 130, 100, 95 and 15 kDa at 36–48 h after BAdV-3 infection. The 100K species localized to the nucleus and the cytoplasm of BAdV-3-infected cells. In contrast, 100K localized predominantly to the cytoplasm of the transfected cells. However, BAdV-3 infection of cells transfected with 100K–enhanced yellow fluorescent protein-expressing plasmid detected fluorescent protein in the nucleus of the cells, suggesting that other viral proteins may be required for the nuclear localization of 100K. Interaction of BAdV-3 100K with BAdV-3 33K protein did not alter the cytoplasmic localization of 100K. However, co-expression of BAdV-3 100K and BAdV-3 protease localized 100K to the nucleolus of the transfected cells. Subsequent analysis suggested that BAdV-3 protease cleaves 100K at two identified potential protease cleavage sites (aa 740–745 and 781–786) in transfected or BAdV-3-infected cells. The cleaved C terminus (107 aa) was localized to the nucleolus of the transfected cells. Further analysis suggested that the cleaved C terminus contains a bipartite nuclear localization signal and utilizes import receptor importin-α3 of the classical importin-α/β transport pathway for nuclear transport. Successful isolation of recombinant BAdV-3 expressing mutant 100K (substitution of alanine for glycine in the potential protease cleavage site) suggested that cytoplasmic cleavage of BAdV-3 100K by adenoviral protease is not essential for virus replication.

  • Received:
  • Accepted:
  • Published Online:
© 2015 The Authors
Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.000205
2015-09-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/96/9/2749.html?itemId=/content/journal/jgv/10.1099/vir.0.000205&mimeType=html&fmt=ahah

References

  1. Andrade F., Bull H.G., Thornberry N.A., Ketner G.W., Casciola-Rosen L.A., Rosen A. 2001; Adenovirus L4-100K assembly protein is a granzyme B substrate that potently inhibits granzyme B-mediated cell death. Immunity 14:751–761 [View Article][PubMed]
     [Google Scholar]
  2. Ayalew L.E. 2014 The role of bovine adenovirus (BAdV-3) protein pVIII in virus replication PhD thesis University of Saskatchewan: Saskatoo, Canada;
     [Google Scholar]
  3. Berk A.J. 2007; Adenoviridae: the viruses and their replication. In Fields Virology, 5th edn. pp. 2355–2394 Edited by Knipe P. H. D. Philadelphia: Wolters Kluwer Health/Lippincott Williams & Wilkins;
     [Google Scholar]
  4. Blanchette P., Wimmer P., Dallaire F., Cheng C.Y., Branton P.E. 2013; Aggresome formation by the adenoviral protein E1B55K is not conserved among adenovirus species and is not required for efficient degradation of nuclear substrates. J Virol 87:4872–4881 [View Article][PubMed]
     [Google Scholar]
  5. Chambers T.J., Weir R.C., Grakoui A., McCourt D.W., Bazan J.F., Fletterick R.J., Rice C.M. 1990; Evidence that the N-terminal domain of nonstructural protein NS3 from yellow fever virus is a serine protease responsible for site-specific cleavages in the viral polyprotein. Proc Natl Acad Sci U S A 87:8898–8902 [View Article][PubMed]
     [Google Scholar]
  6. Cheng C.Y., Gilson T., Wimmer P., Schreiner S., Ketner G., Dobner T., Branton P.E., Blanchette P. 2013; Role of E1B55K in E4orf6/E1B55K E3 ligase complexes formed by different human adenovirus serotypes. J Virol 87:6232–6245 [View Article][PubMed]
     [Google Scholar]
  7. Cuesta R., Xi Q., Schneider R.J. 2000; Adenovirus-specific translation by displacement of kinase Mnk1 from cap-initiation complex eIF4F. EMBO J 19:3465–3474 [View Article][PubMed]
     [Google Scholar]
  8. Diouri M., Keyvani-Amineh H., Geoghegan K.F., Weber J.M. 1996; Cleavage efficiency by adenovirus protease is site-dependent. J Biol Chem 271:32511–32514 [View Article][PubMed]
     [Google Scholar]
  9. Du E., Tikoo S.K. 2010; Efficient replication and generation of recombinant bovine adenovirus-3 in nonbovine cotton rat lung cells expressing I-SceI endonuclease. J Gene Med 12:840–847 [View Article][PubMed]
     [Google Scholar]
  10. Gao M., Matusick-Kumar L., Hurlburt W., DiTusa S.F., Newcomb W.W., Brown J.C., McCann P.J. III, Deckman I., Colonno R.J. 1994; The protease of herpes simplex virus type 1 is essential for functional capsid formation and viral growth. J Virol 68:3702–3712[PubMed]
     [Google Scholar]
  11. Greber U.F., Webster P., Weber J., Helenius A. 1996; The role of the adenovirus protease on virus entry into cells. EMBO J 15:1766–1777[PubMed]
     [Google Scholar]
  12. Hong S.S., Szolajska E., Schoehn G., Franqueville L., Myhre S., Lindholm L., Ruigrok R.W.H., Boulanger P., Chroboczek J. 2005; The 100K-chaperone protein from adenovirus serotype 2 (Subgroup C) assists in trimerization and nuclear localization of hexons from subgroups C and B adenoviruses. J Mol Biol 352:125–138 [View Article][PubMed]
     [Google Scholar]
  13. Kohl N.E., Emini E.A., Schleif W.A., Davis L.J., Heimbach J.C., Dixon R.A., Scolnick E.M., Sigal I.S. 1988; Active human immunodeficiency virus protease is required for viral infectivity. Proc Natl Acad Sci U S A 85:4686–4690 [View Article][PubMed]
     [Google Scholar]
  14. Köhler M., Görlich D., Hartmann E., Franke J. 2001; Adenoviral E1A protein nuclear import is preferentially mediated by importin alpha3 in vitro . Virology 289:186–191 [View Article][PubMed]
     [Google Scholar]
  15. Koyuncu O.O., Dobner T. 2009; Arginine methylation of human adenovirus type 5 L4 100-kilodalton protein is required for efficient virus production. J Virol 83:4778–4790 [View Article][PubMed]
     [Google Scholar]
  16. Kulshreshtha V., Tikoo S.K. 2008; Interaction of bovine adenovirus-3 33K protein with other viral proteins. Virology 381:29–35 [View Article][PubMed]
     [Google Scholar]
  17. Kulshreshtha V., Babiuk L.A., Tikoo S.K. 2004; Role of bovine adenovirus-3 33K protein in viral replication. Virology 323:59–69 [View Article][PubMed]
     [Google Scholar]
  18. Mangel W.F., McGrath W.J., Toledo D.L., Anderson C.W. 1993; Viral DNA and a viral peptide can act as cofactors of adenovirus virion proteinase activity. Nature 361:274–275 [View Article][PubMed]
     [Google Scholar]
  19. Mangel W.F., Baniecki M.L., McGrath W.J. 2003; Specific interactions of the adenovirus proteinase with the viral DNA, an 11-amino-acid viral peptide, and the cellular protein actin. Cell Mol Life Sci 60:2347–2355 [View Article][PubMed]
     [Google Scholar]
  20. Patel A.K., Olson D., Tikoo S.K. 2010; Proteomic analysis of bovine nucleolus. Genomics Proteomics Bioinformatics 8:145–158 [View Article][PubMed]
     [Google Scholar]
  21. Paterson C.P. 2010 Molecular characterization of 52K protein of bovine adenovirus type 3 PhD thesis University of Saskatchewan: Saskatoon, Canada;
     [Google Scholar]
  22. Paterson C.P., Ayalew L.E., Tikoo S.K. 2012; Mapping of nuclear import signal and importin α3 binding regions of 52K protein of bovine adenovirus-3. Virology 432:63–72 [View Article][PubMed]
     [Google Scholar]
  23. Pérez-Berná A.J., Mangel W.F., McGrath W.J., Graziano V., Flint J., San Martín C. 2014; Processing of the l1 52/55k protein by the adenovirus protease: a new substrate and new insights into virion maturation. J Virol 88:1513–1524 [View Article][PubMed]
     [Google Scholar]
  24. Reddy P.S., Idamakanti N., Song J.Y., Lee J.B., Hyun B.H., Park J.H., Cha S.H., Bae Y.T., Tikoo S.K., Babiuk L.A. 1998; Nucleotide sequence and transcription map of porcine adenovirus type 3. Virology 251:414–426 [View Article][PubMed]
     [Google Scholar]
  25. Ruzindana-Umunyana A., Sircar S., Weber J.M. 2000; The effect of mutant peptide cofactors on adenovirus protease activity and virus infection. Virology 270:173–179 [View Article][PubMed]
     [Google Scholar]
  26. Ruzindana-Umunyana A., Imbeault L., Weber J.M. 2002; Substrate specificity of adenovirus protease. Virus Res 89:41–52 [View Article][PubMed]
     [Google Scholar]
  27. Stracker T.H., Lee D.V., Carson C.T., Araujo F.D., Ornelles D.A., Weitzman M.D. 2005; Serotype-specific reorganization of the Mre11 complex by adenoviral E4orf3 proteins. J Virol 79:6664–6673 [View Article][PubMed]
     [Google Scholar]
  28. Weber J. 1976; Genetic analysis of adenovirus type 2 III. Temperature sensitivity of processing viral proteins. J Virol 17:462–471[PubMed]
     [Google Scholar]
  29. Weber J. 1995; Adenovirus endopeptidase and its role in virus infection. In The Molecular Repertoire of Adenoviruses I pp. 227–235 Edited by Doerfler W., Böhm P. Berlin/Heidelberg: Springer-Verlag; [CrossRef]
     [Google Scholar]
  30. Wodrich H., Guan T., Cingolani G., Von Seggern D., Nemerow G., Gerace L. 2003; Switch from capsid protein import to adenovirus assembly by cleavage of nuclear transport signals. EMBO J 22:6245–6255 [View Article][PubMed]
     [Google Scholar]
  31. Wu Q., Chen Y., Kulshreshtha V., Tikoo S.K. 2004; Characterization and nuclear localization of the fiber protein encoded by the late region 7 of bovine adenovirus type 3. Arch Virol 149:1783–1799 [View Article][PubMed]
     [Google Scholar]
  32. Xi Q., Cuesta R., Schneider R.J. 2004; Tethering of eIF4G to adenoviral mRNAs by viral 100k protein drives ribosome shunting. Genes Dev 18:1997–2009 [View Article][PubMed]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.000205
Loading
Cleavage of bovine adenovirus type 3 non-structural 100K protein by protease is required for nuclear localization in infected cells but is not essential for virus replication
J. Gen. Virol. 96, 2749 (2015); https://doi.org/10.1099/vir.0.000205
/content/journal/jgv/10.1099/vir.0.000205
/content/journal/jgv/10.1099/vir.0.000205
Loading

Data & Media loading...

Supplements

Supplementary Data

PDF

Most cited 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