Abstract
Our previous studies have shown that two overlapping papain-like proteinase domains (PLPDs) encoded by the IBV sequence from nucleotides 4155 to 5550 is responsible for cleavage of the ORF la polyprotein to an 87 kDa protein. In this study, we demonstrate that only the more 5’ one of the two domains, PLPD-1 encoded between nucleotides 4155 and 5031, is required for processing to the 87 kDa protein. Site-directed mutagenesis studies have shown that the Cys1274 and His1435 residues are essential for the PLPD-1 activity, suggesting that they may be the components of the catalytic centre of this proteinase. Coexpression and immunoprecipitation studies have further revealed that PLPD can interact with the 87 kDa protein. Meanwhile, data obtained from the construction and expression of a series of deletion mutants have indicated that the 87 kDa protein is encoded by the 5’- most 2600 bp part of ORFla. Further deletion and mutagenesis studies are underway to determine precisely the C-terminal cleavage site of the 87 kDa protein.
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Keywords
- Vero Cell
- Infectious Bronchitis Virus
- Murine Coronavirus
- Putative Catalytic Residue
- Full Length Polypeptide
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References
Boursnell, M. E. G., T. D. K. Brown, I. J. Foulds, P. F. Green, F. M. Tomley and M. M. Binns., 1987, Completion of the sequence of the genome of the Coronavirus avian infectious bronchitis virus, J. Gen. Virol. 68:57–77.
Fuerst, T. R., E. G. Niles, F. W. Studier and B. Moss., 1986, Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesises bacteriophage T7 RNA polymerase, Proc. Natl. Acad. Sci. USA 83:8122–8127.
Gorbalenya, A. E., E. Y. Koonin, A. P. Donchenko and V. M. Blinov., 1989, Coronavirus genome: prediction of putative functional domains in the non-structural polyprotein by comparative amino acid sequence analysis, Nucleic Acids Research 17:4847–4860.
Hughes, S. A., P. J. Bonilla and S. R. Weiss. 1995. Identification of the murine Coronavirus p28 cleavage site, J. Virol. 69:809–813.
Laemmli, U. K, 1970, Cleavage of structural proteins during the assembly of the bacteriophage T4. Nature (London) 227:680–685.
Lee, H-J., C-K. Shieh, A. E. Gorbalenya, E. V. Koonin, N. L. Monica, J. Tuler, A. Bagdzhadzhyan and M. M. C. Lai, 1991, The complete sequence (22 kilobases) of murine Coronavirus gene 1 encoding the putative proteases and RNA polymerase, Virology 180:567–582.
Liu, D. X., I. Brierley, K. W. Tibbies and T. D. K. Brown., 1994, A 100-kilodalton polypeptide encoded by open reading frame (ORF) lb of the Coronavirus IBVis processed by ORF la products, J. Virol. 68:5772–5780.
Liu, D. X., K. W. Tibbies, D. Cavanagh, T. D. K. Brown and I. Brierley., 1995, Identification, expression and processing of an 87 kDa polypeptide encoded by ORF1a of the Coronavirus infectious bronchitis virus, Virology 208:48–57.
Liu, D. X., H. Y. Xu, T. D. K. Brown., 1997, Proteolytic processing of the Coronavirus IBV la polyprotein: identification of a 10-kilodalton polypeptide and determination of its cleavage sites, J. Virol 208:48–57.
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© 1998 Springer Science+Business Media New York
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Lim, K.P., Liu, D.X. (1998). Characterisation of a Papain-Like Proteinase Domain Encoded by ORF1a of the Coronavirus IBV and Determination of the C-Terminal Cleavage Site of an 87 kDa Protein. In: Enjuanes, L., Siddell, S.G., Spaan, W. (eds) Coronaviruses and Arteriviruses. Advances in Experimental Medicine and Biology, vol 440. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-5331-1_22
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