Abstract
Coronavirus gene expression involves proteolytic processing of the gene 1-encoded polyproteins and a key enzyme in this process is the virus-encoded 3C-like proteinase. In this study, we describe the biosynthesis of the human Coronavirus 229E 3C-like proteinase in Escherichia coli and the substrate specificity of the purified protein. Using immunofluorescence microscopy, we have also investigated the subcellular localization of the 3C-like proteinase and have found a punctate, perinuclear distribution of the proteinase in virus-infected cells.
Chapter PDF
Similar content being viewed by others
Keywords
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
References
Grötzinger, C., Heusipp, G., Ziebuhr, J., Harms, U., Süss, J., and Siddell, S. G., 1996, Characterization of a 105-kDa polypeptide encoded in gene I of the human Coronavirus HCV 229E, Virology 222: 227—235
Herold, J. and Siddell, S. G., 1993, An elaborated pseudoknot is required for high frequency frameshifting during translation of HCV 229E polymerase mRNA, Nucleic Acids Res. 21: 5838–5842
Herold, J., Raabe, T., Schelle-Prinz, B. and Siddell, S. G., 1993, Nucleotide sequence of the human Coronavirus 229E RNA polymerase locus, Virology 195: 680–691
Heusipp, G., Grötzinger, C., Herold, J., Siddell, S. G., and Ziebuhr, J., 1997, Identification and subcellular localization of a 41-kDa, polyprotein lab processing product in human Coronavirus 229E-infected cells, J. Gen. Virol., in press
Heusipp, G., Harms, U., Siddell, S. G., and Ziebuhr, J., 1997, Identification of an ATPase activity associated withV a 71-kDa polypeptide encoded in gene 1 of the human Coronavirus 229E, J. Virol., in press
Liu, D. X. and Brown, T. D. K., 1995, Characterization and mutational analysis of an ORF la-encoding proteinase domain responsible for proteolytic processing of the infectious bronchitis virus la/ lb polyprotein, Virology 209: 420–427
Liu, D. X., Brierley, I., Tibbies., K. W., and Brown, T. D. K., 1994, A 100-kilodalton polypeptide encoded by open reading frame (ORF) lb of the Coronavirus infectious bronchitis virus is processed by ORF la products, J. Virol. 68: 5773–5780
Liu, D. X., Xu, H. Y., and Brown, T. D. K., 1997, Proteolytic processing of the Coronavirus infectious bronchitis virus la polyprotein: identification of a 10-kilodalton polypeptide and determination of its cleavage sites, J. Virol. 71 1814–1820.
Lu, Y. and Denison M. R., 1997, Determinants of mouse hepatitis virus 3C-like proteinase activity, Virology 230: 335–342.
Lu, X., Lu, Y., and Denison, M. R., 1996, Intracellular and in vitro-translated 27-kDa proteins contain the 3C-like proteinase activity of the Coronavirus MHV-A59, Virology 222 375—382.
Lu, Y., Lu, X., and Denison, M. R., 1995, Identification and characterization of a serine-like proteinase of the murine Coronavirus MHV-A59, J. Virol 69: 3554–3559.
Seybert, A., Ziebuhr, J., and Siddell, S. G., 1997, Expression and characterization of a recombinant murine Coronavirus 3C-likeproteinase,J Gen. Virol 78: 71–75.
Tibbies, K. W., Brierley, I., Cavanagh, D., and Brown, T. D. K., 1996, Characterization in vitro of an autocatalytic processing activity associated with the predicted 3C-like proteinase domain of the Coronavirus avian infectious bronchitis virus, J. Virol 70: 1923–1930.
Van Dinten, L. C., Wassenaar, A. L. M., Gorbalenya, A. E., Spaan, W. J. M., and Snijder, E. J., 1996, Processing of the equine arteritis virus replicase ORFlb protein: identification of cleavage products containing the putative viral polymerase and helicase domains, J. Virol 70: 6625–6633.
Ziebuhr, J., Herold, J., and Siddell, S. G., 1995, Characterization of a human Coronavirus (strain 229E) 3C-like proteinase activity, J. Virol 69: 4331–4338.
Ziebuhr, J., Heusipp, G., and Siddell, S. G. (1997). Biosynthesis, purification, and characterization of the human Coronavirus 229E 3C-like proteinase, J. Virol 71: 3992–3997.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1998 Springer Science+Business Media New York
About this chapter
Cite this chapter
Ziebuhr, J., Heusipp, G., Seybert, A., Siddell, S.G. (1998). Substrate Specificity of the Human Coronavirus 229E 3C-Like Proteinase. 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_15
Download citation
DOI: https://doi.org/10.1007/978-1-4615-5331-1_15
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-7432-9
Online ISBN: 978-1-4615-5331-1
eBook Packages: Springer Book Archive