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Maturation of the Polymerase Polyprotein of the Coronavirus MHV Strain JHM Involves a Cascade of Proteolytic Processing Events

  • Jennifer J. Schiller
  • Susan C. Baker
Chapter
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Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 440)

Abstract

The RNA polymerase gene of the murine Coronavirus mouse hepatitis virus (MHV) encodes a polyprotein of greater than 750 kDa. The amino-terminal cleavage product of the MHV polymerase polyprotein, p28, has been shown to be cleaved from the polyprotein by the virus-encoded protease PCP-1. We aim to identify the MHV-JHM proteolytic products downstream of p28 and to determine which viral proteinase domains are responsible for generating each of them. To this end, we have generated antisera directed at specific MHV-JHM ORF la regions and have used these antisera to identify six viral proteins, representing a large portion of ORF la, from MHV-JHM-infected cells. These proteins include p28, p72, p65, p250, p210, and p27.

Keywords

Mouse Hepatitis Virus Murine Coronavirus Cysteine Proteinase Domain Mouse Hepatitis Virus Strain Follow Pulse Label 
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

  1. Baker, S.C., Shieh, C.K., Soe, L.H., Chang, M.F., Vannier, D.M., and Lai, M.M.C., 1989, Identification of a domain required for autoproteolytic cleavage of murine coronavirus gene A polyprotein, J. Virol. 63:3693–3699.PubMedGoogle Scholar
  2. Baker, S.C., Yokomori, K., Dong, S., Carlisle, R., Gorbalenya, A.E., Koonin, E.V., and Lai, M.M.C., 1993, Identification of the catalytic sites of a papain-like cysteine proteinase of murine coronavirus, J. Virol. 67:6056–6063.PubMedGoogle Scholar
  3. Bonilla, P.J., Hughes, S.A., and Weiss, S.R., 1997, Characterization of a second cleavage site and demonstration of activity in trans by the papain-like proteinase of the murine coronavirus mouse hepatitis virus strain A59, J. Virol. 71:900–909.PubMedGoogle Scholar
  4. Denison, M.R., Hughes, S.A., and Weiss, S.R., 1995, Identification and characterization of a 65-kDa protein processed from the gene 1 polyprotein of the murine coronavirus MHV-A59, Virology 207:316–320.PubMedCrossRefGoogle Scholar
  5. Dong, S., and Baker, S.C., 1994, Determinants of the p28 cleavage site recognized by the first papain-like cysteine proteinase of murine coronavirus, Virology 204:541–549.PubMedCrossRefGoogle Scholar
  6. Gao, H.-Q., Schiller, J.J., and Baker, S.C., 1996, Identification of the polymerase polyprotein products p72 and p65 of the murine coronavirus MHV-JHM, Virus Research 45:101–109.PubMedCrossRefGoogle Scholar
  7. Gorbalenya, A.E., Koonin, E.V., Donchenko, A.P., and Blinov, V.M., 1989, Coronavirus genome: Prediction of putative functional domains in the non-structural polyprotein by comparative amino acid sequence analysis, Nucleic Acids Res. 17:4847–4861.PubMedCrossRefGoogle Scholar
  8. Guan, K., and Dixon, J.E., 1991, Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure for fusion proteins with glutathione S-transferase, Anal. Biochem. 192:262–267.PubMedCrossRefGoogle Scholar
  9. Hughes, S.A., Bonilla, P., and Weiss, S.R., 1995, Identification of the murine coronavirus p28 cleavage site, J. Virol. 69:809–813.PubMedGoogle Scholar
  10. Lee, H.-J., Shieh, C.-K., Gorbalenya, A.E., Koonin, E.V., La Monica, N., Tuler, J., Bagdzhadzhyan, A., and Lai, M.M.C., 1991, The complete sequence (22 kilobases) of murine coronavirus gene 1 encoding the putative proteases and RNA polymerase, Virology 180:567–582.PubMedCrossRefGoogle Scholar
  11. 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.PubMedGoogle Scholar
  12. 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.PubMedCrossRefGoogle Scholar
  13. Pachuk, C.J., Bredenbeek, P.J., Zoltick, P.W, Spaan, W.J.M., and Weiss, S.R., 1989, Molecular cloning of the gene encoding the putative polymerase of mouse hepatitis coronavirus, strain A59, Virology 171:141–148.PubMedCrossRefGoogle Scholar
  14. Seybert, A., Ziebuhr, J., and Siddell, S.G., 1997, Expression and characterization of a recombinant murine coronavirus 3C-like proteinase, J. Gen. Virol. 78:71–75.PubMedGoogle Scholar
  15. 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.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1998

Authors and Affiliations

  • Jennifer J. Schiller
    • 1
  • Susan C. Baker
    • 1
  1. 1.Department of Microbiology and ImmunologyLoyola University of Chicago Stritch School of MedicineMaywoodUSA

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