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. 2010 Jan;1(1):59-74.
doi: 10.1007/s13238-010-0011-4. Epub 2010 Feb 7.

Liberation of SARS-CoV main protease from the viral polyprotein: N-terminal autocleavage does not depend on the mature dimerization mode

Shuai Chen  1 , Felix Jonas, Can Shen, Rolf Hilgenfeld
Affiliations

Liberation of SARS-CoV main protease from the viral polyprotein: N-terminal autocleavage does not depend on the mature dimerization mode

Shuai Chen et al. Protein Cell. 2010 Jan.

Erratum in

  • Protein Cell. 2010 Mar;1(3):307. Higenfeld, Rolf [corrected to Hilgenfeld, Rolf]

Abstract

The main protease (M(pro)) plays a vital role in proteolytic processing of the polyproteins in the replicative cycle of SARS coronavirus (SARS-CoV). Dimerization of this enzyme has been shown to be indispensable for trans-cleavage activity. However, the auto-processing mechanism of M(pro), i.e. its own release from the polyproteins through autocleavage, remains unclear. This study elucidates the relationship between the N-terminal autocleavage activity and the dimerization of "immature" M(pro). Three residues (Arg4, Glu290, and Arg298), which contribute to the active dimer conformation of mature M(pro), are selected for mutational analyses. Surprisingly, all three mutants still perform N-terminal autocleavage, while the dimerization of mature protease and trans-cleavage activity following auto-processing are completely inhibited by the E290R and R298E mutations and partially so by the R4E mutation. Furthermore, the mature E290R mutant can resume N-terminal autocleavage activity when mixed with the "immature" C145A/E290R double mutant whereas its trans-cleavage activity remains absent. Therefore, the N-terminal auto-processing of M(pro) appears to require only two "immature" monomers approaching one another to form an "intermediate" dimer structure and does not strictly depend on the active dimer conformation existing in mature protease. In conclusion, an auto-release model of M(pro) from the polyproteins is proposed, which will help understand the auto-processing mechanism and the difference between the autocleavage and trans-cleavage proteolytic activities of SARS-CoV M(pro).

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References

    1. Anand, K., Yang, H., Bartlam, M., Rao, Z., and Hilgenfeld, R. (2005). Coronavirus main proteinase: target for antiviral drug therapy. In Coronaviruses with special emphasis on first insights concerning SARS. A. Schmidt, M.H. Wolf and O. Weber, ed. (Basel, Birkhäuser) pp. 173–199.
    1. Anand K., Ziebuhr J., Wadhwani P., Mesters J.R., Hilgenfeld R. Coronavirus main proteinase (3CLpro) structure: basis for design of anti-SARS drugs. Science. 2003;300:1763–1767. doi: 10.1126/science.1085658. - DOI - PubMed
    1. Barrila J., Bacha U., Freire E. Long-range cooperative interactions modulate dimerization in SARS 3CLpro. Biochemistry. 2006;45:14908–14916. doi: 10.1021/bi0616302. - DOI - PMC - PubMed
    1. Chen S., Chen L., Tan J., Chen J., Du L., Sun T., Shen J., Chen K., Jiang H., Shen X. Severe acute respiratory syndrome coronavirus 3C-like proteinase N terminus is indispensable for proteolytic activity but not for enzyme dimerization. Biochemical and thermodynamic investigation in conjunction with molecular dynamics simulations. J Biol Chem. 2005;280:164–173. doi: 10.1074/jbc.M408211200. - DOI - PMC - PubMed
    1. Chen S., Hu T., Zhang J., Chen J., Chen K., Ding J., Jiang H., Shen X. Mutation of Gly-11 on the dimer interface results in the complete crystallographic dimer dissociation of severe acute respiratory syndrome coronavirus 3C-like protease: crystal structure with molecular dynamics simulations. J Biol Chem. 2008;283:554–564. doi: 10.1074/jbc.M705240200. - DOI - PMC - PubMed

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