title: SARS hCoV papain-like protease is a unique Lys48 linkage-specific di-distributive deubiquitinating enzyme.
abstract: Ubiquitin (Ub) and the Ub-like (Ubl) modifier interferon-stimulated gene 15 (ISG15) participate in the host defence of viral infections. Viruses, including the severe acute respiratory syndrome human coronavirus (SARS hCoV), have co-opted Ub-ISG15 conjugation pathways for their own advantage or have evolved effector proteins to counter pro-inflammatory properties of Ub-ISG15-conjugated host proteins. In the present study, we compare substrate specificities of the papain-like protease (PLpro) from the recently emerged Middle East respiratory syndrome (MERS) hCoV to the related protease from SARS, SARS PLpro. Through biochemical assays, we show that, similar to SARS PLpro, MERS PLpro is both a deubiquitinating (DUB) and a deISGylating enzyme. Further analysis of the intrinsic DUB activity of these viral proteases revealed unique differences between the recognition and cleavage specificities of polyUb chains. First, MERS PLpro shows broad linkage specificity for the cleavage of polyUb chains, whereas SARS PLpro prefers to cleave Lys48-linked polyUb chains. Secondly, MERS PLpro cleaves polyUb chains in a 'mono-distributive' manner (one Ub at a time) and SARS PLpro prefers to cleave Lys48-linked polyUb chains by sensing a di-Ub moiety as a minimal recognition element using a 'di-distributive' cleavage mechanism. The di-distributive cleavage mechanism for SARS PLpro appears to be uncommon among USP (Ub-specific protease)-family DUBs, as related USP family members from humans do not display such a mechanism. We propose that these intrinsic enzymatic differences between SARS and MERS PLpro will help to identify pro-inflammatory substrates of these viral DUBs and can guide in the design of therapeutics to combat infection by coronaviruses.
authors.affiliation: *Department of Biochemistry & Molecular Pharmacology, New York University School of Medicine, New York, NY 10016, U.S.A.
authors.forename: Miklós
authors.initials: M
authors.lastname: Békés
authors.affiliation: †Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology, Wyb. Wyspianskiego 27, Wroclaw 50-370, Poland.
authors.forename: Wioletta
authors.initials: W
authors.lastname: Rut
authors.affiliation: †Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology, Wyb. Wyspianskiego 27, Wroclaw 50-370, Poland.
authors.forename: Paulina
authors.initials: P
authors.lastname: Kasperkiewicz
authors.affiliation: ‡Division of Cell Biology, Netherlands Cancer Institute (NKI), Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands.
authors.forename: Monique P C
authors.initials: MP
authors.lastname: Mulder
authors.affiliation: ‡Division of Cell Biology, Netherlands Cancer Institute (NKI), Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands.
authors.forename: Huib
authors.initials: H
authors.lastname: Ovaa
authors.affiliation: †Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology, Wyb. Wyspianskiego 27, Wroclaw 50-370, Poland.
authors.forename: Marcin
authors.initials: M
authors.lastname: Drag
authors.affiliation: §Structural Biology Program, Sloan-Kettering Institute, 1275 York Ave, New York, NY 10065, U.S.A.
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Lima
authors.affiliation: *Department of Biochemistry & Molecular Pharmacology, New York University School of Medicine, New York, NY 10016, U.S.A.
authors.forename: Tony T
authors.initials: TT
authors.lastname: Huang
mesh_terms: D018352:Coronavirus Infections / Q000378:metabolism / Q000821:virology; D010450:Endopeptidases / Q000378:metabolism*; D006801:Humans; D008239:Lysine / Q000378:metabolism*; D010206:Papain / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism*; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D045473:Severe acute respiratory syndrome-related coronavirus / Q000201:enzymology*; D013379:Substrate Specificity; D025801:Ubiquitin / Q000378:metabolism*; D054875:Ubiquitination; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D025801:Ubiquitin; D014764:Viral Proteins; D010450:Endopeptidases; D010447:Peptide Hydrolases; D010206:Papain; D008239:Lysine