title: Foot-and-mouth disease virus leader proteinase: structural insights into the mechanism of intermolecular cleavage.
abstract: Translation of foot-and-mouth disease virus RNA initiates at one of two start codons leading to the synthesis of two forms of leader proteinase L(pro) (Lab(pro) and Lb(pro)). These forms free themselves from the viral polyprotein by intra- and intermolecular self-processing and subsequently cleave the cellular eukaryotic initiation factor (eIF) 4 G. During infection, Lb(pro) removes six residues from its own C-terminus, generating sLb(pro). We present the structure of sLb(pro) bound to the inhibitor E64-R-P-NH2, illustrating how sLb(pro) can cleave between Lys/Gly and Gly/Arg pairs. In intermolecular cleavage on polyprotein substrates, Lb(pro) was unaffected by P1 or P1' substitutions and processed a substrate containing nine eIF4GI cleavage site residues whereas sLb(pro) failed to cleave the eIF4GI containing substrate and cleaved appreciably more slowly on mutated substrates. Introduction of 70 eIF4GI residues bearing the Lb(pro) binding site restored cleavage. These data imply that Lb(pro) and sLb(pro) may have different functions in infected cells.
authors.affiliation: Max F. Perutz Laboratories, Medical University of Vienna, Dr. Bohr-Gasse 9/3, A-1030 Vienna, Austria.
authors.forename: Jutta
authors.initials: J
authors.lastname: Steinberger
authors.affiliation: Max F. Perutz Laboratories, University of Vienna, Department of Structural and Computational Biology, Campus Vienna Biocenter 5, A-1030 Vienna, Austria.
authors.forename: Irina
authors.initials: I
authors.lastname: Grishkovskaya
authors.affiliation: Max F. Perutz Laboratories, Medical University of Vienna, Dr. Bohr-Gasse 9/3, A-1030 Vienna, Austria.
authors.forename: Regina
authors.initials: R
authors.lastname: Cencic
authors.affiliation: Department of Biophysics, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Três de Maio 100, 04044-20 São Paulo, Brazil.
authors.forename: Luiz
authors.initials: L
authors.lastname: Juliano
authors.affiliation: Department of Biophysics, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Três de Maio 100, 04044-20 São Paulo, Brazil.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Juliano
authors.affiliation: Max F. Perutz Laboratories, Medical University of Vienna, Dr. Bohr-Gasse 9/3, A-1030 Vienna, Austria. Electronic address: timothy.skern@meduniwien.ac.at.
authors.forename: Tim
authors.initials: T
authors.lastname: Skern
mesh_terms: D001665:Binding Sites; D002401:Cathepsin B / Q000037:antagonists & inhibitors / Q000378:metabolism; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D029544:Foot-and-Mouth Disease Virus / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D008958:Models, Molecular; D011487:Protein Conformation; D012367:RNA, Viral
chemical_list: D012367:RNA, Viral; D010450:Endopeptidases; D002401:Cathepsin B; C088747:leader proteinase, foot-and-mouth disease virus
keywords: Active site; Host cell shut-off; Initiation of protein synthesis; Papain-like cysteine proteinase; Polyprotein processing; Substrate binding