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Characterisation of a papain-like proteinase domain encoded by ORF1a of the coronavirus IBV and determination of the C-terminal cleavage site of an 87 kDa protein. 1998

K P Lim, and D X Liu
Institute of Molecular Agrobiology, Singapore.

Our previous studies have shown that two overlapping papain-like proteinase domains (PLPDs) encoded by the IBV sequence from nucleotides 4155 to 5550 is responsible for cleavage of the ORF 1a polyprotein to an 87 kDa protein. In this study, we demonstrate that only the more 5' one of the two domains, PLPD-1 encoded between nucleotides 4155 and 5031, is required for processing to the 87 kDa protein. Site-directed mutagenesis studies have shown that the Cys1274 and His1435 residues are essential for the PLPD-1 activity, suggesting that they may be the components of the catalytic centre of this proteinase. Coexpression and immunoprecipitation studies have further revealed that PLPD can interact with the 87 kDa protein. Meanwhile, data obtained from the construction and expression of a series of deletion mutants have indicated that the 87 kDa protein is encoded by the 5'-most 2600 bp part of ORF1a. further deletion and mutagenesis studies are underway to determine precisely the C-terminal cleavage site of the 87 kDa protein.

UI MeSH Term Description Entries
D010206 Papain A proteolytic enzyme obtained from Carica papaya. It is also the name used for a purified mixture of papain and CHYMOPAPAIN that is used as a topical enzymatic debriding agent. EC 3.4.22.2. Tromasin
D011498 Protein Precursors Precursors, Protein
D002384 Catalysis The facilitation of a chemical reaction by material (catalyst) that is not consumed by the reaction. Catalyses
D000086882 Coronavirus Papain-Like Proteases Papain-like proteases that occur in species of CORONAVIRIDAE. Some species have more than one papain-like protease gene. PLP-1 Protein, Coronavirus,PLP-2 Protein, Coronavirus,PLP-3 Protein, Coronavirus,Papain-Like Protease, Coronavirus,Papain-Like Proteinase 1, Coronavirus,Papain-Like Proteinase 2, Coronavirus,Papain-Like Proteinase 3, Coronavirus,Coronavirus PLP-1 Protein,Coronavirus PLP-2 Protein,Coronavirus PLP-3 Protein,Coronavirus Papain Like Proteases,PLP 1 Protein, Coronavirus,PLP 2 Protein, Coronavirus,PLP 3 Protein, Coronavirus,Papain Like Protease, Coronavirus,Papain Like Proteinase 1, Coronavirus,Papain Like Proteinase 2, Coronavirus,Papain Like Proteinase 3, Coronavirus,Papain-Like Proteases, Coronavirus
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001351 Infectious bronchitis virus A species of CORONAVIRUS causing infections in chickens and possibly pheasants. Chicks up to four weeks old are the most severely affected. Avian Infectious Bronchitis Virus,Infectious Bronchitis Virus, Avian,Infectious bronchitis viruses,bronchitis viruses, Infectious
D001665 Binding Sites The parts of a macromolecule that directly participate in its specific combination with another molecule. Combining Site,Binding Site,Combining Sites,Site, Binding,Site, Combining,Sites, Binding,Sites, Combining
D014764 Viral Proteins Proteins found in any species of virus. Gene Products, Viral,Viral Gene Products,Viral Gene Proteins,Viral Protein,Protein, Viral,Proteins, Viral
D016297 Mutagenesis, Site-Directed Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion. Mutagenesis, Oligonucleotide-Directed,Mutagenesis, Site-Specific,Oligonucleotide-Directed Mutagenesis,Site-Directed Mutagenesis,Site-Specific Mutagenesis,Mutageneses, Oligonucleotide-Directed,Mutageneses, Site-Directed,Mutageneses, Site-Specific,Mutagenesis, Oligonucleotide Directed,Mutagenesis, Site Directed,Mutagenesis, Site Specific,Oligonucleotide Directed Mutagenesis,Oligonucleotide-Directed Mutageneses,Site Directed Mutagenesis,Site Specific Mutagenesis,Site-Directed Mutageneses,Site-Specific Mutageneses

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