1887

Abstract

The replicase polyproteins, pp1a and pp1ab, of porcine (TGEV) have been predicted to be cleaved by viral proteases into 16 non-structural proteins (nsp). Here, enzymic activities residing in the amino-proximal region of nsp3, the largest TGEV replicase processing product, were characterized. It was shown, by translation experiments and protein sequencing, that the papain-like protease 1, PL1, but not a mutant derivative containing a substitution of the presumed active-site nucleophile, Cys, cleaves the nsp2|nsp3 site at Gly|Gly. By using an antiserum raised against the pp1a/pp1ab residues 526–713, the upstream processing product, nsp2, was identified as an 85 kDa protein in TGEV-infected cells. Furthermore, PL1 was confirmed to be flanked at its C terminus by a domain (called X) that mediates ADP-ribose 1″-phosphatase activity. Expression and characterization of a range of bacterially expressed forms of this enzyme suggest that the active X domain comprises pp1a/pp1ab residues Asp–Ser.

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2006-03-01
2021-10-25
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