title: Inducible expression of encephalomyocarditis virus 3C protease activity in stably transformed mouse cell lines.
abstract: An inducible expression vector system has been developed to facilitate the study of the effects of individual virus gene products on cell function. The vector utilizes the mouse metallothionein promoter carried on the bovine papillomavirus genome. Conditions which optimize the induced expression of open reading frames inserted downstream from the mouse metallothionein promoter have recently been described. In this communication we describe the use of this system to clone and express the encephalomyocarditis virus 3C protease in cultured mouse cells. Stably transformed cell lines could be induced to produce levels of 3C protease activity comparable to those observed during normal virus infection. In spite of this, no effects on cellular protein synthesis rate or membrane permeability were observed. It was also discovered that 3C protease as well as 3C protease-containing polyproteins are turned over. This was true not only in the induced cell clones, but also during the normal course of encephalomyocarditis virus infection, as well as in translation systems in vitro. This phenomenon was highly specific for this family of polypeptides, perhaps explaining their apparent lack of cytotoxic effects.
authors.affiliation: Department of Biology, Washington University, St. Louis, Missouri 63130-4899.
authors.forename: T G
authors.initials: TG
authors.lastname: Lawson
authors.forename: L L
authors.initials: LL
authors.lastname: Smith
authors.forename: A C
authors.initials: AC
authors.lastname: Palmenberg
authors.forename: R E
authors.initials: RE
authors.lastname: Thach
mesh_terms: D000086042:3C Viral Proteases; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002472:Cell Transformation, Viral; D003546:Cysteine Endopeptidases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D003609:Dactinomycin / Q000494:pharmacology; D004686:Encephalomyocarditis virus / Q000201:enzymology* / Q000235:genetics; D004790:Enzyme Induction; D015870:Gene Expression; D005814:Genes, Viral; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D015183:Restriction Mapping; D014764:Viral Proteins; D015678:Viral Structural Proteins / Q000235:genetics*
chemical_list: D014764:Viral Proteins; D015678:Viral Structural Proteins; D003609:Dactinomycin; D008715:Methionine; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases