Analysis of the stimulation of reporter gene expression by the sigma 3 protein of reovirus in co-transfected cells.
The stimulation of reporter gene expression following co-transfection with the S4 gene of mammalian reoviruses was analysed. The sigma 3 protein of type 3 reovirus gave a five- to eightfold increase in expression of chloramphenicol acetyltransferase and beta-galactosidase but this was found to be dependent upon the nature of the promoter being used to drive reporter gene expression. The sigma 3 protein of reovirus type 1 failed to stimulate reporter gene expression under any of the conditions used. Hybrid constructs between the S4 genes of reoviruses type 1 and 3 were used to map the stimulation characteristic to the carboxy-terminal third of the gene. Analysis of the level of sigma 3 protein accumulation in transfected cells showed that the reovirus type 3 protein accumulated to a much higher level than that of reovirus type 1. Using the hybrid gene constructs this higher level of protein accumulation was shown to co-segregate with the ability to stimulate reporter gene expression.
Department of Biological Sciences, University of Warwick, Coventry, U.K.
P E
PE
Martin
M A
MA
McCrae
D000818:Animals; D036022:Capsid Proteins; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000235:genetics; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral / Q000235:genetics*; D012089:Mammalian orthoreovirus 3 / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014162:Transfection; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D036022:Capsid Proteins; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C053179:sigma protein 3, Reovirus; D015500:Chloramphenicol O-Acetyltransferase; D001616:beta-Galactosidase