title: Post-transcriptional regulation in higher eukaryotes: the role of the reporter gene in controlling expression.
abstract: We have investigated whether reporter genes influence cytoplasmic regulation of gene expression in tobacco and Chinese hamster ovary (CHO) cells. Two genes, uidA encoding beta-glucuronidase (GUS) from Escherichia coli and Luc, encoding firefly luciferase (LUC), were used to analyze the ability of a cap, polyadenylated tail, and the 5'- and 3'-untranslated regions (UTR) from tobacco mosaic virus (TMV) to regulate expression. The regulation associated with the 5' cap structure and the TMV 5'-UTR, both of which enhance translational efficiency, was reporter gene-independent. The poly(A) tail and the TMV 3'-UTR, which is functionally equivalent to a poly(A) tail, increase translational efficiency as well as mRNA stability. The regulation associated with these 3' ends was highly reporter gene-dependent; their effect on GUS expression was almost an order of magnitude greater than that on LUC expression. In tobacco, the tenfold reporter gene effect on poly(A) tail or TMV 3'-UTR function could not be explained by a differential impact on mRNA stability; GUS and LUC mRNA half-life increased only twofold when either the poly(A) tail or TMV 3'-UTR was present. In CHO cells, however, GUS mRNA was stabilized to a greater extent by a poly(A) tail or the TMV 3'-UTR than was LUC mRNA.
authors.affiliation: Department of Biological Sciences, Stanford University, CA 94305-5020.
authors.forename: D R
authors.initials: DR
authors.lastname: Gallie
authors.forename: J N
authors.initials: JN
authors.lastname: Feder
authors.forename: R T
authors.initials: RT
authors.lastname: Schimke
authors.forename: V
authors.initials: V
authors.lastname: Walbot
mesh_terms: D000818:Animals; D001483:Base Sequence; D006224:Cricetinae; D003593:Cytoplasm / Q000502:physiology; D005819:Genetic Markers; D005966:Glucuronidase / Q000096:biosynthesis; D006207:Half-Life; D008156:Luciferases / Q000096:biosynthesis; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D010957:Plasmids; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000378:metabolism; D012808:Sigma Factor / Q000502:physiology; D014026:Nicotiana; D014027:Tobacco Mosaic Virus
chemical_list: D005819:Genetic Markers; D012333:RNA, Messenger; D012808:Sigma Factor; D008156:Luciferases; D005966:Glucuronidase