title: Pre-mRNA processing enhancer (PPE) element increases the expression of an intronless thymidylate synthase gene but does not affect intron-dependent S phase regulation.
abstract: The pre-mRNA processing enhancer (PPE) element is an RNA sequence element derived from the intronless HSV-TK gene. Insertion of the element into the highly intron-dependent human beta-globin gene leads to efficient expression in the absence of splicing. We have analyzed the effect of the PPE element on the expression of mouse thymidylate synthase (TS) minigenes. We have previously shown that the expression of intronless TS minigenes is moderately (up to 20-fold) stimulated by the inclusion of introns. Furthermore, S phase-specific expression of TS minigenes in growth-stimulated cells depends on the presence of a spliceable intron as well as the TS promoter. The goal of our study was to determine if the PPE element would overcome the dependence on introns for efficient expression and for S phase-specific expression of transfected TS minigenes. We found that insertion of the PPE element into an intronless TS minigene partially overcame intron dependence. However, the increase in expression was much less than that observed for the intronless beta-globin gene. We also found that intronless TS or HSV-TK genes that contained the PPE element and that were driven by the TS promoter were expressed at a constant level in serum-stimulated cells. However, when an intron was included in these genes, they were expressed in an S phase-specific manner. Thus the PPE element was not able to overcome the dependence on introns for S phase-specific expression of TS minigenes.
authors.affiliation: Department of Molecular Genetics, The Ohio State University, Columbus 43210, USA.
authors.forename: T X
authors.initials: TX
authors.lastname: Lee
authors.forename: L F
authors.initials: LF
authors.lastname: Johnson
mesh_terms: D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002455:Cell Division / Q000235:genetics; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004742:Enhancer Elements, Genetic / Q000502:physiology*; D005786:Gene Expression Regulation; D007438:Introns / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D012322:RNA Precursors / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D016196:S Phase / Q000235:genetics*; D013937:Thymidine Kinase / Q000235:genetics; D013940:Thymidylate Synthase / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
chemical_list: D012322:RNA Precursors; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D013940:Thymidylate Synthase; D013937:Thymidine Kinase