Functional pleiotropy of an intramolecular triplex-forming fragment from the 3′-UTR of the rat Pigr gene

A microsatellite-containing 359-bp restriction fragment, isolated from the rat Pigr gene (murine polymeric immunoglobulin receptor gene) 3′-untranslated region (3′-UTR) and inserted into 3′-UTR or 3′ flanking positions in transcription units of supercoiled plasmids, attenuates luciferase reporter gene expression in orientation- and position-dependent ways following transient transfection of human 293 cells. The same fragment stimulates orientation-dependent gene expression in a 5′ flanking position. Plasmid linearization abrogates both orientation- and position-dependent responses. Cell-free translation reveals that 5′ and 3′ flanking expression responses are proportional to increased and decreased luciferase mRNA levels, whereas 3′-UTR expression is associated with control mRNA levels. Hypersensitivity to nucleases S1 and P1, gel mobility differences between supercoiled plasmids carrying opposing microsatellite orientations, and anomalous melting profiles of this fragment are also observed. These results...