Transcriptional regulation of the human prepronociceptin gene

We have cloned regions of the 5'-untranslated region of the human prepronoc iceptin (ppN/OFQ) gene into a luciferase reporter plasmid, pGL3 Basic Vecto r. By primer extension analysis, we determined that one start site of trans cription lies within the known human cDNA sequence. There are two cyclic AM P response elements (CRE) with the consensus sequence CGTCA within 250 bp o f the start of transcription. We designed various constructs around these s ites, performed transient transfections, and measured luciferase activity i n NS20Y cells. Transcriptional activity could be regulated by a variety of factors including cAMP levels, Ca2+, and particularly by a 125-bp region ad jacent to an intron located 23 bp upstream of the translation initiating AT G. These data should help in understanding the regulatory mechanisms of ppN /OFQ gene transcription. (C) 2000 Academic Press.