The promoter region of human prepro-nociceptin gene and its regulation by cyclic AMP and steroid hormones

The promoter and upstream regulatory region of the human prepro-nociceptin gene has been cloned from adaptor-ligated genomic DNA libraries by polymera se chain reaction. This 1.7 kb region contains several potential binding si tes for transcription factors, among which are binding sites for TF-IID, cy clic AMP response element binding protein, glucocorticoid receptor and estr ogen receptor. Multiple start points for the transcription of prepro-nocice ptin are identified by an 'oligoribonucleotide-capping' method, but the maj or one is located at -558(G). Promoter activity assays using luciferase rep orter gene constructions with the 1.7 kb fragment and a series of deletion mutations demonstrate that the core promoter is located in the region from -639 to -521 (a region surrounding the major transcription start point -558 ). A TATA-box motif displays weak promoter activity. An increase of cellula r cyclic AMP levels by forskolin treatment up-regulates prepro-nociceptin t ranscription. Estrogen also up-regulates gene transcription whereas glucoco rticoid down-regulates transcription, each through their corresponding rece ptor response elements. These regulatory effects can be blocked either by m utations of the potential cyclic AMP- or estrogen receptor-response element s or by the application of antagonists for glucocorticoid and estrogen rece ptors. These findings provide a molecular basis for the regulatory mechanis ms of human prepro-nociceptin gene expression. (C) 1999 Elsevier Science B. V. All rights reserved.