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 polymerase chain reaction. This 1.7 kb region contains several potential binding sites for transcription factors, among which are binding sites for TF-IID, cyclic AMP response element binding protein, glucocorticoid receptor and estrogen receptor. Multiple start points for the transcription of prepro-nociceptin are identified by an 'oligoribonucleotide-capping' method, but the major one is located at -558(G). Promoter activity assays using luciferase reporter 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 cellular cyclic AMP levels by forskolin treatment up-regulates prepro-nociceptin transcription. Estrogen also up-regulates gene transcription whereas glucocorticoid down-regulates transcription, each through their corresponding receptor response elements. These regulatory effects can be blocked either by mutations of the potential cyclic AMP- or estrogen receptor response elements or by the application of antagonists for glucocorticoid and estrogen receptors. These findings provide a molecular basis for the regulatory mechanisms of human prepro-nociceptin gene expression.
Xie
G X
GX
Department of Anesthesia, University of California, San Francisco 94143, USA.
Ito
E
E
Maruyama
K
K
Suzuki
Y
Y
Sugano
S
S
Sharma
M
M
Pietruck
C
C
Palmer
P P
PP
D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000494:pharmacology*; D004247:DNA; D004967:Estrogens / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000502:physiology; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011498:Protein Precursors / Q000235:genetics*; D011960:Receptors, Estrogen / Q000502:physiology; D011965:Receptors, Glucocorticoid / Q000502:physiology; D011957:Receptors, Opioid / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism
D004967:Estrogens; D005938:Glucocorticoids; D011498:Protein Precursors; D011960:Receptors, Estrogen; D011965:Receptors, Glucocorticoid; D011957:Receptors, Opioid; D014157:Transcription Factors; C121527:prepronociceptin; D004247:DNA; D000242:Cyclic AMP