Transcriptional regulation of the human prepronociceptin gene.
We have cloned regions of the 5'-untranslated region of the human prepronociceptin (ppN/OFQ) gene into a luciferase reporter plasmid, pGL3 Basic Vector. By primer extension analysis, we determined that one start site of transcription lies within the known human cDNA sequence. There are two cyclic AMP response elements (CRE) with the consensus sequence CGTCA within 250 bp of the start of transcription. We designed various constructs around these sites, performed transient transfections, and measured luciferase activity in NS20Y cells. Transcriptional activity could be regulated by a variety of factors including cAMP levels, Ca(2+), and particularly by a 125-bp region adjacent to an intron located 23 bp upstream of the translation initiating ATG. These data should help in understanding the regulatory mechanisms of ppN/OFQ gene transcription.
Zaveri
N T
NT
Pharmaceutical Discovery Division, SRI International, 333 Ravenswood Avenue, Menlo Park, California 94025, USA. nina@topaz.sri.com
Green
C J
CJ
Toll
L
L
D020121:5' Untranslated Regions / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000032:analysis; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000235:genetics*; D011957:Receptors, Opioid / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic
D020121:5' Untranslated Regions; D018076:DNA, Complementary; D011498:Protein Precursors; D011957:Receptors, Opioid; D014157:Transcription Factors; C121527:prepronociceptin; D008156:Luciferases