c-Jun activation of the DYNCRE3 site in the prodynorphin promoter.
The DYNCRE3 site in the prodynorphin promoter is similar to both the AP-1 and cAMP-responsive element (CRE) consensus sequences. Because c-Jun is known to bind to both AP-1 and CRE sequences, we evaluated the potential role of this transcription factor at the DYNCRE3 site using transient transfection and gel mobility shift analyses. In PC12 cells, co-transfections of a chloramphenicol acetyl transferase (CAT) reporter gene containing the DYNCRE3 site and a c-Jun expression vector resulted in transcriptional activity 9-fold greater than control. Co-transfections with a mutant c-Jun protein lacking the transactivation domain resulted in a concentration-dependent decrease in transcriptional activity. Gel mobility shift analysis demonstrated the formation of a multi-component protein-DNA complex between an oligonucleotide centered on the DYNCRE3 site and nuclear extract from untreated and forskolin-stimulated PC12 cells. The upper band of this complex could be completely supershifted with the addition of a c-Jun specific antibody. These convergent data suggest that c-Jun is involved in transcriptional activation through the DYNCRE3 site.
Messersmith
D J
DJ
Neurobiology and Anesthesiology Branch, National Institute of Dental Research, National Institutes of Health, Bethesda, MD 20892, USA.
Kim
D J
DJ
Gu
J
J
Dubner
R
R
Iadarola
M J
MJ
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D005576:Colforsin / Q000494:pharmacology; D016384:Consensus Sequence; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D004745:Enkephalins / Q000096:biosynthesis / Q000235:genetics*; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016716:PC12 Cells; D011401:Promoter Regions, Genetic; D011498:Protein Precursors / Q000096:biosynthesis / Q000235:genetics*; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection
D017362:Cyclic AMP Response Element-Binding Protein; D004745:Enkephalins; D009838:Oligodeoxyribonucleotides; D011498:Protein Precursors; D016755:Proto-Oncogene Proteins c-jun; D011994:Recombinant Proteins; D018808:Transcription Factor AP-1; D005576:Colforsin; C033660:preproenkephalin; D015500:Chloramphenicol O-Acetyltransferase