Characterization of phorbolester-inducible human neuronal factors involved in trans-activation of the galanin gene.
The expression of the neuropeptide galanin (GAL) is elevated in vivo upon nerve stimulation, injury, and in vitro by phorbol 12-myristate-13-acetate (PMA), suggesting that a signal pathway involving protein kinase C activation may be involved in GAL-gene activation. When plasmids containing a different length of the bovine GAL-promoter fused to luciferase were transfected into the human neuroblastoma cell line (SK-N-SH subclone SH-SY5Y), a PMA-responsive element was identified in the promoter-region -68 to -46 base pairs (bp). Co-transfection experiments with plasmids expressing cJun and cFos revealed that they could act alone, as well as synergistically with PMA to induce luciferase activity. Electrical mobility shift assays revealed that a cAMP response element (CRE)-like sequence (TGACGCGG; -59 to -52 bp) bound PMA-inducible nuclear proteins present in SH-SY5Y cells. These proteins appear to bind mainly as CRE-binding protein/activating-transcription-factor (CREB/ATF) and Jun/ATF heterodimers. In addition, an apparent PMA-inducible protein(s) not recognized by CREB/ATF and Jun antibodies bound to the CRE-like containing probe.
Jiang
K
K
Department of Medical Biochemistry and Biophysics (MBB), Karolinska Institutet, Stockholm, Sweden.
Spyrou
G
G
Rökaeus
A
A
D051698:Activating Transcription Factor 2; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002999:Clone Cells; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D019004:Galanin / Q000235:genetics*; D016762:Genes, fos; D016758:Genes, jun; D006801:Humans; D008156:Luciferases / Q000235:genetics; D009479:Neuropeptides / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D011401:Promoter Regions, Genetic; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects*; D014162:Transfection
D051698:Activating Transcription Factor 2; D017362:Cyclic AMP Response Element-Binding Protein; D009479:Neuropeptides; D009838:Oligodeoxyribonucleotides; D018808:Transcription Factor AP-1; D014157:Transcription Factors; D019004:Galanin; D008156:Luciferases; D013755:Tetradecanoylphorbol Acetate