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.

Spyrou, GiannisKarolinska Institutet, Stockholm, Sweden(Swepub:liu)ioasp42 (författare)
Rökaeus, ÅkeKarolinska Institutet (författare)
Karolinska InstitutetKarolinska Institutet, Stockholm, Sweden (creator_code:org_t)

Ingår i:Biochemical and Biophysical Research Communications - BBRC: Elsevier246:1, s. 192-1980006-291X1090-2104

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