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Vol. 61, Issue 4, 800-805, April 2002

Involvement of Activator Protein-1 in Transcriptional Regulation of the Human µ-Opioid Receptor Gene

Christine Börner, Volker Höllt, and Jürgen Kraus

Department of Pharmacology, University of Magdeburg, Magdeburg, Germany

µ-Opioid receptors mediate such opioid effects as analgesia, euphoria, and immunomodulation. Gene expression of µ-opioid receptors can be modulated by various substances, including cytokines, hormones, and drugs. Some of these stimuli (e.g., IL-1beta and cocaine) have been shown to activate members of the AP-1 transcription factor family. In addition, transcription of the µ-opioid receptor gene is induced by the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), an activator of protein kinase C, which in turn is an activator of AP-1 transcription factors. This indicates that signaling pathways involving protein kinase C and activator protein 1 (AP-1) transcription factors are important for the specific expression pattern of the µ-opioid receptor gene. In this report, we show that TPA activates AP-1 as well as the transcription factor nuclear factor kappa B (NFkappa B) in the µ-opioid receptor expressing neuroblastoma cell line SH SY5Y. In transfection experiments performed in these cells, both factors trans-activate expression of reporter gene constructs containing the human µ-opioid receptor gene promoter. By excluding the effects of TPA on NFkappa B with the specific NFkappa B inhibitor sulfasalazine, AP-1 regulatory elements were localized. Two AP-1 elements, which differ in one nucleotide each from the classic AP-1 binding site, were delineated to positions -2388 and -1434 of the promoter. Independent of their orientation, these elements conferred TPA responsiveness on the heterologous thymidine kinase promoter. AP-1 binding to these elements was confirmed using electrophoretic mobility shift and immunoshift assays.

Copyright © 2002 by The American Society for Pharmacology and Experimental Therapeutics


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Copyright © 2002 by the American Society for Pharmacology and Experimental Therapeutics