Transcriptional regulation of mouse mu opioid receptor gene in neuronal cells by Poly(ADP-ribose) polymerase-1

Hack Sun Choi, Cheol Kyu Hwang, Chun Sung Kim, Kyu Young Song, Ping Yee Law, Horace H. Loh, Li Na Wei

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

The pharmacological actions of morphine and morphine-like drugs such as heroin mediate primarily through the mu opioid receptor (MOR). It represents the target of the most valuable painkiller in contemporary medicine. Here we report that poly(ADP-ribose) polymerase 1 (PARP-1) binds to the double-stranded poly(C) element essential for the MOR promoter and represses promoter activity at the transcriptional level. We identified PARP-1 by affinity column chromatography using the double-stranded poly(C) element, followed by two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry. PARP-1 binding to the poly(C) sequence of the MOR gene was sequence-specific as confirmed by the supershift assay. In cotransfection studies, PARP-1 repressed the MOR promoter only when the poly(C) sequence was intact. When PARP-1 was disrupted in NS20Y cells using siRNA, transcription of the endogenous target MOR gene increased significantly. Chromatin immunoprecipitation assays showed specific binding of PARP-1 to the double-stranded poly(C) element essential for the MOR promoter. Inhibition of PARP-1's catalytic domain with 3-aminobenzamide increased endogenous MOR mRNA levels in cultured NS20Y cells, suggesting that automodification of PARP-1 regulates MOR transcription. Our data suggest that PARP-1 can function as a repressor of MOR transcription dependent on the MOR poly(C) sequence. We demonstrate for the first time a role of PARP-1 as a transcriptional repressor in MOR gene regulation.

Original languageEnglish (US)
Pages (from-to)2319-2333
Number of pages15
JournalJournal of Cellular and Molecular Medicine
Volume12
Issue number6A
DOIs
StatePublished - Dec 2008

Keywords

  • 3-aminobenzamide
  • Mu opioid receptor
  • Poly(ADP-ribose) polymerase
  • Poly(C) element
  • Transcriptional repressor

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