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Poly(C) binding protein family is a transcription factor in mu-opioid receptor gene expression

Sung-Su Kim1, Krishan K Pandey, Hack Sun Choi

  • 1Department of Pharmacology, University of Minnesota Medical School, 6-120 Jackson Hall, 321 Church St. SE, Minneapolis, MN 55455, USA. kimxx558@umn.edu

Molecular Pharmacology
|June 4, 2005
PubMed

Insights

Poly(C) binding proteins (PCBPs) interact with the mouse mu-opioid receptor (MOR) gene's promoter, activating MOR transcription. These transcription factors enhance MOR gene expression in NMB cells.

Area of Science:

  • Molecular Biology
  • Neuroscience
  • Gene Regulation

Background:

  • The mouse mu-opioid receptor (MOR) gene possesses two promoters: distal and proximal.
  • A previously identified 26-base pair (bp) cis-acting element in the mouse MOR gene activates MOR gene expression.

Purpose of the Study:

  • To identify proteins interacting with the 26-bp polypyrimidine stretch in the MOR proximal promoter.
  • To elucidate the role of these interacting proteins in regulating MOR gene transcription.

Main Methods:

  • Cloning of four poly(C) binding protein (PCBP) family members.
  • Nuclear run-off assays and semiquantitative RT-PCR to assess transcription rates.
  • Chromatin immunoprecipitation assays to confirm in vivo interactions.

Main Results:

  • Four PCBP family members were cloned and shown to interact with the 26-bp polypyrimidine stretch in the MOR proximal promoter.
  • PCBPs bind to both single- and double-stranded DNA, enhancing MOR gene transcription rates.
  • Refined mapping identified a core region (-317/-304) crucial for PCBP-induced MOR promoter activity, confirmed by decoy oligonucleotides.

Conclusions:

  • Poly(C) binding proteins (PCBPs) function as transcription factors that positively regulate mouse mu-opioid receptor (MOR) gene expression.
  • The interaction between PCBPs and the MOR promoter is essential for MOR gene transcription in NMB cells.

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