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Identification of an NF-κB p50/p65-responsive site in the human MIR155HG promoter

Ryan C Thompson1, Iosif Vardinogiannis, Thomas D Gilmore

  • 1Department of Biology, Boston University, Boston, MA 02215, USA. gilmore@bu.edu.

BMC Molecular Biology
|September 25, 2013
PubMed
Abstract

Insights

MicroRNA-155 (miR-155) is directly regulated by the NF-κB pathway, as demonstrated by identifying a specific NF-κB-responsive element in the MIR155HG promoter. This finding advances understanding of immune cell regulation and potential cancer therapies.

Area of Science:

  • Molecular Biology
  • Immunology
  • Genetics

Background:

  • MicroRNA-155 (miR-155), derived from the MIR155HG gene, plays a crucial role in immune cell regulation.
  • Aberrant miR-155 expression is linked to human lymphomas and oncogenic activity.
  • Previous studies suggested a correlation between NF-κB activity and MIR155HG expression, but direct promoter regulation was unproven.

Purpose of the Study:

  • To investigate the direct regulatory role of the transcription factor NF-κB on the human MIR155HG gene promoter.
  • To identify and characterize the specific DNA elements involved in NF-κB-mediated regulation of MIR155HG.

Main Methods:

  • Induction of NF-κB activity in cells.
  • Quantification of primary MIR155HG mRNA and mature miR-155 transcripts.
  • Mapping of NF-κB-responsive elements using promoter analysis.
  • Electrophoretic mobility shift assays (EMSA) to confirm protein binding.
  • Reporter gene assays to assess promoter activity.
  • Correlation analysis of miR-155 levels and nuclear NF-κB proteins in lymphoma cell lines.

Main Results:

  • NF-κB activation rapidly increased both MIR155HG mRNA and miR-155 levels.
  • An NF-κB-responsive element was identified 178 nucleotides upstream of the MIR155HG transcription start site (-178 site).
  • The -178 site specifically binds the NF-κB p50/p65 heterodimer and is essential for p65-induced reporter gene activation.
  • miR-155 levels correlated with nuclear NF-κB protein levels in human B-lymphoma cell lines.

Conclusions:

  • The MIR155HG gene is a direct target of NF-κB, confirmed by the identification of a functional NF-κB-responsive site in its promoter.
  • This establishes NF-κB-mediated transcriptional control over miR-155 production.
  • Understanding this regulatory pathway may facilitate the development of novel diagnostic tools and targeted therapies for immune-related diseases and lymphomas.

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