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NF-kappaB binds to a polymorphic repressor element in the MMP-3 promoter

Ruth C Borghaei1, P Lyle Rawlings, Masoud Javadi

  • 1Department of Biochemistry and Molecular Biology, Philadelphia College of Osteopathic Medicine, 4170 City Avenue, Philadelphia, PA 19131, USA. ruthb@pcom.edu

Biochemical and Biophysical Research Communications
|March 9, 2004
PubMed

Insights

The 6T allele of the matrix metalloproteinase-3 (MMP-3) promoter may limit its expression. Nuclear factor-kappa B (NF-kappaB) binding to this site, along with ZBP-89, suggests a role in regulating MMP-3 transcription.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Signaling

Background:

  • A polymorphic site (5T/6T) in the matrix metalloproteinase-3 (MMP-3) promoter acts as a repressor element.
  • The 6T allele is linked to reduced MMP-3 expression compared to the 5T allele.
  • Zinc-binding protein-89 (ZBP-89) interacts with this site but activates the promoter upon overexpression.

Purpose of the Study:

  • To investigate the molecular mechanisms underlying MMP-3 promoter regulation by the 5T/6T polymorphism.
  • To identify proteins binding to the polymorphic site in response to interleukin-1 (IL-1) stimulation.
  • To elucidate the role of NF-kappaB and ZBP-89 in MMP-3 gene expression.

Main Methods:

  • Yeast one-hybrid assay for ZBP-89 cloning.
  • Nuclear extract analysis from IL-1-stimulated human gingival fibroblasts.
  • Electrophoretic mobility shift assays (EMSAs) to study protein-DNA interactions.
  • Recombinant protein binding studies.

Main Results:

  • The 5T/6T polymorphic site in the MMP-3 promoter is bound by p50 and p65 subunits of NF-kappaB in IL-1-stimulated cells.
  • ZBP-89 also binds to this site.
  • Recombinant p50 protein demonstrates preferential binding to the 6T allele.
  • NF-kappaB binding is consistent with a role in limiting cytokine-induced MMP-3 expression.

Conclusions:

  • NF-kappaB, in conjunction with ZBP-89, binds to the MMP-3 promoter's polymorphic site.
  • The preferential binding of p50 to the 6T allele suggests a mechanism for allele-specific regulation of MMP-3 expression.
  • These findings support a role for NF-kappaB in modulating MMP-3 transcription in response to inflammatory signals.

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