Induction of human matrix metalloproteinase-12 gene transcriptional activity by GM-CSF requires the AP-1 binding site in human U937 monocytic cells

Biochem Biophys Res Commun. 2001 Jul 13;285(2):300-7. doi: 10.1006/bbrc.2001.5161.

Abstract

Matrix metalloproteinase-12 (MMP-12) is critical for the migration of monocytes/macrophages into inflammatory sites through the basement membranes. We previously reported that MMP-12 expression was initially induced by granulocyte macrophage colony-stimulating factor (GM-CSF) in human peripheral blood monocytes and U937 monocytic cells. To further elucidate the molecular mechanism for the regulation of MMP-12 expression by GM-CSF in monocytes, we determined the sequence requirements for the MMP-12 gene transcriptional response of U937 monocytic cells to GM-CSF by using luciferase reporter and electrophoretic mobility shift assays. A series of 5'-deletion and site-directed mutation of the human MMP-12 promoter demonstrated that an AP-1 site spanning the -81 to -75-bp region is critical for the induction of MMP-12 promoter activity by GM-CSF. The electrophoretic mobility shift assay revealed that AP-1 binding activity was increased by GM-CSF treatment and that the AP-1 complex induced by GM-CSF consisted of multiple Jun and Fos isoforms. These results indicate that MMP-12 expression in U937 monocytes was initially induced by GM-CSF through the AP-1 binding activity.

MeSH terms

  • 5' Untranslated Regions / genetics
  • Binding Sites
  • Gene Expression Regulation, Enzymologic / drug effects
  • Gene Expression Regulation, Enzymologic / physiology*
  • Genes, Reporter
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology*
  • Humans
  • Luciferases / genetics
  • Matrix Metalloproteinase 12
  • Metalloendopeptidases / genetics*
  • Monocytes / enzymology*
  • Polymerase Chain Reaction
  • Promoter Regions, Genetic*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Sequence Deletion
  • TATA Box
  • Transcription Factor AP-1 / chemistry
  • Transcription Factor AP-1 / metabolism*
  • Transcription, Genetic / drug effects
  • Transcription, Genetic / physiology*
  • Transfection
  • U937 Cells

Substances

  • 5' Untranslated Regions
  • Recombinant Proteins
  • Transcription Factor AP-1
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Luciferases
  • Metalloendopeptidases
  • MMP12 protein, human
  • Matrix Metalloproteinase 12