Binding of NF-Y transcription factor to one of the cis-elements in the myeloperoxidase gene promoter that responds to granulocyte colony-stimulating factor

J Biol Chem. 1997 Sep 12;272(37):23216-23. doi: 10.1074/jbc.272.37.23216.

Abstract

The expression of the myeloperoxidase (MPO) gene is restricted to cells of the myeloid cell lineage and is induced by granulocyte colony-stimulating factor (G-CSF). In this study, a series of deletion mutations was introduced in the promoter of the human MPO gene, which was then fused to the chloramphenicol acetyltransferase gene. The G-CSF-induced promoter activity was examined in mouse myeloid precursor FDC-P1 transformants that constitutively express the G-CSF receptor. A G-CSF-responsive element (GRE) in the MPO gene was found approximately 800 base pairs upstream from the transcription initiation site. When the 5'-flanking region of the human MPO gene contained this element, it yielded promoter activity in cells cultured with G-CSF but not in cells cultured with interleukin 3. Gel shift assays with the element showed that a specific nuclear factor(s) (NF/G-CSF) binds to the element. The NF/G-CSF was purified by affinity chromatography using an oligonucleotide of GRE. Protein sequence analysis of the purified NF/G-CSF indicated that NF/G-CSF is a ubiquitous transcription factor, NF-Y, which is composed of three subunits. The recombinant NF-Y was then shown to bind to GRE in a combination of the three subunits.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites
  • CCAAT-Enhancer-Binding Proteins
  • DNA-Binding Proteins / metabolism*
  • Gene Expression Regulation, Enzymologic*
  • Granulocyte Colony-Stimulating Factor / metabolism*
  • Humans
  • Interleukin-3 / metabolism
  • Mice
  • Molecular Sequence Data
  • Peroxidase / genetics*
  • Promoter Regions, Genetic*
  • Protein Binding
  • Recombinant Proteins
  • Sequence Analysis
  • Sequence Deletion
  • Transcription Factors / metabolism*
  • Transformation, Genetic

Substances

  • CCAAT-Enhancer-Binding Proteins
  • DNA-Binding Proteins
  • Interleukin-3
  • Recombinant Proteins
  • Transcription Factors
  • Granulocyte Colony-Stimulating Factor
  • Peroxidase

Associated data

  • GENBANK/AB006181