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

Tetsuro Orita, Koji Shimozaki, Hiroshi Murakami, Shigekazu Nagata

Research output: Contribution to journalArticle

25 Citations (Scopus)

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 acetyl-transferase 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.

Original languageEnglish
Pages (from-to)23216-23223
Number of pages8
JournalJournal of Biological Chemistry
Volume272
Issue number37
DOIs
Publication statusPublished - 1997
Externally publishedYes

Fingerprint

Granulocyte Colony-Stimulating Factor
Peroxidase
Transcription Factors
Genes
Cultured Cells
Granulocyte Colony-Stimulating Factor Receptors
Affinity chromatography
Sequence Deletion
Interleukin-3
5' Flanking Region
Transcription Initiation Site
Protein Sequence Analysis
Cell Lineage
Myeloid Cells
Chloramphenicol
Transferases
Affinity Chromatography
Oligonucleotides
Base Pairing
Assays

ASJC Scopus subject areas

  • Biochemistry

Cite this

Binding of NF-Y transcription factor to one of the Cis-elements in the myeloperoxidase gene promoter that responds to granulocyte colony-stimulating factor. / Orita, Tetsuro; Shimozaki, Koji; Murakami, Hiroshi; Nagata, Shigekazu.

In: Journal of Biological Chemistry, Vol. 272, No. 37, 1997, p. 23216-23223.

Research output: Contribution to journalArticle

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