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Volume 272, Number 37, Issue of September 12, 1997 pp. 23216-23223
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Binding of NF-Y Transcription Factor to One of the Cis-elements in the Myeloperoxidase Gene Promoter That Responds to Granulocyte Colony-stimulating Factor

(Received for publication, August 16, 1996, and in revised form, June 23, 1997)

Tetsuro Orita , Koji Shimozaki ^¶ , Hiroshi Murakami and Shigekazu Nagata ^¶

From the  Osaka Bioscience Institute, 6-2-4 Furuedai, Suita, Osaka 565, Japan and the ^¶ Department of Genetics, Osaka University Medical School, 2-2 Yamadaoka, Suita, Osaka 565, Japan

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.

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