Characterization of cytokine LD78 gene promoters: Positive and negative transcriptional factors bind to a negative regulatory element common to LD78, interleukin-3, and granulocyte-macrophage colony-stimulating factor gene promoters

Hisayuki Nomiyama, Kunio Hieshima, Kaoru Hirokawa, Toshio Hattori, Kiyoshi Takatsuki, Retsu Miura

Research output: Contribution to journalArticlepeer-review

29 Citations (Scopus)

Abstract

Cytokine LD7S is a human counterpart of the mouse macrophage inflammatory protein 1α/hematopoietic stem cell inhibitor. Promoters of the LD78α and LD78β genes showed similar inducible activities in two leukemic cell lines, K562 and Jurkat, but the induction mechanisms differed between the two cell lines. Further characterization of the LD78α promoter indicated that multiple positive and negative regulatory elements are present, some of which are differentially required for induction and repression of the promoter activity in different cells. One of the negative regulatory elements, ICK-1, functioned in both cell lines in the absence and presence of stimulation and was shown to be a recognition site for positive and negative transcriptional factors. This ICK-1 element contained a direct repeat, and similar repeats were also found in the negative regulatory elements of hematopoietic growth factor interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) gene promoters. Nuclear extracts from K562 and Jurkat cells formed several protein-DNA complexes with the LD78α ICK-1 element, one of which was also observed with the IL-3 and GM-CSF ICK-1 elements. Results from in vivo and in vitro analyses suggested that the protein forming this complex functions as a negative factor. The binding affinity of this protein, ICK-1A, to the LD78α ICK-1 element was low and was significantly affected by the incubation temperature and the salt concentration in the binding buffer. ICK-1B, another protein bound specifically by the LD78α ICK-1 element, was shown to be a positive factor important for induction of the promoter. These results suggested that ICK-1A plays an important role in balanced expression of LD78, IL-3, and GM-CSF during hematopoietic cell growth and differentiation.

Original languageEnglish
Pages (from-to)2787-2801
Number of pages15
JournalMolecular and cellular biology
Volume13
Issue number5
DOIs
Publication statusPublished - 1993 May

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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