International Journal of Hematology

, Volume 75, Issue 4, pp 376–381 | Cite as

GATA Suppresses Erythropoietin Gene Expression through GATA Site in Mouse Erythropoietin Gene Promoter

  • Shigehiko Imagawa
  • Norio Suzuki
  • Ken Ohmine
  • Naoshi Obara
  • Harumi Y. Mukai
  • Keiya Ozawa
  • Masayuki Yamamoto
  • Toshiro Nagasawa
Review Article

Abstract

The promoter and enhancer elements of the mouse erythropoietin (mEpo) gene, which have high homology with those of the human erythropoietin (hEpo) gene, were fused withluciferase. The construct was transfected into erythropoietin-producing hepatoma cell line (Hep3B) cells by lipofectin with lacZ as an internal standard. The wild type (TGATA) showed a 39.5-fold increase in induction by hypoxia. Mouse GATA-2 inhibited the hypoxic induction of the wild-type (m3), promoter-luciferase construct but not the hypoxic induction of the mutant (m4, 5) promoter-luciferase constructs. NG-monomethyl L-arginine (L-NMMA) inhibited the hypoxic induction of the m3 promoter-luciferase construct, but this inhibition was recovered by L-arginine. H2O2 also inhibited the hypoxic induction of the m3 promoter-luciferase construct, but this inhibition was recovered by catalase. Gel shift assays performed on nuclear extracts of 293 cells overexpressing mGATA-1, -2, and -3 revealed that mGATA-1, -2, and -3 bind to the TGATA element of the mEpo promoter. These results indicate that mGATA binds to the TGATA site of the mEpo promoter and negatively regulates mEpo gene expression. Negative regulation of mEpo gene by GATA transcriptional factors is discussed.

Key words

Erythropoietin GATA Transcriptional regulation L-NMMA H2O2 

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Copyright information

© The Japanese Society of Hematology 2002

Authors and Affiliations

  • Shigehiko Imagawa
    • 1
  • Norio Suzuki
    • 2
  • Ken Ohmine
    • 3
  • Naoshi Obara
    • 1
  • Harumi Y. Mukai
    • 1
  • Keiya Ozawa
    • 3
  • Masayuki Yamamoto
    • 2
  • Toshiro Nagasawa
    • 1
  1. 1.Division of Hematology, Institute of Clinical MedicineUniversity of TsukubaIbarakiJapan
  2. 2.Center for Tsukuba Advanced Research Alliance and Institute of Basic Medical SciencesUniversity of TsukubaIbaraki
  3. 3.Department of HematologyJichi Medical SchoolTochigiJapan

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