GATA suppresses erythropoietin gene expression through GATA site in mouse erythropoietin gene promoter.
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 with luciferase. 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), promoterluciferase construct but not the hypoxic induction of the mutant (m4, 5) promoter-luciferase constructs. N(G)-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.
Imagawa
Shigehiko
S
Division of Hematology, Institute of Clinical Medicine, University of Tsukuba, Ibaraki, Japan. simagawa@md.tsukuba.ac.jp
Suzuki
Norio
N
Ohmine
Ken
K
Obara
Naoshi
N
Mukai
Harumi Y
HY
Ozawa
Keiya
K
Yamamoto
Masayuki
M
Nagasawa
Toshiro
T
D000818:Animals; D001120:Arginine / Q000494:pharmacology; D001665:Binding Sites; D002374:Catalase / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000235:genetics / Q000494:pharmacology* / Q000502:physiology; D015536:Down-Regulation / Q000187:drug effects; D050981:Erythroid-Specific DNA-Binding Factors; D004921:Erythropoietin / Q000235:genetics* / Q000378:metabolism; D050989:GATA2 Transcription Factor; D050990:GATA3 Transcription Factor; D006861:Hydrogen Peroxide / Q000494:pharmacology; D051379:Mice; D011401:Promoter Regions, Genetic / Q000187:drug effects* / Q000235:genetics; D015534:Trans-Activators / Q000235:genetics / Q000494:pharmacology / Q000502:physiology; D014157:Transcription Factors / Q000235:genetics / Q000494:pharmacology* / Q000502:physiology; D014162:Transfection; D014407:Tumor Cells, Cultured; D019323:omega-N-Methylarginine / Q000494:pharmacology
D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D050989:GATA2 Transcription Factor; D050990:GATA3 Transcription Factor; C494712:Gata2 protein, mouse; C494748:Gata3 protein, mouse; D015534:Trans-Activators; D014157:Transcription Factors; D004921:Erythropoietin; D019323:omega-N-Methylarginine; D001120:Arginine; D006861:Hydrogen Peroxide; D002374:Catalase