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Negative regulation of the erythropoietin gene expression by the GATA
transcription factors
S Imagawa, M Yamamoto and Y Miura
Department of Medicine, Jichi Medical School, Tochigi-ken, Japan.
We examined regulation of the human erythropoietin (Epo) gene through the
GATA sequence in the Epo promoter and showed that Hep3B and HepG2 cells
express human GATA-2 (hGATA-2) mRNA and protein. Nuclear extracts of QT6
cells transfected with hGATA-1, 2, or 3 transcription factors showed
specific binding to the GATA element in the human Epo gene promoter by gel
mobility shift assay. Transient transfection of Hep3B cells with hGATA-1,
2, or 3 showed that each of these transcription factors significantly
decreased the level of expression of Epo mRNA as assessed by a competitive
polymerase chain reaction. Transient transfection of Hep3B cells with
hGATA-1, 2, and 3 and an Epo-reporter gene (growth hormone [GH]) construct
showed significant inhibition of the Epo promoter. Antisense
oligonucleotide for hGATA-2 transcription factor significantly increased
the Epo protein in Hep3B cells under 1% O2 for 24 hours incubation.
Furthermore, transient transfection of Hep3B cells with hGATA-1, 2, and 3
and an Epo-reporter gene (luciferase) construct also showed significant
inhibition of the Epo promoter. However, transfection of the mutated GATA
sequence of the Epo- luciferase gene with hGATA-1, 2, and 3 interfere with
the inhibition of the Epo promoter. We conclude that the hGATA-1, 2, and 3
transcription factors specifically bind to the GATA element in the human
Epo gene promoter and negatively regulate Epo gene expression.
Volume 89,
Issue 4,
pp. 1430-1439,
02/15/1997
Copyright © 1997 by The American Society of Hematology
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