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Blood, 1 November 2000, Vol. 96, No. 9, pp. 3265-3271
RED CELLS
Retinoic acid stimulates erythropoietin gene transcription in
embryonal carcinoma cells through the direct repeat of a
steroid/thyroid hormone receptor response element half-site in the
hypoxia-response enhancer
Taiho Kambe,
Junko Tada-Kambe,
Yoshihiro Kuge,
Yuko Yamaguchi-Iwai,
Masaya Nagao, and
Ryuzo Sasaki
From the Division of Integrated Life Science, Graduate
School of Biostudies, Kyoto University, Kyoto, Japan.
We have previously reported that expression of the
erythropoietin (Epo) gene in mouse embryonal cells was not induced by
hypoxia, although hypoxia induced other hypoxia-inducible genes. This
study identifies retinoic acid (RA) as an inducer for Epo production in
the embryonal carcinoma cell lines P19 and F9. RA induced Epo production through the transcriptional activation of the Epo gene in an
oxygen-independent manner. With the use of reporter assays in P19
cells, it is shown that a direct repeat of the nuclear hormone
receptor-binding motif separated by a 2-bp spacer (DR-2) in the
hypoxia-response enhancer was responsible for the transcriptional activation by RA. Electrophoretic mobility shift assays show that nuclear extracts from P19 cells contained RA receptor complexes that
bound to DR-2. In human hepatoma Hep3B cells, an orphan receptor, hepatocyte nuclear factor-4, strongly augmented hypoxic induction of
the Epo gene in cooperation with hypoxia-inducible factor-1 (HIF-1) by
binding to DR-2, whereas in P19 cells, the interaction of RA receptors
with DR-2 was sufficient for RA-induced transcriptional activation of
the Epo gene without the requirement of the HIF-1 site. These results
suggest that DR-2 regulates expression of the Epo gene by acting as the
binding site for different transcription factors in different types of cells.
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