Nucleic Acids Research, 1994, Vol. 22, No. 11 2126-2133
© 1994
MOLECULAR BIOLOGY |
Retinoic acid-mediated activation of HNF-3
during EC stem cell differentiation
Department of Pharmacology and Molecular Biology, The Chicago Medical School North Chicago, IL 60064 1Department of Biochemistry, University of Illinois, College of Medicine Chicago, IL 60612, USA
*To whom correspondence should be addressed
Received January 19, 1994. Revised May 5, 1994. Accepted May 5, 1994.
We present evidence demonstrating that the liver-enriched transcription factor HNF-3
is activated upon retinoic acid-induced differentiation of mouse F9 embryonal carcinoma cells. We have detected increases in the DNA binding activity and mRNA level of HNF-3. Both are reflections of the actual activation mechanism at the level of transcriptional initiation, which we showed with the help of HNF-3 promoter constructs. Time course studies clearly show that HNF-3 activation is a transient event. Employing Northern blots, HNF-3 mRNA can be detected between 16 and 24 hours post-differentiation, reaches its zenith at approximately 1 day, and then declines to virtually undetectable levels. F9 cells can give rise to three distinct differentiated cell types; visceral endoderm, parietal endoderm, and primitive endoderm. We have clearly shown that HNF-3 stimulation occurs upon primitive endoderm formation. In addition, the transcription factor is also activated during the induction of cell lineages that give rise to parietal and visceral endoderm. HNF-3 stimulation upon visceral endoderm differentiation is accompanied by the activation of HNF-3 target genes such as transthyretin, suggesting that HNF-3 is involved in the developmental activation of this gene. In contrast, HNF-3 target genes in parietal and primitive endoderm have yet to be identified. However, the stimulation of HNF-3 during primitive endoderm formation, which is an extremely early event during murine embryogenesis, points towards a role for the factor in crucial determination processes that occur early during development.
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