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J Biol Chem, Vol. 274, Issue 12, 8231-8239, March 19, 1999
From the Department of Cardiovascular Medicine, University of Tokyo
Graduate School of Medicine, 7-3-1 Hongo, Bunkyo-ku,
Tokyo 113-8655, Japan
Although the cardiac homeobox gene
Csx/Nkx-2.5 is essential for normal heart development,
little is known about its regulatory mechanisms. In a search for the
downstream target genes of Csx/Nkx-2.5, we found that the atrial
natriuretic peptide (ANP) gene promoter was strongly transactivated by
Csx/Nkx-2.5. Deletion and mutational analyses of the ANP promoter
revealed that the Csx/Nkx-2.5-binding element (NKE2) located at 
240
was required for high level transactivation by Csx/Nkx-2.5. We also
found that Csx/Nkx-2.5 and GATA-4 displayed synergistic transcriptional
activation of the ANP promoter, and in contrast to previous reports
(Durocher, D., Charron, F., Warren, R., Schwartz, R. J., and
Nemer, M. (1997) EMBO J. 16, 5687-5696; Lee, Y., Shioi,
T., Kasahara, H., Jobe, S. M., Wiese, R. J., Markham, B., and
Izumo, S (1998) Mol. Cell. Biol. 18, 3120-3129), this synergism was dependent on binding of Csx/Nkx-2.5 to NKE2, but not on
GATA-4-DNA interactions. Although GATA-4 also potentiated the
Csx/Nkx-2.5-induced transactivation of the artificial promoter that
contains multimerized Csx/Nkx-2.5-binding sites, Csx/Nkx-2.5 reduced
the GATA-4-induced transactivation of the GATA-4-dependent promoters. These findings indicate that the cooperative transcriptional regulation mediated by Csx/Nkx-2.5 and GATA-4 is promoter
context-dependent and suggest that the complex
cis-trans interactions may fine-tune gene
expression in cardiac myocytes.
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