Cooperative Interaction between Hepatocyte Nuclear Factor 4α and GATA Transcription Factors Regulates ATP-Binding Cassette Sterol Transporters ABCG5 and ABCG8

Abstract

Cholesterol homeostasis is maintained by coordinate regulation of cholesterol synthesis and its conversion to bile acids in the liver. The excretion of cholesterol from liver and intestine is regulated by ATP-binding cassette half-transporters ABCG5 and ABCG8. The genes for these two proteins are closely linked and divergently transcribed from a common intergenic promoter region. Here, we identified a binding site for hepatocyte nuclear factor 4α (HNF4α) in the ABCG5/ABCG8 intergenic promoter, through which HNF4α strongly activated the expression of a reporter gene in both directions. The HNF4α-responsive element is flanked by two conserved GATA boxes that were also required for stimulation by HNF4α. GATA4 and GATA6 bind to the GATA boxes, coexpression of GATA4 and HNF4α leads to a striking synergistic activation of both the ABCG5 and the ABCG8 promoters, and binding sites for HNF4α and GATA were essential for maximal synergism. We also show that HNF4α, GATA4, and GATA6 colocalize in the nuclei of HepG2 cells and that a physical interaction between HNF4α and GATA4 is critical for the synergistic response. This is the first demonstration that HNF4α acts synergistically with GATA factors to activate gene expression in a bidirectional fashion.

We thank Johan Auwerx, Takashi Minami, and Makoto Makishima for helpful discussions.

This study was supported in part by grants from the Program of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation, by the NFAT project of New Energy and Industrial Technology Development Organization, and by the Special Coordination Fund for Science and Technology from the Ministry of Education, Culture, Sports, Science, and Technology, the Uehara Memorial foundation, and the Ono Medical foundation. J.S. is a recipient of funds from the Special Coordination Fund for Science and Technology from the Ministry of Education, Culture, Sports, Science and Technology.