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Journal of Lipid Research, Vol. 41, 1-11, January 2000
Copyright © 2000 by Lipid Research, Inc.

Original Article

HNF4 and COUP-TFII interact to modulate transcription of the cholesterol 7{alpha}-hydroxylase gene (CYP7A1)

Diane Stroupa and John Y. L. Chianga
a Department of Biochemistry and Molecular Pathology, Northeastern Ohio Universities College of Medicine, P. O. Box 95, Rootstown, OH 44272-0095

Correspondence to: John Y. L. Chiang

The gene for cholesterol 7{alpha}-hydroxylase (CYP7A1) contains a sequence at nt -149 to -118 that was found to play a large role in determining the overall transcriptional activity and regulation of the promoter. Hepatocyte nuclear factor 4 (HNF4) and chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) synergistically activate transcription of the CYP7A1 promoter. Transactivation of CYP7A1 by HNF4 in the human hepatoma cell line, HepG2, was enhanced by cotransfection with COUP-TFII or the basal transcription element binding protein (BTEB). HNF4 prepared from rat liver nuclear extracts bound to oligomers homologous to the nt -146 to -134 sequences in electrophoretic mobility shift assays (EMSA), which corresponded to a conserved region containing a direct repeat of hormone response elements spaced by one nucleotide (DR1). The sequences surrounding this DR1 were found to be essential for the HNF4 transactivation. In vitro-translated COUP-TFII was found to bind the adjacent sequences from nt -139 to -128 (DR0), but COUP-TFII interacted with this region at a much lower affinity than to the COUP-TFII-site at nt -72 to -57 (DR4). Mutations at nt -139 to -128 or nt -72 to -57 reduced the COUP-TFII and HNF4 synergy; however, these COUP-TFII-binding sequences were not absolutely required for the cooperative effect of HNF4 and COUP-TFII on transactivation.

These results indicated that the observed transactivation was the result of protein/protein interactions facilitated by the juxtaposition of the binding elements.—Stroup, D., and J. Y. L. Chiang. HNF4 and COUP-TFII interact to modulate transcription of the cholesterol 7{alpha}-hydroxylase gene (CYP7A1). J. Lipid Res. 2000. 41: 1;–11.

Supplementary key words: bile acid synthesis, hepatocyte nuclear factor 4, chicken ovalbumin upstream promoter transcription factor II, orphan receptors, transcriptional regulation, cholesterol metabolism


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