The Human Medium Chain Acyl-CoA Dehydrogenase Gene Promoter Consists of a Complex Arrangement of Nuclear Receptor Response Elements and Sp1 Binding Sites (*)
- Teresa C. Leone(1),
- Sharon Cresci(1),
- M. Eric Carter(1),
- Zhifang Zhang(3),
- Deepak S. Lala(4),
- Arnold W. Strauss(3)(2) and
- Daniel P. Kelly(1)(2)(§)
- From the (1)Departments of Medicine,
- (2)Molecular Biology and Pharmacology, and
- (3)Pediatrics, Washington University School of Medicine, St. Louis, Missouri 63110 and the
- (4)Department of Medicine, Duke University Medical Center, Durham, North Carolina 27710
- § A Lucille P. Markey Scholar. To whom correspondence should be addressed: Cardiovascular Division, Box 8086, Washington University School of Medicine, 660 S. Euclid Ave., St. Louis, MO 63110. Tel.: 314-362-8919; Fax: 314-362-0186.
Abstract
Expression of the gene encoding the mitochondrial fatty acid β-oxidation enzyme, medium-chain acyl-CoA dehydrogenase (MCAD), is regulated among tissues during development and in response to alterations in substrate availability. To identify and characterize cis-acting MCAD gene promoter regulatory elements and corresponding transcription factors, DNA-protein binding studies and mammalian cell transfection analyses were performed with human MCAD gene promoter fragments. DNA:protein binding studies with nuclear protein extracts prepared from hepatoma G2 cells, 3T3 fibroblasts, or Y-1 adrenal tumor cells identified three sequences (nuclear receptor response element 1 or NRRE-1, NRRE-2, and NRRE-3) that bind orphan members of the steroid/thyroid nuclear receptor superfamily including chicken ovalbumin upstream promoter transcription factor and steroidogenic factor 1. Sp1 binding sites (A-C) were identified in close proximity to each of the NRREs. NRRE-3 conferred cell line-specific transcriptional repression by interacting with chicken ovalbumin upstream promoter transcription factor or activation via steroidogenic factor 1. In contrast, the Sp1 binding site A behaved as a transcriptional activator in all cell lines examined. We propose that multiple nuclear receptor transcription factors interact with MCAD gene promoter elements to differentially regulate transcription among a variety of cell types.
Footnotes
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↵* This work was supported by the Markey Charitable Trust and National Institutes of Health Grant DK45416-03. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵1 The abbreviations used are:
- MCAD
-
medium chain acyl-CoA dehydrogenase
- CAT
-
chloramphenicol acetyltransferase
- COUP-TF
-
chicken ovalbumin upstream promoter transcription factor
- EMSA
-
electrophoretic mobility shift assay
- FAO
-
fatty acid oxidation
- FTZ-F1
-
fushi tarazu factor 1
- HNF-4
-
hepatocyte nuclear factor 4
- NRRE
-
nuclear receptor response element
- SF-1
-
steroidogenic factor 1
- TK
-
thymidine kinase
- PCR
-
polymerase chain reaction.
