Regulation of the human long chain acyl-CoA dehydrogenase gene by nuclear hormone receptor transcription factors.
Mitochondrial fatty acid oxidation provides most of the energy required for myocardial function after birth. Long chain acyl-CoA dehydrogenase (LCAD) catalyzes the first step in the beta-oxidation spiral. Our objective was to define regulatory elements of the human LCAD gene required for high levels of expression in mature heart and to locate elements suppressing gene expression in the fetus. We characterized the human LCAD gene structure and used in vitro transfection into cardiomyocytes and hepatoma cells of LCAD genomic fragments fused to a reporter gene to examine the effects of putative regulatory elements on transcription. Binding of transcription factors to nuclear hormone receptor consensus DNA binding domains was studied by gel shift experiments. The 200 bp of the human LCAD gene immediately upstream of the transcription initiation site are sufficient to act as a minimal promoter for the gene and provide some tissue-specific positive regulatory elements. The region from -1800 bp to -250 bp contains elements which markedly suppress transcription, including nuclear hormone receptor response elements. The dominant interaction is with the repressor factor, chicken ovalbumin upstream promoter transcription factor. We conclude that the developmental and tissue-specific regulation of the human LCAD gene is mediated, in part, by these nuclear hormone receptor transcription factors.
Zhang
Z
Z
Department of Pediatrics, Washington University School of Medicine, St. Louis, MO, USA.
Zhou
Y
Y
Mendelsohn
N J
NJ
Bauer
G S
GS
Strauss
A W
AW
D016475:3T3 Cells; D044942:Acyl-CoA Dehydrogenase, Long-Chain / Q000096:biosynthesis* / Q000235:genetics*; D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D051838:COUP Transcription Factor I; D006528:Carcinoma, Hepatocellular; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D016384:Consensus Sequence; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000378:metabolism; D005091:Exons; D015971:Gene Expression Regulation, Enzymologic; D006352:Heart Ventricles; D006801:Humans; D007438:Introns; D008113:Liver Neoplasms; D051379:Mice; D009206:Myocardium / Q000166:cytology; D012326:RNA Splicing; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D014157:Transcription Factors / Q000378:metabolism*; D014162:Transfection
D051838:COUP Transcription Factor I; D017931:DNA Primers; D004268:DNA-Binding Proteins; C497589:NR2F1 protein, human; C497590:Nr2f1 protein, mouse; C497591:Nr2f1 protein, rat; D018160:Receptors, Cytoplasmic and Nuclear; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D044942:Acyl-CoA Dehydrogenase, Long-Chain; D015500:Chloramphenicol O-Acetyltransferase