Abstract
The ability of peroxisome proliferator-activated receptors (PPARs) to induce expression of a reporter gene linked to a peroxisome proliferator-responsive element (PPRE) from either the rat enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase gene or acyl-CoA oxidase [acyl-CoA:oxygen 2-oxidoreductase, EC 1.3.3.6] gene was examined by transient transfection assays in COS cells. Mouse and rat PPARs, as well as Xenopus PPAR alpha (xPPAR alpha) could induce expression of a reporter gene linked to the hydratase/dehydrogenase PPRE in the presence of the peroxisome proliferators ciprofibrate or Wy-14,643, whereas xPPAR beta and xPPAR gamma were ineffective. A similar induction of expression of a reporter gene linked to the acyl-CoA oxidase PPRE was observed with all PPARs except xPPAR beta. Extracts from cells transfected with PPAR-encoding genes contained factors that bound to both PPREs. In vitro synthesized PPARs could interact weakly with both PPREs; however, binding of each PPAR to both PPREs was significantly increased by the addition of COS cell nuclear extracts, demonstrating that efficient PPAR/DNA binding requires auxiliary cofactors. One cofactor was identified as the 9-cis-retinoic acid receptor, RXR alpha (retinoid X receptor alpha). Cooperative DNA binding and heteromerization between RXR alpha and each of the PPARs could be seen with both PPREs. Our results demonstrate that PPAR/PPRE binding and cooperativity with RXR alpha (and other cofactors) are obligatory but not necessarily sufficient for peroxisome proliferator-dependent transcription induction and that distinct PPREs can selectively mediate induction by particular PPARs.
MeSH Terms
3-Hydroxyacyl CoA Dehydrogenases/genetics
Acyl-CoA Oxidase
Animals
Base Sequence
Binding Sites
DNA/metabolism
Enoyl-CoA Hydratase/genetics
Gene Expression Regulation, Enzymologic
Isomerases/genetics
Kinetics
Liver/enzymology
Liver Neoplasms, Experimental
Mice
Molecular Sequence Data
Multienzyme Complexes/genetics
Oligodeoxyribonucleotides
Oxidoreductases/genetics
Peroxisomal Bifunctional Enzyme
Rats
Receptors, Cell Surface/metabolism
Receptors, Cytoplasmic and Nuclear
Restriction Mapping
Transcription Factors/metabolism
Transfection
Tumor Cells, Cultured
Xenopus
Chemicals
Multienzyme Complexes
Oligodeoxyribonucleotides
Receptors, Cell Surface
Receptors, Cytoplasmic and Nuclear
Transcription Factors
DNA
Oxidoreductases
3-Hydroxyacyl CoA Dehydrogenases
Acyl-CoA Oxidase
Ehhadh protein, mouse
Enoyl-CoA Hydratase
Peroxisomal Bifunctional Enzyme
Isomerases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Marcus S L
Department of Biochemistry, McMaster University, Hamilton, ON, Canada.
Miyata K S
Zhang B
Subramani S
Rachubinski R A
Capone J P
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