@article{18a3c900ba9611dc9626000ea68e967b,
title = "Acyl-CoA esters antagonize the effects of ligands on peroxisome proliferator-activated receptor alpha conformation, DNA binding, and interaction with Co-factors.",
abstract = "The peroxisome proliferator-activated receptor alpha (PPARalpha) is a ligand-activated transcription factor and a key regulator of lipid homeostasis. Numerous fatty acids and eicosanoids serve as ligands and activators for PPARalpha. Here we demonstrate that S-hexadecyl-CoA, a nonhydrolyzable palmitoyl-CoA analog, antagonizes the effects of agonists on PPARalpha conformation and function in vitro. In electrophoretic mobility shift assays, S-hexadecyl-CoA prevented agonist-induced binding of the PPARalpha-retinoid X receptor alpha heterodimer to the acyl-CoA oxidase peroxisome proliferator response element. PPARalpha bound specifically to immobilized palmitoyl-CoA and Wy14643, but not BRL49653, abolished binding. S-Hexadecyl-CoA increased in a dose-dependent and reversible manner the sensitivity of PPARalpha to chymotrypsin digestion, and the S-hexadecyl-CoA-induced sensitivity required a functional PPARalpha ligand-binding pocket. S-Hexadecyl-CoA prevented ligand-induced interaction between the co-activator SRC-1 and PPARalpha but increased recruitment of the nuclear receptor co-repressor NCoR. In cells, the concentration of free acyl-CoA esters is kept in the low nanomolar range due to the buffering effect of high affinity acyl-CoA-binding proteins, especially the acyl-CoA-binding protein. By using PPARalpha expressed in Sf21 cells for electrophoretic mobility shift assays, we demonstrate that S-hexadecyl-CoA was able to increase the mobility of the PPARalpha-containing heterodimer even in the presence of a molar excess of acyl-CoA-binding protein, mimicking the conditions found in vivo.",
keywords = "Acyl Coenzyme A, Acyl-CoA Oxidase, Animals, Cell Line, Chromatography, Affinity, Coenzyme A, DNA-Binding Proteins, Dimerization, Genes, Reporter, Glutathione Transferase, Histone Acetyltransferases, Ligands, Mice, Models, Molecular, Oxidoreductases, Protein Biosynthesis, Protein Conformation, Rats, Receptors, Cytoplasmic and Nuclear, Receptors, Retinoic Acid, Recombinant Proteins, Retinoid X Receptors, Spodoptera, Trans-Activators, Transcription Factors, Transcription, Genetic, Transfection",
author = "M Elholm and I Dam and C Jorgensen and Krogsdam, {A M} and D Holst and I Kratchmarova and M Gottlicher and Gustafsson, {J A} and R Berge and T Flatmark and J Knudsen and S Mandrup and K Kristiansen",
year = "2001",
month = jun,
day = "15",
doi = "10.1074/jbc.M101073200",
language = "English",
volume = "276",
pages = "21410--16",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "Elsevier",
number = "24",
}