Fatty acids and hypolipidemic drugs regulate peroxisome proliferator-activated receptors alpha - and gamma-mediated gene expression via liver fatty acid binding protein: a signaling path to the nucleus.

Peroxisome proliferator-activated receptor alpha (PPARalpha) is a key regulator of lipid homeostasis in hepatocytes and target for fatty acids and hypolipidemic drugs. How these signaling molecules reach the nuclear receptor is not known; however, similarities in ligand specificity suggest the liver fatty acid binding protein (L-FABP) as a possible candidate. In localization studies using laser-scanning microscopy, we show that L-FABP and PPARalpha colocalize in the nucleus of mouse primary hepatocytes. Furthermore, we demonstrate by pull-down assay and immunocoprecipitation that L-FABP interacts directly with PPARalpha. In a cell biological approach with the aid of a mammalian two-hybrid system, we provide evidence that L-FABP interacts with PPARalpha and PPARgamma but not with PPARbeta and retinoid X receptor-alpha by protein-protein contacts. In addition, we demonstrate that the observed interaction of both proteins is independent of ligand binding. Final and quantitative proof for L-FABP mediation was obtained in transactivation assays upon incubation of transiently and stably transfected HepG2 cells with saturated, monounsaturated, and polyunsaturated fatty acids as well as with hypolipidemic drugs. With all ligands applied, we observed strict correlation of PPARalpha and PPARgamma transactivation with intracellular concentrations of L-FABP. This correlation constitutes a nucleus-directed signaling by fatty acids and hypolipidemic drugs where L-FABP acts as a cytosolic gateway for these PPARalpha and PPARgamma agonists. Thus, L-FABP and the respective PPARs could serve as targets for nutrients and drugs to affect expression of PPAR-sensitive genes.

Molecular diversity of plaques of epithelial-adhering junctions.

In biochemical and immunocytochemical comparisons of adhering junctions of different epithelia, we have observed differences in molecular composition not only between the intermediate filament-attached desmosomes and the actin filaments-anchoring adherens junctions but also between desmosomes of different tissues and of different strata in the same stratified epithelium. In addition we now report cell type-specific differences of molecular composition and immunoreactivity in both desmosomes and adherens junctions of certain simple epithelia. Whereas the zonula adhaerens of human intestinal and colonic epithelial cells, and of carcinomas derived therefrom, contains the additional armadillo-type plaque protein ARVCF, this protein has not been detected in the zonula adhaerens of hepatocytes. Similarly, plakophilin 3 is present in the desmosomal plaques of intestinal and colonic cells but appears to be absent from the hepatocytic desmosomes. We suggest that these profound compositional differences in the junctions of related simple epithelia are correlated to functional differences of the specific type of epithelium.