Ligand-regulated heterodimerization of peroxisome proliferator-activated receptor α with liver X receptor α.
Peroxisome proliferator-activated receptor α (PPARα) and liver X receptor α (LXRα) are members of the nuclear receptor superfamily that function to regulate lipid metabolism. Complex interactions between the LXRα and PPARα pathways exist, including competition for the same heterodimeric partner, retinoid X receptor α (RXRα). Although data have suggested that PPARα and LXRα may interact directly, the role of endogenous ligands in such interactions has not been investigated. Using in vitro protein-protein binding assays, circular dichroism, and co-immunoprecipitation of endogenous proteins, we established that full-length human PPARα and LXRα interact with high affinity, resulting in altered protein conformations. We demonstrated for the first time that the affinity of this interaction and the resulting conformational changes could be altered by endogenous PPARα ligands, namely long chain fatty acids (LCFA) or their coenzyme A thioesters. This heterodimer pair was capable of binding to PPARα and LXRα response elements (PPRE and LXRE, respectively), albeit with an affinity lower than that of the respective heterodimers formed with RXRα. LCFA had little effect on binding to the PPRE but suppressed binding to the LXRE. Ectopic expression of PPARα and LXRα in mammalian cells yielded an increased level of PPRE transactivation compared to overexpression of PPARα alone and was largely unaffected by LCFA. Overexpression of both receptors also resulted in transactivation from an LXRE, with decreased levels compared to that of LXRα overexpression alone, and LCFA suppressed transactivation from the LXRE. These data are consistent with the hypothesis that ligand binding regulates heterodimer choice and downstream gene regulation by these nuclear receptors.
Department of Biochemistry and Molecular Biology, Boonshoft School of Medicine, Wright State University , Dayton, Ohio 45435, United States.
Madhumitha
M
Balanarasimha
Andrea M
AM
Davis
Frances L
FL
Soman
S Dean
SD
Rider
Heather A
HA
Hostetler
D002942:Circular Dichroism; D003065:Coenzyme A / Q000737:chemistry / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D047468:Immunoprecipitation; D008024:Ligands; D000071518:Liver X Receptors; D057093:Orphan Nuclear Receptors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D047493:PPAR alpha / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010171:Palmitoyl Coenzyme A / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D055503:Protein Multimerization; D020218:Response Elements
D005227:Fatty Acids; D008024:Ligands; D000071518:Liver X Receptors; C000605213:NR1H3 protein, human; D057093:Orphan Nuclear Receptors; D047493:PPAR alpha; D010171:Palmitoyl Coenzyme A; D003065:Coenzyme A