The region of CQQQKPQRRP of PGC-1alpha interacts with the DNA-binding complex of FXR/RXRalpha.
PGC-1alpha co-activates transcription by several nuclear receptors. To study the interaction among PGC-1alpha, RXRalpha/FXR, and DNA, we performed electrophoresis mobility shift assays. The RXRalpha/FXR proteins specifically bound to DNA containing the IR-1 sequence in the absence of ligand. When the fusion protein of GST-PGC-1alpha was added to the mixture of RXRalpha/FXR/DNA, the ligand-influenced retardation of the mobility was observed. The ligand for RXRalpha (9-cis-retinoic acid) was necessary for this retardation, whereas, the ligand for FXR, chenodeoxycholic acid, barely had an effect. The results obtained using truncated PGC-1alpha proteins suggested that two regions are necessary for PGC-1alpha to interact with the DNA-binding complex of RXRalpha/FXR. One is the region of the second leucine-rich motif, and the other is that of the amino acid sequence CQQQKPQRRP, present between the second and third leucine-rich motifs. The results obtained with the SPQSS mutation for KPQRR suggested that the basic amino acids are important for the interaction.
Department of Molecular Biology, Biomolecular Engineering Research Institute (BERI), Osaka, Japan.
Eiko
E
Kanaya
Hisato
H
Jingami
D000595:Amino Acid Sequence; D001667:Binding, Competitive; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D006360:Heat-Shock Proteins / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism*; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; D011485:Protein Binding; D018160:Receptors, Cytoplasmic and Nuclear; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D047490:Retinoid X Receptor alpha / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D000099150:Receptor, Farnesoid X-Activated
D004247:DNA; D004268:DNA-Binding Proteins; D006360:Heat-Shock Proteins; D010455:Peptides; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; D018160:Receptors, Cytoplasmic and Nuclear; D011993:Recombinant Fusion Proteins; D047490:Retinoid X Receptor alpha; D014157:Transcription Factors; D000099150:Receptor, Farnesoid X-Activated; C486497:PPARGC1A protein, human