Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Binding, Competitive
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DNA / metabolism*
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / isolation & purification
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DNA-Binding Proteins / metabolism*
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Electrophoretic Mobility Shift Assay
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Heat-Shock Proteins / chemistry*
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Heat-Shock Proteins / metabolism*
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Humans
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Molecular Sequence Data
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Peptides / metabolism*
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Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha
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Protein Binding
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Receptors, Cytoplasmic and Nuclear
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Recombinant Fusion Proteins / isolation & purification
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Recombinant Fusion Proteins / metabolism
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Retinoid X Receptor alpha / chemistry
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Retinoid X Receptor alpha / isolation & purification
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Retinoid X Receptor alpha / metabolism*
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Transcription Factors / chemistry*
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Transcription Factors / isolation & purification
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Transcription Factors / metabolism*
Substances
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DNA-Binding Proteins
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Heat-Shock Proteins
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PPARGC1A protein, human
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Peptides
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Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha
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Receptors, Cytoplasmic and Nuclear
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Recombinant Fusion Proteins
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Retinoid X Receptor alpha
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Transcription Factors
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farnesoid X-activated receptor
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DNA