The orphan nuclear receptor TR2 suppresses a DR4 hormone response element of the mouse CRABP-I gene promoter.
The mouse orphan nuclear receptor TR2-11-f suppressed the expression of reporters fused to a hormone response element of the mouse cellular retinoic acid-binding protein I gene promoter. TR2-11-f was able to bind to a direct repeat with four nucleotides in the spacer (5'TGACCTTTGGGGACCT3') located within this hormone response element as homodimers. The specificity of protein-DNA interactions was demonstrated by competition in gel retardation and antibody-mediated supershift reactions. The residues critical for TR2-11-f binding were mapped to both repeated sequences, whereas the spacer and the flanking sequences were less important. The Kd and Bmax of TR2-11-f homodimer binding to this direct repeat were determined to be 2.6 nM and 0.012 nM, respectively. By using a yeast two-hybrid system, it was demonstrated that dimerization of TR2-11-f was mediated by its ligand-binding domain. The actions of TR2-11-f in regulating cellular retinoic acid-binding protein I gene will likely influence retinoic action and availability within the cells.
Chinpaisal
C
C
Department of Pharmacology, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA.
Chang
L
L
Hu
X
X
Lee
C H
CH
Wen
W N
WN
Wei
L N
LN
D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D019556:COS Cells; D004268:DNA-Binding Proteins / Q000378:metabolism*; D019281:Dimerization; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D057107:Nuclear Receptor Subfamily 2, Group C, Member 1; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D018168:Receptors, Retinoic Acid / Q000096:biosynthesis / Q000235:genetics*; D011988:Receptors, Thyroid Hormone / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013489:Suppression, Genetic; D014212:Tretinoin / Q000378:metabolism
D004268:DNA-Binding Proteins; D008024:Ligands; C539823:Nr2c1 protein, mouse; D009687:Nuclear Proteins; D057107:Nuclear Receptor Subfamily 2, Group C, Member 1; D018168:Receptors, Retinoic Acid; D011988:Receptors, Thyroid Hormone; D011994:Recombinant Proteins; C082927:retinoic acid binding protein I, cellular; D014212:Tretinoin