Identification of the human aldolase A gene as the first induced target for the TR2 orphan receptor, a member of the steroid hormone receptor superfamily.
The human TR2 orphan receptor (TR2) is a member of the steroid/thyroid hormone receptor superfamily that regulates the transcription of complex gene networks and subsequently controls diverse aspects of growth, development, and differentiation. In the present study, we have found that the TR2 is one of the M1 site (nucleotide numbers 2017-2034, 5'-AAAAGGGCAGGGGTCATT-3') binding proteins of the muscle-specific pM promoter in the human aldolase A gene. Electrophoretic mobility shift assay (EMSA) showed a specific binding with high affinity (dissociation constant = 4.6 nM) between the TR2 and the M1 element. Circular permutation assay revealed a localized DNA flexibility induced by the TR2 binding, and the bend angle was estimated to be 73 +/- 2 degrees. Furthermore, a dual-luciferase reporter gene assay demonstrated that the TR2 may enhance the expression of luciferase activities via the wild-type M1 site but not the mutant M1 element in human QM7 muscle myoblasts. In conclusion, our data represent the first case of demonstrating that the TR2 may serve as a transcriptional inducer in muscle-specific aldolase A gene expression.
Chang
C
C
University of Wisconsin Comprehensive Cancer Center, Madison 53792, USA.
Lee
H J
HJ
Lee
Y F
YF
D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D015342:DNA Probes; D004268:DNA-Binding Proteins / Q000378:metabolism; D019281:Dimerization; D005634:Fructose-Bisphosphate Aldolase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology; D057107:Nuclear Receptor Subfamily 2, Group C, Member 1; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011987:Receptors, Steroid / Q000737:chemistry / Q000378:metabolism*; D011988:Receptors, Thyroid Hormone / Q000737:chemistry / Q000378:metabolism*; D014162:Transfection / Q000235:genetics
D015342:DNA Probes; D004268:DNA-Binding Proteins; C539822:NR2C1 protein, human; D057107:Nuclear Receptor Subfamily 2, Group C, Member 1; D011987:Receptors, Steroid; D011988:Receptors, Thyroid Hormone; D008156:Luciferases; D005634:Fructose-Bisphosphate Aldolase