Identification of human TR2 orphan receptor response element in the transcriptional initiation site of the simian virus 40 major late promoter.
A DNA response element (TR2RE-SV40) for the TR2 orphan receptor, a member of the steroid-thyroid hormone receptor superfamily, has been identified in the simian virus 40 (SV40) +55 region (nucleotide numbers 368-389, 5'-GTTAAGGTTCGTAGGTCATGGA-3'). Electrophoretic mobility shift assay, using in vitro translated TR2 orphan receptor with a molecular mass of 67 kilodaltons, showed a specific binding with high affinity (dissociation constant = 9 nM) for this DNA sequence. DNA-swap experiments using chloramphenicol acetyl-transferase assay demonstrated that androgen can suppress the transcriptional activities of SV40 early promoter via the interaction between this TR2RE-SV40 and the chimeric receptor AR/TR2/AR with the DNA-binding domain of the TR2 orphan receptor flanked by the N-terminal and androgen-binding domains of the androgen receptor. In addition, this TR2RE-SV40 can function as a repressor to suppress the transcriptional activities of both SV40 early and late promoters. Together, these data suggest the TR2RE-SV40 may represent the first identified natural DNA response element for the TR2 orphan receptor that may function as a repressor for the SV40 gene expression.
Lee
H J
HJ
Department of Human Oncology, University of Wisconsin, Madison 53792.
Chang
C
C
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D057107:Nuclear Receptor Subfamily 2, Group C, Member 1; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D011817:Rabbits; D011988:Receptors, Thyroid Hormone / Q000096:biosynthesis / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D012156:Reticulocytes / Q000378:metabolism; D013539:Simian virus 40 / Q000235:genetics*; D013963:Thyroid Hormones / Q000378:metabolism*; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; C539822:NR2C1 protein, human; D057107:Nuclear Receptor Subfamily 2, Group C, Member 1; D011988:Receptors, Thyroid Hormone; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D013963:Thyroid Hormones; D015500:Chloramphenicol O-Acetyltransferase