Binding of the Human Glucocorticoid Receptor to Defined Regions in the Human Growth Hormone and Placental Lactogen Genes

Eliard, P H; Marchand, Maurice; Rousseau, Guy; Formstecher, Pierre; Mathy-Hartert, M.; Belayew, Alexandra; Martial, Joseph

doi:10.1089/dna.1985.4.409

Cited by 28 publications

(12 citation statements)

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“…The cloning of the GH-N and CS-B genes and the purification and preparation of blunt-ended fragments have been described (12). The DNA-cellulose-binding assay for thyroid hormone receptors was designed on the basis of the competition assay used to study the binding of unpurified gluco-…”

Section: Methodsmentioning

confidence: 99%

“…The cloning of the GH-N and CS-B genes and the purification and preparation of blunt-ended fragments have been described (12 (Fig. 1) was as described for other mammalian thyroid hormone receptors (2).…”

Section: Methodsmentioning

confidence: 99%

“…Cell Biology: Barlow et (12). This downstream region contains, in the first intron of the gene, a 15-bp consensus sequence that is found in a number of glucocorticoid-responsive genes (10)(11)(12).…”

Section: Methodsmentioning

confidence: 99%

“…Thyroid hormone receptor content and affinity were assessed by incubating 38. of an ice-cold solution of hydroxyapatite (15 g/100 ml in 20 mM Tris HCl/10 mM KH2PO4, pH 7.2) was added to the tubes and the mixture was further incubated for 10 min on ice. After centrifugation (1000 x g for 2 min), the hydroxyapatite pellet was washed three times with 10 mM Tris HCl, 5 mM NaH2PO4, 1.5 mM EDTA, and 0.5% Triton X-100 (pH 7.2), and the residual radioactivity was determined.DNA-Binding Assay for the Thyroid Hormone Receptor.The cloning of the GH-N and CS-B genes and the purification and preparation of blunt-ended fragments have been described (12). The DNA-cellulose-binding assay for thyroid hormone receptors was designed on the basis of the competition assay used to study the binding of unpurified gluco-…”

mentioning

confidence: 99%

“…Binding regions for the rat (10,11) and human (12) glucocorticoid receptors have been identified in the human GH gene upstream and downstream from the transcription initiation site (cap site). Homologous binding regions have also been detected (12) in the human placental lactogen (chorionic somatomammotropin, CS) gene, which has a 92% overall nucleotide sequence homology with the human GH gene (13). In contrast, the question of specific recognition sites for thyroid hormone receptors on any DNA has remained open.…”

mentioning

confidence: 99%

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Thyroid hormone receptors bind to defined regions of the growth hormone and placental lactogen genes.

Barlow¹,

Voz²,

Eliard³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

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The intracellular receptor for thyroid hormone is a protein found in chromatin. Since thyroid hormone stimulates transcription of the growth hormone gene through an unknown mechanism, the hypothesis that the thyroid hormone-receptor complex interacts with defined regions of this gene has been investigated in a cell-free system. Nuclear extracts from human lymphoblastoid IM-9 cells containing thyroid hormone receptors were incubated with L-3,5,3'-tri[('I]iodothyronine and calf thymus DNA-cellulose. Restriction fragments of the human growth hormone gene were added to determine their ability to inhibit labeled receptor binding to DNA-cellulose. These fragments encompassed nucleotide sequences from about three kilobase pairs upstream to about four kilobase pairs downstream from the transcription initiation site. The thyroid hormone-receptor complex bound preferentially to the 5'-flanking sequences of the growth hormone gene in a region between nucleotide coordinates -290 and -129. The receptor also bound to an analogous promoter region in the human placental lactogen gene, which has 92% nucleotide sequence homology with the growth hormone gene. These binding regions appear to be distinct from those that are recognized by the receptor for glucocorticoids, which stimulate growth hormone gene expression synergistically with thyroid hormone. The presence of thyroid hormone was required for binding of its receptor to the growth hormone gene promoter, suggesting that thyroid hormone renders the receptor capable of recognizing specific gene regions. L-3,5,3'-Triiodothyronine (T3), the active form of thyroid hormone, modulates the expression of a number of genes (1, 2), and this is ascribed to the association of T3 with receptors localized in chromatin (3). In rat pituitary tumor cells the concentration of growth hormone (GH) mRNA increases in response to physiological concentrations ofT3 (refs. 4 and 5).Although additional mechanisms have not been ruled out, this response reflects an increased rate of transcription of the GH gene (6, 7), presumably through a direct effect of the receptor (8). Moreover, T3 acts synergistically with glucocorticoid hormones in stimulating GH gene transcription (6, 9). Binding regions for the rat (10, 11) and human (12) glucocorticoid receptors have been identified in the human GH gene upstream and downstream from the transcription initiation site (cap site). Homologous binding regions have also been detected (12) in the human placental lactogen (chorionic somatomammotropin, CS) gene, which has a 92% overall nucleotide sequence homology with the human GH gene (13). In contrast, the question of specific recognition sites for thyroid hormone receptors on any DNA has remained open. We have now searched for such sites in the human GH and CS genes (12) using the human thyroid hormone receptor in a cell-free system. This system contained [1251]T3-labeled nuclear extracts from cultured human lymphoblastoid cells of the IM-9 line and cloned fragments from the genes of interest. MATERIALS AND M...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%