Molecular Model of the Interaction Between the Glucocorticoid Receptor and the Regulatory Elements of Inducible Genes

Scheidereit, Claus; Westphal, Hannes M.; Carlson, Christopher; Bosshard, Heinz E.; Beato, Miguel

doi:10.1089/dna.1986.5.383

Cited by 115 publications

(31 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…CAANNTGTTCT-3', with the 3' hexamer stringently conserved (35). The promoter region of the rat AFP gene Ap.…”

Section: Resultsmentioning

confidence: 99%

“…Considering the enhancer properties of inducible glucocorticoid response elements (7,25), one may further speculate that the AFP glucocorticoid response element might be a glucocorticoid receptor-repressible enhancer. It has also been proposed that glucocorticoid receptors induce and repress genes by using different functional domains (29); perhaps significantly, the left and right segments of the consensus-inducible glucocorticoid response element (35) …”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Enhancer and promoter elements directing activation and glucocorticoid repression of the alpha 1-fetoprotein gene in hepatocytes.

Guertin¹,

LaRue²,

Bernier³

et al. 1988

Mol. Cell. Biol.

109

View full text Add to dashboard Cite

Mutations were introduced in 7 kilobases of 5'-flanking rat ac-fetoprotein (AFP) genomic DNA, linked to the chloramphenicol acetyltransferase gene. AFP promoter activity and its repression by a glucocorticoid hormone were assessed by stable and transient expression assays. Stable transfection assays were more sensitive and accurate than transient expression assays in a Morris 7777 rat hepatoma recipient (Hepa7.6), selected for its strong AFP repression by dexamethasone. The segment of DNA encompassing a hepatocyte-constitutive chromatin DNase I-hypersensitive site at -3.7 kilobases and a liver developmental stage-specific site at -2.5 kilobases contains interacting enhancer elements sufficient for high AFP promoter activity in Hepa7.6 or HepG2 cells. Deletions and point mutations define an upstream promoter domain of AFP gene activation, operating with at least three distinct promoter-activating elements, PEI at -65 base pairs, PEII at -120 base pairs, and DE at -160 base pairs. PEI and PEII share homologies with albumin promoter sequences, PEII is a near-consensus nuclear factor I recognition sequence, and DE overlaps a glucocorticoid receptor recognition sequence. An element conferring glucocorticoid repression of AFP gene activity is located in the upstream AFP promoter domain. Receptor-binding assays indicate that this element is the glucocorticoid receptor recognition sequence which overlaps with promoter-activating element DE.The a,-fetoprotein (AFP) gene, a member of the albumin gene family, is expressed by fetal or malignant hepatocytes and repressed in normal mature hepatocytes. This forms the basis of a long-exploited model system to study cell differentiation and its impairment in neoplasia (1). Molecular genetics have brought the AFP model to molecular levels of cell differentiation, toward finely discerning how genes respond to or escape from developmental and growth signals (2,40,41). As yet, little is known about how differential regulation is exerted on the tandemly organized AFPalbumin locus and whether the two genes operate under shared elements of control. However, at least one wellcharacterized transcription factor, the glucocorticoid receptor, is known to selectively shut off the AFP gene in the developing liver (4, 21). This hormonal effect reaches into the mechanisms of neoplastic resistance to differentiation, because the AFP gene in malignant cells is generally refractory to glucocorticoids (2).Analyses of rat AFP gene and chromatin structures have identified domains in the 5' region of the locus potentially involved in its liver-specific, developmental stage-dependent, and glucocorticoid-regulated expression. A promoter domain spanning '230 nucleotides comprises a chromatin DNase I-hypersensitive (DH) site (32, 42, 44) that is selectively suppressed by dexamethasone (44), glucocorticoid receptor recognition sequences, and octamer motifs similar to those present in other genes under developmental and growth control (8). A distal domain (-2 to -4 kilobases [kb] relative to the AFP trans...

show abstract

“…CAANNTGTTCT-3', with the 3' hexamer stringently conserved (35). The promoter region of the rat AFP gene Ap.…”

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Enhancer and promoter elements directing activation and glucocorticoid repression of the alpha 1-fetoprotein gene in hepatocytes.

Guertin¹,

LaRue²,

Bernier³

et al. 1988

Mol. Cell. Biol.

109

View full text Add to dashboard Cite

show abstract

“…The absence of any effect by the other steroids-in particu- induction. To obtain additional support for this surprising observation, the 15-bp oligonucleotide was modified at several sites, some of which are known to be contacted by the glucocorticoid receptor (19,20). The effect of these single or double substitutions upon inducibility by dexamethasone and R 5020 was tested as before.…”

Section: Resultsmentioning

confidence: 99%

A DNA sequence of 15 base pairs is sufficient to mediate both glucocorticoid and progesterone induction of gene expression.

Strähle

Klöck

Schütz

1987

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

311

126

View full text Add to dashboard Cite

To defmie the recognition sequence of the glucocorticoid receptor and its relationship with that of the progesterone receptor, oligonucleotides derived from the glucocorticoid response element of the tyrosine aminotransferase gene were tested upstream of a heterologous promoter for their capacity to mediate effects of these two steroids. We show that a 15-base-pair sequence with partial symmetry is sufficient to confer glucocorticoid inducibility on the promoter of the herpes simplex virus thymidine kinase gene. The same 15-base-pair sequence mediates induction by progesterone. Point mutations in the recognition sequence affect inducibility by glucocorticoids and progesterone similarly. Together with the strong conservation of the sequence of the DNA-binding domain of the two receptors, these data suggest that both proteins recognize a sequence that is similar, if not the same.The effect of steroid hormones on specific gene transcription is mediated by receptors to which the hormones bind with high selectivity and affinity. Activation of transcription results from binding of the hormone-receptor complex to specific sequences of inducible genes (1, 2). Surprisingly, it has been observed that fragments of the chicken lysozyme gene and of the long terminal repeat of mouse mammary tumor virus that confer glucocorticoid inducibility also allow progesterone induction (3, 4). The glucocorticoid and progesterone receptors bind to similar sequences within these fragments, suggesting that the binding sites may be closely related, but distinct (5, 6). Both types of receptors also bind to partially overlapping sequences in the rabbit uteroglobin gene (7,8). These sequences, however, have not been functionally characterized. To define the sequence requirements for specific binding of the glucocorticoid receptor and to determine the degree to which this sequence overlaps with the recognition sequence for the progesterone receptor, a minimal glucocorticoid response element (GRE) 15 base pairs (bp) in length was defined. This 15-bp sequence is part of the GRE region of the tyrosine aminotransferase (TAT) gene protected against DNase I digestion by the purified glucocorticoid receptor (9). We demonstrate here that the same 15-mer which confers responsiveness to glucocorticoids also mediates progesterone induction. Since point mutations in this 15-bp sequence affect inducibility by both steroids similarly, we suggest that the recognition sequences of the glucocorticoid and progesterone receptors are similar, if not identical. MATERIALS AND METHODSPlasmid Constructions. Oligonucleotides were prepared with an Applied Biosystems 380A DNA synthesizer and purified as recommended by the manufacturer. Annealing of the complementary strands, which generated BamHI or Xba I cohesive ends, and cloning upstream of the herpes simplex virus thymidine kinase (TK) promoter in pBLCAT2 (10) were done by standard procedures (11). pBLCAT2 contains the complete TK promoter (-105 to +62) linked to the chloramphenicol acetyltransferase (CAT) co...

show abstract

“…Close to the center of symmetry of the ERED is the conserved hexanucleotide 5'-TGTTCT-3' on the lower (or antisense strand), which is the core motif for binding to DNA by the glucocorticoid receptor. In vitro receptor-DNA binding studies have shown that this hexanucleotide and neighboring sequences on the VTG II gene are bound by the glucocorticoid receptor (26) and could also possibly be recognized by the androgen receptor (6). The hexanucleotide motif is also part of the recognition sequence of the progesterone receptor (1,14,29).…”

mentioning

confidence: 99%

“…The boxed sequence is the conserved hexanucleotide motif 5'-TGTTCT-3' (in the reversed orientation) shown to bind the glucocorticoid receptor (26). tk = TK.…”

mentioning

confidence: 99%

Estrogen and progesterone receptor-binding sites on the chicken vitellogenin II gene: synergism of steroid hormone action.

Cato¹,

Heitlinger²,

Ponta³

et al. 1988

Mol. Cell. Biol.

View full text Add to dashboard Cite

The chicken vitellogenin II gene is transcriptionally activated by estrogens. In transient transfection experiments in human T47D cells that contain receptors for various steroids, we showed estradiol, progestin, and androgen responses of a chimeric chicken vitellogenin II construct. This construct consists of DNA sequences from -626 to -590 upstream of the start of transcription of the chicken viteliogenin gene linked to the herpes simplex virus thymidine kinase promoter driving the transcription of the bacterial chloramphenicol acetyltransferase gene. Treatment of the transfected T47D cells with a combination of estradiol and the progestin R5020 led to a superinduction of chloramphenicol acetyltransferase activity, showing a synergistic action of these two steroids. This synergism was not observed upon treatment of the transfected cells with estradiol and the androgen dihydrotestosterone. Using point mutations in the vitellogenin gene fragment, we showed in functional and in in vitro DNase I footprinting assays with a purified progesterone receptor that, for the synergistic action of estradiol and R5020 to occur, the progesterone receptor must be bound to the vitellogenin gene fragment. The progesterone receptor-binding site was localized at -610 to -590, close to the consensus sequence (-626 to -613) for estrogen receptor binding and function. We therefore demonstrate here that two different steroid hormones can be functionally synergistic through the interaction of their corresponding receptors with two different binding sites adjacent to one another.The regulation of gene expression by steroid hormones is thought to occur through the interaction of steroid hormone receptor complexes with discrete nucleotide sequences in the neighborhood of the promoters of inducible genes (for reviews, see references 2 and 35). Short perfect or imperfect palindromic sequences that mediate steroid hormone action have been identified near the promoters of a number of steroid hormone-regulated genes. For example, 13-base-pair (bp) palindromic elements that mediate the estrogen response have been identified near the promoters of the Xenopus and chicken vitellogenin genes (4,17,18,19,22). Similarly, 15-bp perfect or imperfect palindromes that encompass the hexanucleotide motif 5'-TGTT/CCT-3' have been shown to mediate glucocorticoid and progestin responses in a number of genes (5,6,29,30). The delimitation of these short nucleotide sequences was achieved through gene transfer studies with either in vitro mutagenesis of known hormone receptor-binding sites (6) or different chimeras containing synthetic oligonucleotides (13,17,30). In a number of hormone response elements, such as that in mouse mammary tumor virus DNA (3, 6), the tyrosine aminotransferase gene (13), and the Xenopus vitellogenin Bi and chicken vitellogenin II (VTG II) genes (4,16,22,27) sites function by cooperating with the binding sites for other transcriptional factors that lie in their immediate vicinity.In the VTG II gene, a gene whose transcription is known t...

show abstract

Molecular Model of the Interaction Between the Glucocorticoid Receptor and the Regulatory Elements of Inducible Genes

Cited by 115 publications

References 24 publications

Enhancer and promoter elements directing activation and glucocorticoid repression of the alpha 1-fetoprotein gene in hepatocytes.

Enhancer and promoter elements directing activation and glucocorticoid repression of the alpha 1-fetoprotein gene in hepatocytes.

A DNA sequence of 15 base pairs is sufficient to mediate both glucocorticoid and progesterone induction of gene expression.

Estrogen and progesterone receptor-binding sites on the chicken vitellogenin II gene: synergism of steroid hormone action.

Contact Info

Product

Resources

About