Affinity labelling and cloning of steroid and thyroid hormone receptors / edited by H. Gronemeyer.

Date:
[1988]
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Licence: Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)

Credit: Affinity labelling and cloning of steroid and thyroid hormone receptors / edited by H. Gronemeyer. Source: Wellcome Collection.

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    Ch. 16] Analysis of the human glucocorticoid receptor gene promoter 303 extreme 5' end of the human glucocorticoid receptor gene. Hollenberg et al. (1985) isolated human glucocorticoid receptor cDNA clones which contain identical 5' untranslated cDNA sequences. E. Sequence around the human glucocorticoid receptor promoter is G+C rich The nucleotide sequence upstream of the human glucocorticoid receptor RNA start site contains neither a TATA box' nor a 'CAAT box' (Fig. 2). Most other characterized eukaryotic genes have these two sequence elements approximately 30 bp and 80 bp respectively upstream of the RNA initiation site (Breathnach and Chambón, 1981). Since in vitro transcription studies have shown that the 'TATA box' serves to fix the start site of transcription (Fromm and Berg, 1982), it seems surprising that human glucocorticoid receptor gene without a 'TATA box' has a predominant transcriptional start site (Fig. 3). It has been demonstrated that the EGF receptor gene, which possesses similar structural features to the human glucocorticoid receptor gene and has numerous transcriptional initiation sites (Ishii et al., 1985), contains neither a 'TATA box' nor a 'CAAT box'. The sequence between -661 and —1 (Fig. 2), containing the putative promoter, has a G+C content of 72.5%. Further analysis of the human glucocorticoid receptor gene 5' region shows that, similarly to the EGF receptor, a specific sequence CCGCCC and its inverted repeat GGGCGG are present many times (see Fig. 2) in the putative promoter region. The element CCGCCC is the same sequence that is repeated six times within the simian virus 40 (SV40) early promoter (Fromm and Berg, 1982). The 5' region of the human glucocorticoid receptor gene therefore differs from the corresponding regions of most other eukaryotic genes analyzed to date but shares some common features with the SV40 early promoter, the EGF receptor promoter, the mouse metallothionein promoter and the hydroxy methylglutaryl-CoA reduc¬ tase (HMG-CoA reductase) promoter (Ishii et al., 1985; Serfling et al., 1985). The SV40 early promoter contains a GC-rich region, about 80 bp upstream of the RNA start site, which is essential for transcription (Fromm and Berg, 1982; Benoist and Chambón, 1981; Hansen and 5Ьаф, 1983; Wasylyk and Chambón, 1983, Tjian, 1981) and contains six copies of the CCGCCC motif (Ishii et al., 1985 ; Serfling et al., 1985). The gene HMG-CoA reductase does not possess a typical 'TATA box', has five transcriptional start sites (Reynolds et al., 1984) and contains three repeats of the sequence CCGCCC at the 5' flanking region. The c-myc proto-oncogene promoter also contains the CCGCCC motif and is GC rich (see Fig. 6 of Reynolds et al. (1984)). The 5' flanking region of the EGF receptor gene is GC rich and contains five CCGCC repeats. The mouse metallothionein-I gene promoters appear to include two CCGCCC elements present in an inverted configuration (Serfling et al., 1985). Like the corresponding regions of these genes, the 5' flanking region of the human glucocorticoid receptor gene is GC rich. Within this area, there are two repeats of the CCGCCC sequence and three of the inverted elements GGGCGG. Interestingly, the sequence of the human glucocorticoid receptor gene apparently contains in addition a putative glucocorticoid receptor binding site (TGTTCT; see Fig. 2) at position —621. A recent report demonstrates that the human HeLa cell transcription factor Spi