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. 15] The nuclear receptor family: cloning, structure and function 253 E. Functional domains of the glucocorticoid and progesterone receptors 282 1. Glucocorticoid receptor 282 2. Progesterone receptor 283 F. Origin of two chicken progesterone receptors 287 G. How do steroid hormone receptors activate specific gene transcription? 288 Acknowledgments 290 References 290 I. INTRODUCTION The study of biological mechanisms which regulate patterns of gene expression both during the development of eukaryotic organisms and in terminally differentiated cells is one of the major subjects of molecular biology. At the level of transcription this regulation appears to be achieved through positive and negative effects mediated by trans-acting proteins interacting with cis-acting DNA promoter elements (Ptashne, 1986). Steroid hormones trigger complex development and physiological processes. The structural information provided by a given steroid hormone is converted into regulatory transcriptional signals by interaction with specific protein receptors. On binding of the hormone, the receptor gains the ability to alter specifically gene transcription — a process which appears to result from the binding of the hormone-receptor complex to 'enhancer' elements of target gene promoters (Yamamoto, 1985). Thus understanding (1) how the receptor protein is able to specifically recognize its cognate hormone, (2) how this binding results in the interaction of the receptor with target gene promoter elements, (3) how this interaction leads to initiation of specific gene transcription, and (4) what the role of the hormone is in these processes, are prerequisites to understanding the molecular mechanism of steroid-hormone-controlled gene activity. Such an analysis is now possible as a result of the cloning of cDNAs corresponding to the estrogen (Walter et al., 1985; Krust et al., 1986; Koike et al., 1987; Weiler et al., 1987; White et al., 1987), glucocorticoid (Miesfeld et al., 1984; Weinberger et al., 1985a,b; Govindan et al., 1985; Danielson et al., 1986), progesterone (Jeltsch et al., 1986; Conneely et al., 1986; Lx)0sfelt et al., 1986; Misrahi et al., 1987), and vitamin D (McDonnell et al., 1987) receptors. We review here the results obtained recently, with a particular emphasis on the work performed in our laboratory. П. cDNA CLONING A. Human estrogen receptor Two independent approaches were used to isolate cDNAs corresponding to the estrogen receptor (Walter et al., 1985) from the human breast cancer cell Une MCF- 7. PolyA* RNA was fractionated on sucrose gradients and assayed for the presence of receptor mRNA by in vitro translation and immunoprecipitation with monoclonal antibodies directed against the human estrogen receptor. Fractions enriched in receptor mRNA were pooled and used for randomly primed cDNA synthesis and cloning into Xgt vectors. The >.gtll-cDNA expression library was then screened with a panel of monoclonal anti-estrogen receptor antibodies. Out of more than 1000000