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We have defined and characterized a region upstream of the bovine prolactin gene that confers repression by glucocorticoids. This 'negative glucocorticoid response element' (nGRE) contains multiple footprinting sites for purified glucocorticoid receptor protein between -51 and -562 bp. A strong consensus sequence for receptor binding within the nGRE has not yet been defined, but it is apparent that nGRE sequences differ from the GRE consensus elements that confer positive glucocorticoid regulation. Unlike 'positive' GREs, the nGRE enhances promoter activity in the absence of glucocorticoids or receptor, presumably through the action of a protein that binds in the same region and activates transcription. The hormone-receptor complex appears to negate this enhancement by competing or inactivating the second factor. As with positive GREs, nGRE sequences confer hormonal regulation upon linked heterologous promoters within various cell types; a 34-bp subfragment containing a single receptor binding site is sufficient for nGRE activity. We speculate that nGRE sequences might alter the structure of bound receptor, thereby preventing it from functioning as a positive regulator when bound at those sites.
Sakai et al. (Thu,) studied this question.