TY - JOUR
T1 - Multiple Pit-1-binding sites facilitate estrogen responsiveness of the prolactin gene
AU - Nowakowski, Barbara E.
AU - Maurer, Richard A.
PY - 1994/12
Y1 - 1994/12
N2 - Previous studies have shown that estrogen responsiveness of the rat PRL gene requires the presence of both the estrogen receptor and the tissue- specific transcription factor, Pit-1. To examine the contribution of individual Pit-1-binding sites in permitting an estrogen response, we mutated specific sites in both the proximal and distal regions of the rat PRL gene. The studies reveal that mutation of Pit-1-binding sites in either the proximal or the distal region can have an effect on estrogen responsiveness. The most important Pit-1-binding site appears to be the site in the distal enhancer, which is adjacent to the estrogen receptor-binding site. However, mutation of combinations of other Pit-1-binding sites reveals that these sites also contribute to the estrogen response of the PRL gene. The binding sequences for another transcription factor cannot substitute for Pit-1 sites in bringing about a wild-type estrogen response, as shown by replacement of Pit-1-binding sites with a consensus cAMP-responsive element. Conversion of the imperfect palindromic estrogen response element of the PRL gene to a perfect palindrome eliminated the positive effects of an intact 1D Pit-1- binding site. To examine potential physical interactions between the estrogen receptor and Pit-1, a protein interaction assay was performed. The results demonstrate that labeled estrogen receptor can bind to Pit-1 immobilized on glutathione agarose beads. However, most of the interaction between Pit-1 and the estrogen receptor appears to be DNA dependent. Overall, the results demonstrate a distributed role for multiple Pit-1 sites in permitting an estrogen response of the rat PRL gene.
AB - Previous studies have shown that estrogen responsiveness of the rat PRL gene requires the presence of both the estrogen receptor and the tissue- specific transcription factor, Pit-1. To examine the contribution of individual Pit-1-binding sites in permitting an estrogen response, we mutated specific sites in both the proximal and distal regions of the rat PRL gene. The studies reveal that mutation of Pit-1-binding sites in either the proximal or the distal region can have an effect on estrogen responsiveness. The most important Pit-1-binding site appears to be the site in the distal enhancer, which is adjacent to the estrogen receptor-binding site. However, mutation of combinations of other Pit-1-binding sites reveals that these sites also contribute to the estrogen response of the PRL gene. The binding sequences for another transcription factor cannot substitute for Pit-1 sites in bringing about a wild-type estrogen response, as shown by replacement of Pit-1-binding sites with a consensus cAMP-responsive element. Conversion of the imperfect palindromic estrogen response element of the PRL gene to a perfect palindrome eliminated the positive effects of an intact 1D Pit-1- binding site. To examine potential physical interactions between the estrogen receptor and Pit-1, a protein interaction assay was performed. The results demonstrate that labeled estrogen receptor can bind to Pit-1 immobilized on glutathione agarose beads. However, most of the interaction between Pit-1 and the estrogen receptor appears to be DNA dependent. Overall, the results demonstrate a distributed role for multiple Pit-1 sites in permitting an estrogen response of the rat PRL gene.
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U2 - 10.1210/me.8.12.1742
DO - 10.1210/me.8.12.1742
M3 - Article
C2 - 7708061
AN - SCOPUS:0028580550
SN - 0888-8809
VL - 8
SP - 1742
EP - 1749
JO - Molecular Endocrinology
JF - Molecular Endocrinology
IS - 12
ER -