Interaction of estrogen receptor complexes with the promoter region of genes that are negatively regulated by estrogens: the alpha 2u-globulins.
Since estrogens strongly suppress the expression of alpha 2u-globulin genes in the rat liver, we studied the binding of estrogen-receptor complexes to fragments derived from alpha 2u-globulin gene RAO 01 using a DNA-cellulose competition assay. Rat uterus cytosol labelled with [3H]estradiol was used as a source of the estrogen receptor. As a positive control in these experiments we used an oligonucleotide containing the estrogen response element (ERE) cloned into pUC18. Our experiments indicate that estrogen-receptor complexes bind specifically to the ERE and to a fragment of RAO 01 located in the 5'-upstream region (bp -606 up to -575). This fragment is conserved among other members of this gene family. This is the first time that in vitro estrogen receptor binding is observed to gene fragments derived from a gene that is repressed by this steroid in vivo.
Laboratorium voor Experimentele Geneeskunde en Endocrinologie, Katholieke Universiteit Leuven, Belgium.
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Van Dijck
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Verhoeven
D000510:Alpha-Globulins / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D002482:Cellulose / Q000031:analogs & derivatives; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004958:Estradiol / Q000378:metabolism; D005260:Female; D008969:Molecular Sequence Data; D005810:Multigene Family; D009838:Oligodeoxyribonucleotides; D011401:Promoter Regions, Genetic; D051381:Rats; D011919:Rats, Inbred Strains; D011960:Receptors, Estrogen / Q000302:isolation & purification / Q000378:metabolism*; D015183:Restriction Mapping; D014599:Uterus / Q000378:metabolism / Q000502:physiology*
D000510:Alpha-Globulins; C033190:DNA-cellulose; D009838:Oligodeoxyribonucleotides; D011960:Receptors, Estrogen; C013803:alpha 2u globulin; D004958:Estradiol; D002482:Cellulose; D004247:DNA