DNA-binding properties of the overexpressed recombinant estrogen receptor alpha.
Estrogen receptor alpha was overexpressed in COS-7 cells by transformation of an expression vector with the full length open reading frame of the receptor alpha. The nuclear estradiol-receptor complex and the soluble receptor from the COS-7 cells were cross-linked with an estrogen response element (ERE), which was substituted with 5-bromo-2'-deoxyuridine (BrdUVRE), as the receptor dimers by UV irradiation. In gel retardation analysis, the specific bindings of both nuclear and soluble receptors to ERE were decreased with increasing of KCl concentration compared with 0.1 M KCl. The ionic interactions of both receptors with ERE are thought to be similar.
Department of Biochemistry Kawasaki Medical School, Okayama, Japan.
M
M
Ikeda
N
N
Tsuji
K
K
Kikukawa
Y
Y
Asahara
A
A
Nakashima
Y
Y
Minatogawa
D000818:Animals; D000911:Antibodies, Monoclonal; D001973:Bromodeoxyuridine; D019556:COS Cells; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D019281:Dimerization; D004958:Estradiol / Q000378:metabolism; D047628:Estrogen Receptor alpha; D015870:Gene Expression; D011189:Potassium Chloride / Q000494:pharmacology; D011960:Receptors, Estrogen / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D012995:Solubility; D014466:Ultraviolet Rays
D000911:Antibodies, Monoclonal; D047628:Estrogen Receptor alpha; D011960:Receptors, Estrogen; D011994:Recombinant Proteins; D004958:Estradiol; D011189:Potassium Chloride; D004247:DNA; D001973:Bromodeoxyuridine