TY - JOUR
T1 - Effects of estradiol and 4-hydroxytamoxifen on the conformation, thermal stability, and DNA recognition of estrogen receptor β
AU - Vijayanathan, Veena
AU - Greenfield, Norma J.
AU - Thomas, T. J.
AU - Ivanova, Margarita M.
AU - Tyulmenkov, Valentyn V.
AU - Klinge, Carolyn M.
AU - Gallo, Michael A.
AU - Thomas, Thresia
PY - 2007/2
Y1 - 2007/2
N2 - Estrogen receptors (ERα and ERβ) are ligand-activated transcription factors. We examined the effects of estradiol (E2), 4-hydroxytamoxifen (HT), and the estrogen response element (ERE) on the helical content and thermal unfolding of ERβ. A circular dichroism (CD) spectrum of ERβ showed changes at 210 and 222 nm that were due to the presence of E2, which is indicative of partial unfolding. In contrast, HT did not alter the CD spectrum of ERβ. The addition of E2 + ERE caused an increase in the α-helical content and an increase in the temperature midpoint of folding transition (TM) from 39 ± 0.7°C to 57.2 ± 1°C. The addition of E2 + mutant ERE, or E 2 + control oligonucleotide, increased the TM of ERβ to 45 ± 2°C only. In the presence of HT, ERβ yielded similar TM values (55-58°C) with ERE, mutant ERE, or control oligodeoxynucleotide. The binding affinity of ERβ for ERE increased 125.7-fold as a result of the presence of E2, but only 4-fold as a result of HT. These results demonstrate coupled effects of E2 and ERE on ERβ stability and binding affinity. The increased thermal stability of HT-ERβ-ERE was associated with reduced specificity of ERβ-ERE recognition, illustrating profound differences in conformational states of ERβ induced by E2 and HT.
AB - Estrogen receptors (ERα and ERβ) are ligand-activated transcription factors. We examined the effects of estradiol (E2), 4-hydroxytamoxifen (HT), and the estrogen response element (ERE) on the helical content and thermal unfolding of ERβ. A circular dichroism (CD) spectrum of ERβ showed changes at 210 and 222 nm that were due to the presence of E2, which is indicative of partial unfolding. In contrast, HT did not alter the CD spectrum of ERβ. The addition of E2 + ERE caused an increase in the α-helical content and an increase in the temperature midpoint of folding transition (TM) from 39 ± 0.7°C to 57.2 ± 1°C. The addition of E2 + mutant ERE, or E 2 + control oligonucleotide, increased the TM of ERβ to 45 ± 2°C only. In the presence of HT, ERβ yielded similar TM values (55-58°C) with ERE, mutant ERE, or control oligodeoxynucleotide. The binding affinity of ERβ for ERE increased 125.7-fold as a result of the presence of E2, but only 4-fold as a result of HT. These results demonstrate coupled effects of E2 and ERE on ERβ stability and binding affinity. The increased thermal stability of HT-ERβ-ERE was associated with reduced specificity of ERβ-ERE recognition, illustrating profound differences in conformational states of ERβ induced by E2 and HT.
KW - Circular dichroism spectroscopy
KW - Conformational transitions
KW - Estrogen receptor beta
KW - Estrogen response element
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U2 - 10.1139/O06-144
DO - 10.1139/O06-144
M3 - Article
C2 - 17464340
AN - SCOPUS:34249940243
SN - 0829-8211
VL - 85
SP - 1
EP - 10
JO - Biochemistry and Cell Biology
JF - Biochemistry and Cell Biology
IS - 1
ER -