An antiestrogen-responsive estrogen receptor-α mutant (D351Y) shows weak AF-2 activity in the presence of tamoxifen

Paul Webb, Phuong Nguyen, Cathleen Valentine, Ross V. Weatherman, Thomas (Tom) Scanlan, Peter J. Kushner

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Antiestrogens, including tamoxifen and raloxifene, block estrogen receptor (ER) action by blocking the interactions of an estrogen-dependent activation function (AF-2) with p160 coactivators. Although tamoxifen does show some agonist activity in the presence of ERα, this stems from a distinct constitutive activation function (AF-1) that lies within the ERα N terminus. Previous studies identified a naturally occurring mutation (D351Y) that allows ERα to perceive tamoxifen and raloxifene as estrogens. Here, we examine the contributions of ERα activation functions to the D351Y pheno-type. We find that the AF-2 function of ERα D351Y lacks detectable tamoxifen-dependent activity when tested in isolation but does synergize with AF-1 to allow enhanced tamoxifen response. Weak tamoxifen-dependent interactions between the ERα D351Y AF-2 function and GRIP1, a representative p160, can be detected in glutathione S-transferase binding assays and mammalian two-hybrid assays. Furthermore, tamoxifen-dependent AF-2 activity can be detected in the presence of ERα D351Y and high levels of overexpressed GRIP1. We therefore propose that the D351Y mutation allows weak tamoxifen-dependent AF-2 activity but that this activity is only detectable when AF-1 is strong, and AF-1 and AF-2 synergize, or when p160s are overexpressed. We discuss the possible structural basis of this effect.

Original languageEnglish (US)
Pages (from-to)37552-37558
Number of pages7
JournalJournal of Biological Chemistry
Volume275
Issue number48
DOIs
StatePublished - Dec 1 2000
Externally publishedYes

Fingerprint

Furylfuramide
Estrogen Receptor Modulators
Tamoxifen
Estrogen Receptors
Chemical activation
Assays
Estrogens
Two-Hybrid System Techniques
Mutation
Glutathione Transferase

ASJC Scopus subject areas

  • Biochemistry

Cite this

An antiestrogen-responsive estrogen receptor-α mutant (D351Y) shows weak AF-2 activity in the presence of tamoxifen. / Webb, Paul; Nguyen, Phuong; Valentine, Cathleen; Weatherman, Ross V.; Scanlan, Thomas (Tom); Kushner, Peter J.

In: Journal of Biological Chemistry, Vol. 275, No. 48, 01.12.2000, p. 37552-37558.

Research output: Contribution to journalArticle

Webb, Paul ; Nguyen, Phuong ; Valentine, Cathleen ; Weatherman, Ross V. ; Scanlan, Thomas (Tom) ; Kushner, Peter J. / An antiestrogen-responsive estrogen receptor-α mutant (D351Y) shows weak AF-2 activity in the presence of tamoxifen. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 48. pp. 37552-37558.
@article{0faa9f3c1dfe4fc6a462be28b28f4077,
title = "An antiestrogen-responsive estrogen receptor-α mutant (D351Y) shows weak AF-2 activity in the presence of tamoxifen",
abstract = "Antiestrogens, including tamoxifen and raloxifene, block estrogen receptor (ER) action by blocking the interactions of an estrogen-dependent activation function (AF-2) with p160 coactivators. Although tamoxifen does show some agonist activity in the presence of ERα, this stems from a distinct constitutive activation function (AF-1) that lies within the ERα N terminus. Previous studies identified a naturally occurring mutation (D351Y) that allows ERα to perceive tamoxifen and raloxifene as estrogens. Here, we examine the contributions of ERα activation functions to the D351Y pheno-type. We find that the AF-2 function of ERα D351Y lacks detectable tamoxifen-dependent activity when tested in isolation but does synergize with AF-1 to allow enhanced tamoxifen response. Weak tamoxifen-dependent interactions between the ERα D351Y AF-2 function and GRIP1, a representative p160, can be detected in glutathione S-transferase binding assays and mammalian two-hybrid assays. Furthermore, tamoxifen-dependent AF-2 activity can be detected in the presence of ERα D351Y and high levels of overexpressed GRIP1. We therefore propose that the D351Y mutation allows weak tamoxifen-dependent AF-2 activity but that this activity is only detectable when AF-1 is strong, and AF-1 and AF-2 synergize, or when p160s are overexpressed. We discuss the possible structural basis of this effect.",
author = "Paul Webb and Phuong Nguyen and Cathleen Valentine and Weatherman, {Ross V.} and Scanlan, {Thomas (Tom)} and Kushner, {Peter J.}",
year = "2000",
month = "12",
day = "1",
doi = "10.1074/jbc.M007435200",
language = "English (US)",
volume = "275",
pages = "37552--37558",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "48",

}

TY - JOUR

T1 - An antiestrogen-responsive estrogen receptor-α mutant (D351Y) shows weak AF-2 activity in the presence of tamoxifen

AU - Webb, Paul

AU - Nguyen, Phuong

AU - Valentine, Cathleen

AU - Weatherman, Ross V.

AU - Scanlan, Thomas (Tom)

AU - Kushner, Peter J.

PY - 2000/12/1

Y1 - 2000/12/1

N2 - Antiestrogens, including tamoxifen and raloxifene, block estrogen receptor (ER) action by blocking the interactions of an estrogen-dependent activation function (AF-2) with p160 coactivators. Although tamoxifen does show some agonist activity in the presence of ERα, this stems from a distinct constitutive activation function (AF-1) that lies within the ERα N terminus. Previous studies identified a naturally occurring mutation (D351Y) that allows ERα to perceive tamoxifen and raloxifene as estrogens. Here, we examine the contributions of ERα activation functions to the D351Y pheno-type. We find that the AF-2 function of ERα D351Y lacks detectable tamoxifen-dependent activity when tested in isolation but does synergize with AF-1 to allow enhanced tamoxifen response. Weak tamoxifen-dependent interactions between the ERα D351Y AF-2 function and GRIP1, a representative p160, can be detected in glutathione S-transferase binding assays and mammalian two-hybrid assays. Furthermore, tamoxifen-dependent AF-2 activity can be detected in the presence of ERα D351Y and high levels of overexpressed GRIP1. We therefore propose that the D351Y mutation allows weak tamoxifen-dependent AF-2 activity but that this activity is only detectable when AF-1 is strong, and AF-1 and AF-2 synergize, or when p160s are overexpressed. We discuss the possible structural basis of this effect.

AB - Antiestrogens, including tamoxifen and raloxifene, block estrogen receptor (ER) action by blocking the interactions of an estrogen-dependent activation function (AF-2) with p160 coactivators. Although tamoxifen does show some agonist activity in the presence of ERα, this stems from a distinct constitutive activation function (AF-1) that lies within the ERα N terminus. Previous studies identified a naturally occurring mutation (D351Y) that allows ERα to perceive tamoxifen and raloxifene as estrogens. Here, we examine the contributions of ERα activation functions to the D351Y pheno-type. We find that the AF-2 function of ERα D351Y lacks detectable tamoxifen-dependent activity when tested in isolation but does synergize with AF-1 to allow enhanced tamoxifen response. Weak tamoxifen-dependent interactions between the ERα D351Y AF-2 function and GRIP1, a representative p160, can be detected in glutathione S-transferase binding assays and mammalian two-hybrid assays. Furthermore, tamoxifen-dependent AF-2 activity can be detected in the presence of ERα D351Y and high levels of overexpressed GRIP1. We therefore propose that the D351Y mutation allows weak tamoxifen-dependent AF-2 activity but that this activity is only detectable when AF-1 is strong, and AF-1 and AF-2 synergize, or when p160s are overexpressed. We discuss the possible structural basis of this effect.

UR - http://www.scopus.com/inward/record.url?scp=0034532014&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034532014&partnerID=8YFLogxK

U2 - 10.1074/jbc.M007435200

DO - 10.1074/jbc.M007435200

M3 - Article

C2 - 10986290

AN - SCOPUS:0034532014

VL - 275

SP - 37552

EP - 37558

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 48

ER -