Reciprocal mutagenesis between human α(L349, M528) and rainbow trout (M317, I496) estrogen receptor residues demonstrates their importance in ligand binding and gene expression at different temperatures

被引:17
作者
Matthews, JB
Clemons, JH
Zacharewski, TR
机构
[1] Michigan State Univ, Inst Environm Toxicol, Dept Biochem & Mol Biol, E Lansing, MI 48824 USA
[2] Michigan State Univ, Natl Food Safety & Toxicol Ctr, E Lansing, MI 48824 USA
关键词
estrogen receptor mutation; temperature instability; rainbow trout; transactivation; nuclear receptor;
D O I
10.1016/S0303-7207(01)00586-X
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Several fish proteins exhibit compromised function at temperatures outside of their normal physiological range, In this study. the effect of temperature on the ligand binding and the transactivation abilities of the rainbow trout estrogen receptor (rtER) and human estrogen receptor alpha (hER alpha) were examined. Saturation analysis and gene expression assays, using GST-ER and Gal4-ER fusion proteins consisting of the D, E and F domains of human (hER alpha def) and rainbow trout (rtERdef) receptors, show that GST-rtERdef E2 binding affinity and transactivation ability decrease with increasing temperature. A comparison of the amino acid sequence differences between their ligand binding pockets identified two conservative amino acid residue substitutions in rtER (M317, 1496) and hER alpha (L349, M528). The effect of these substitutions on ligand binding and transactivation were examined by constructing reciprocal mutants, which effectively exchanged the binding pockets between rtER and hER alpha. The rtERdef M317L:1496M double mutant exhibited increased E2 binding affinity and transactivation ability at higher temperatures, and displayed hER alpha phenotypic behavior for the phytoestrogen. coumestrol. The hER alpha def L349M:M528I double mutant also exhibited a modest trend towards adopting the rtER phenotype. These studies demonstrate that conservative changes in residue hydrophobicity and volume can significantly affect ER ligand binding and transactivation ability in a temperature-dependent manner. The lack of a complete exchange of phenotypes between rtER and hER alpha indicates that factors outside of the ligand binding pocket are also involved. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.
引用
收藏
页码:127 / 139
页数:13
相关论文
共 44 条
  • [1] Assessing the estrogenic and dioxin-like activities of chemicals and complex mixtures using in vitro recombinant receptor-reporter gene assays
    Balaguer, P
    Joyeux, A
    Denison, MS
    Vincent, R
    Gillesby, BE
    Zacharewski, T
    [J]. CANADIAN JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY, 1996, 74 (02) : 216 - 222
  • [2] TRANSCRIPTIONAL CONTROL BY NUCLEAR RECEPTORS
    BEATO, M
    [J]. FASEB JOURNAL, 1991, 5 (07) : 2044 - 2051
  • [3] A SINGLE AMINO-ACID THAT DETERMINES THE SENSITIVITY OF PROGESTERONE RECEPTORS TO RU486
    BENHAMOU, B
    GARCIA, T
    LEROUGE, T
    VERGEZAC, A
    GOFFLO, D
    BIGOGNE, C
    CHAMBON, P
    GRONEMEYER, H
    [J]. SCIENCE, 1992, 255 (5041) : 206 - 209
  • [4] BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
  • [5] BRASIER AR, 1989, BIOTECHNIQUES, V7, P1116
  • [6] Molecular basis of agonism and antagonism in the oestrogen receptor
    Brzozowski, AM
    Pike, ACW
    Dauter, Z
    Hubbard, RE
    Bonn, T
    Engstrom, O
    Ohman, L
    Greene, GL
    Gustafsson, JA
    Carlquist, M
    [J]. NATURE, 1997, 389 (6652) : 753 - 758
  • [7] STEROID-RECEPTOR FAMILY - STRUCTURE AND FUNCTIONS
    CARSONJURICA, MA
    SCHRADER, WT
    OMALLEY, BW
    [J]. ENDOCRINE REVIEWS, 1990, 11 (02) : 201 - 220
  • [8] DIFFERENT THERMOSTABILITIES OF SARCOPLASMIC-RETICULUM (CA2++MG2+)-ATPASES FROM RABBIT AND TROUT MUSCLES
    DETOLEDO, FGS
    ALBUQUERQUE, MC
    GOULART, BH
    CHINI, EN
    [J]. COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-PHARMACOLOGY TOXICOLOGY & ENDOCRINOLOGY, 1995, 111 (01): : 93 - 98
  • [9] Estrogenic activity of chlordecone, o,p'-DDT and o,p'-DDE in juvenile rainbow trout: Induction of vitellogenesis and interaction with hepatic estrogen binding sites
    Donohoe, RM
    Curtis, LR
    [J]. AQUATIC TOXICOLOGY, 1996, 36 (1-2) : 31 - 52
  • [10] Different residues of the human estrogen receptor are involved in the recognition of structurally diverse estrogens and antiestrogens
    Ekena, K
    Weis, KE
    Katzenellenbogen, JA
    Katzenellenbogen, BS
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (08) : 5069 - 5075