Point mutation I634A in the glucocorticoid receptor causes embryonic lethality by reduced ligand binding
Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved..
The glucocorticoid (GC) receptor (GR) is essential for normal development and in the initiation of inflammation. Healthy GRdim/dim mice with reduced dimerization propensity due to a point mutation (A465T) at the dimer interface of the GR DNA-binding domain (DBD) (here GRD/D) have previously helped to define the functions of GR monomers and dimers. Since GRD/D retains residual dimerization capacity, here we generated the dimer-nullifying double mutant GRD+L/D+L mice, featuring an additional mutation (I634A) in the ligand-binding domain (LBD) of GR. These mice are perinatally lethal, as are GRL/L mice (these mice have the I634A mutation but not the A465T mutation), displaying improper lung and skin formation. Using embryonic fibroblasts, high and low doses of dexamethasone (Dex), nuclear translocation assays, RNAseq, dimerization assays, and ligand-binding assays (and Kd values), we found that the lethal phenotype in these mice is due to insufficient ligand binding. These data suggest there is some correlation between GR dimerization potential and ligand affinity. We conclude that even a mutation as subtle as I634A, at a position not directly involved in ligand interactions sensu stricto, can still influence ligand binding and have a lethal outcome.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:298 |
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| Enthalten in: |
The Journal of biological chemistry - 298(2022), 2 vom: 15. Feb., Seite 101574 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Timmermans, Steven [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 28.04.2022 Date Revised 31.05.2022 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.jbc.2022.101574 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
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| 100 | 1 | |a Timmermans, Steven |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Point mutation I634A in the glucocorticoid receptor causes embryonic lethality by reduced ligand binding |
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| 500 | |a Date Completed 28.04.2022 | ||
| 500 | |a Date Revised 31.05.2022 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved. | ||
| 520 | |a The glucocorticoid (GC) receptor (GR) is essential for normal development and in the initiation of inflammation. Healthy GRdim/dim mice with reduced dimerization propensity due to a point mutation (A465T) at the dimer interface of the GR DNA-binding domain (DBD) (here GRD/D) have previously helped to define the functions of GR monomers and dimers. Since GRD/D retains residual dimerization capacity, here we generated the dimer-nullifying double mutant GRD+L/D+L mice, featuring an additional mutation (I634A) in the ligand-binding domain (LBD) of GR. These mice are perinatally lethal, as are GRL/L mice (these mice have the I634A mutation but not the A465T mutation), displaying improper lung and skin formation. Using embryonic fibroblasts, high and low doses of dexamethasone (Dex), nuclear translocation assays, RNAseq, dimerization assays, and ligand-binding assays (and Kd values), we found that the lethal phenotype in these mice is due to insufficient ligand binding. These data suggest there is some correlation between GR dimerization potential and ligand affinity. We conclude that even a mutation as subtle as I634A, at a position not directly involved in ligand interactions sensu stricto, can still influence ligand binding and have a lethal outcome | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a A465T | |
| 650 | 4 | |a I634A | |
| 650 | 4 | |a dimerization | |
| 650 | 4 | |a glucocorticoid receptor | |
| 650 | 4 | |a ligand binding | |
| 650 | 4 | |a mutational study | |
| 650 | 7 | |a Glucocorticoids |2 NLM | |
| 650 | 7 | |a Ligands |2 NLM | |
| 650 | 7 | |a Receptors, Glucocorticoid |2 NLM | |
| 650 | 7 | |a Dexamethasone |2 NLM | |
| 650 | 7 | |a 7S5I7G3JQL |2 NLM | |
| 700 | 1 | |a Verhoog, Nicolette J D |e verfasserin |4 aut | |
| 700 | 1 | |a Van Looveren, Kelly |e verfasserin |4 aut | |
| 700 | 1 | |a Dewaele, Sylviane |e verfasserin |4 aut | |
| 700 | 1 | |a Hochepied, Tino |e verfasserin |4 aut | |
| 700 | 1 | |a Eggermont, Melanie |e verfasserin |4 aut | |
| 700 | 1 | |a Gilbert, Barbara |e verfasserin |4 aut | |
| 700 | 1 | |a Boerema-de Munck, Anne |e verfasserin |4 aut | |
| 700 | 1 | |a Vanderhaeghen, Tineke |e verfasserin |4 aut | |
| 700 | 1 | |a Vanden Berghe, Joke |e verfasserin |4 aut | |
| 700 | 1 | |a Garcia Gonzalez, Natalia |e verfasserin |4 aut | |
| 700 | 1 | |a Vandewalle, Jolien |e verfasserin |4 aut | |
| 700 | 1 | |a Bloch, Yehudi |e verfasserin |4 aut | |
| 700 | 1 | |a Provost, Mathias |e verfasserin |4 aut | |
| 700 | 1 | |a Savvides, Savvas N |e verfasserin |4 aut | |
| 700 | 1 | |a De Bosscher, Karolien |e verfasserin |4 aut | |
| 700 | 1 | |a Declercq, Wim |e verfasserin |4 aut | |
| 700 | 1 | |a Rottier, Robbert J |e verfasserin |4 aut | |
| 700 | 1 | |a Louw, Ann |e verfasserin |4 aut | |
| 700 | 1 | |a Libert, Claude |e verfasserin |4 aut | |
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