Point mutation I634A in the glucocorticoid receptor causes embryonic lethality by reduced ligand binding

Steven Timmermans, Nicolette J.D. Verhoog, Kelly Van Looveren, Sylviane Dewaele, Tino Hochepied, Melanie Eggermont, Barbara Gilbert, Anne Boerema de Munck, Tineke Vanderhaeghen, Joke Vanden Berghe, Natalia Garcia Gonzalez, Jolien Vandewalle, Yehudi Bloch, Mathias Provost, Savvas N. Savvides, Karolien De Bosscher, Wim Declercq, Robbert J. Rottier, Ann Louw, Claude Libert*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

1 Citation (Scopus)
3 Downloads (Pure)

Abstract

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.

Original languageEnglish
Article number101574
JournalJournal of Biological Chemistry
Volume298
Issue number2
DOIs
Publication statusPublished - 1 Feb 2022

Bibliographical note

Funding Information:
We wish to thank Prof. Jan Tuckermann for providing GRD/D and GR-/- mice. We are thankful to animal house caretakers for animal care. We acknowledge the VIB Nucleomics Core for the RNA sequencing. We would like to thank the VIB Bio-Imaging Core for training, support, and access to the instrument park. Research in the author's laboratories was funded by the Agency for Innovation of Science and Technology in Flanders (IWT), the Research Council of Ghent University (GOA Program), the Research Foundation Flanders (FWO-Vlaanderen), and Flanders Institute for Biotechnology (VIB).

Funding Information:
Acknowledgments—We wish to thank Prof. Jan Tuckermann for providing GRD/D and GR−/− mice. We are thankful to animal house caretakers for animal care. We acknowledge the VIB Nucleomics Core for the RNA sequencing. We would like to thank the VIB Bio-Imaging Core for training, support, and access to the instrument park. Research in the author’s laboratories was funded by the Agency for Innovation of Science and Technology in Flanders (IWT), the Research Council of Ghent University (GOA Program), the Research Foundation Flanders (FWO-Vlaanderen), and Flanders Institute for Biotechnology (VIB).

Publisher Copyright:
© 2022 THE AUTHORS. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Fingerprint

Dive into the research topics of 'Point mutation I634A in the glucocorticoid receptor causes embryonic lethality by reduced ligand binding'. Together they form a unique fingerprint.

Cite this