A novel point mutation in the DNA-binding domain (DBD) of the human glucocorticoid receptor causes primary generalized glucocorticoid resistance by disrupting the hydrophobic structure of its DBD

Michael L. Roberts, Tomoshige Kino, Nicolas C. Nicolaides, Darrell E. Hurt, Eleni Katsantoni, Amalia Sertedaki, Filadelfia Komianou, Korina Kassiou, George P. Chrousos, Evangelia Charmandari

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Context: Primary generalized glucocorticoid resistance is a rare genetic condition characterized by partial end-organ insensitivity to glucocorticoids. Most affected subjects present with clinical manifestations of mineralocorticoid and androgen excess. The condition has been associated with inactivating mutations in the human glucocorticoid receptor (hGR) gene, which impair the molecular mechanisms of hGRa action, thereby reducing tissue sensitivity to glucocorticoids. Objective: The aim of our study was to investigate the molecular mechanisms through which one previously described natural heterozygous V423A mutation, the second mutation detected in the DNA-binding domain (DBD) of the hGRa, affects glucocorticoid signal transduction. Design and Results: Compared with the wild-type receptor, hGRaV423A demonstrated a 72% reduction in its ability to trans activate the glucocorticoid-inducible mouse mammary tumor virus promoter in response to dexamethasone. The hGRaV423A receptor showed a significant reduction in its ability to bind to glucocorticoid-response elements of glucocorticoid-responsive genes, owing to structural alterations of the DBD confirmed by computer-based structural analysis. In addition, hGRaV423A demonstrated a 2.6-fold delay in nuclear translocation following exposure to the ligand, although it did not exert a dominant negative effect on the wild-type hGRa, had a similar affinity to the ligand with the wild-type receptor, and displayed a normal interaction with the GRIP1 coactivator in vitro. Conclusions: The natural mutant receptor hGRaV423A causes primary generalized glucocorticoid resistance by affecting multiple steps in the cascade of glucocorticoid receptor action, which primarily involve decreased ability to bind to target glucocorticoid response elements and delayed translocation into the nucleus.

Original languageEnglish
JournalJournal of Clinical Endocrinology and Metabolism
Volume98
Issue number4
DOIs
Publication statusPublished - Apr 2013
Externally publishedYes

Fingerprint

Glucocorticoid Receptors
Point Mutation
Glucocorticoids
DNA
Response Elements
Mutation
Ligands
Mouse mammary tumor virus
Mineralocorticoids
Genes
Glucocorticoid Receptor Deficiency
Signal transduction
Carcinogens
Dexamethasone
Androgens
Signal Transduction
Viruses
Structural analysis
Tumors
Tissue

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Endocrinology
  • Biochemistry, medical
  • Endocrinology, Diabetes and Metabolism

Cite this

A novel point mutation in the DNA-binding domain (DBD) of the human glucocorticoid receptor causes primary generalized glucocorticoid resistance by disrupting the hydrophobic structure of its DBD. / Roberts, Michael L.; Kino, Tomoshige; Nicolaides, Nicolas C.; Hurt, Darrell E.; Katsantoni, Eleni; Sertedaki, Amalia; Komianou, Filadelfia; Kassiou, Korina; Chrousos, George P.; Charmandari, Evangelia.

In: Journal of Clinical Endocrinology and Metabolism, Vol. 98, No. 4, 04.2013.

Research output: Contribution to journalArticle

Roberts, Michael L. ; Kino, Tomoshige ; Nicolaides, Nicolas C. ; Hurt, Darrell E. ; Katsantoni, Eleni ; Sertedaki, Amalia ; Komianou, Filadelfia ; Kassiou, Korina ; Chrousos, George P. ; Charmandari, Evangelia. / A novel point mutation in the DNA-binding domain (DBD) of the human glucocorticoid receptor causes primary generalized glucocorticoid resistance by disrupting the hydrophobic structure of its DBD. In: Journal of Clinical Endocrinology and Metabolism. 2013 ; Vol. 98, No. 4.
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abstract = "Context: Primary generalized glucocorticoid resistance is a rare genetic condition characterized by partial end-organ insensitivity to glucocorticoids. Most affected subjects present with clinical manifestations of mineralocorticoid and androgen excess. The condition has been associated with inactivating mutations in the human glucocorticoid receptor (hGR) gene, which impair the molecular mechanisms of hGRa action, thereby reducing tissue sensitivity to glucocorticoids. Objective: The aim of our study was to investigate the molecular mechanisms through which one previously described natural heterozygous V423A mutation, the second mutation detected in the DNA-binding domain (DBD) of the hGRa, affects glucocorticoid signal transduction. Design and Results: Compared with the wild-type receptor, hGRaV423A demonstrated a 72{\%} reduction in its ability to trans activate the glucocorticoid-inducible mouse mammary tumor virus promoter in response to dexamethasone. The hGRaV423A receptor showed a significant reduction in its ability to bind to glucocorticoid-response elements of glucocorticoid-responsive genes, owing to structural alterations of the DBD confirmed by computer-based structural analysis. In addition, hGRaV423A demonstrated a 2.6-fold delay in nuclear translocation following exposure to the ligand, although it did not exert a dominant negative effect on the wild-type hGRa, had a similar affinity to the ligand with the wild-type receptor, and displayed a normal interaction with the GRIP1 coactivator in vitro. Conclusions: The natural mutant receptor hGRaV423A causes primary generalized glucocorticoid resistance by affecting multiple steps in the cascade of glucocorticoid receptor action, which primarily involve decreased ability to bind to target glucocorticoid response elements and delayed translocation into the nucleus.",
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T1 - A novel point mutation in the DNA-binding domain (DBD) of the human glucocorticoid receptor causes primary generalized glucocorticoid resistance by disrupting the hydrophobic structure of its DBD

AU - Roberts, Michael L.

AU - Kino, Tomoshige

AU - Nicolaides, Nicolas C.

AU - Hurt, Darrell E.

AU - Katsantoni, Eleni

AU - Sertedaki, Amalia

AU - Komianou, Filadelfia

AU - Kassiou, Korina

AU - Chrousos, George P.

AU - Charmandari, Evangelia

PY - 2013/4

Y1 - 2013/4

N2 - Context: Primary generalized glucocorticoid resistance is a rare genetic condition characterized by partial end-organ insensitivity to glucocorticoids. Most affected subjects present with clinical manifestations of mineralocorticoid and androgen excess. The condition has been associated with inactivating mutations in the human glucocorticoid receptor (hGR) gene, which impair the molecular mechanisms of hGRa action, thereby reducing tissue sensitivity to glucocorticoids. Objective: The aim of our study was to investigate the molecular mechanisms through which one previously described natural heterozygous V423A mutation, the second mutation detected in the DNA-binding domain (DBD) of the hGRa, affects glucocorticoid signal transduction. Design and Results: Compared with the wild-type receptor, hGRaV423A demonstrated a 72% reduction in its ability to trans activate the glucocorticoid-inducible mouse mammary tumor virus promoter in response to dexamethasone. The hGRaV423A receptor showed a significant reduction in its ability to bind to glucocorticoid-response elements of glucocorticoid-responsive genes, owing to structural alterations of the DBD confirmed by computer-based structural analysis. In addition, hGRaV423A demonstrated a 2.6-fold delay in nuclear translocation following exposure to the ligand, although it did not exert a dominant negative effect on the wild-type hGRa, had a similar affinity to the ligand with the wild-type receptor, and displayed a normal interaction with the GRIP1 coactivator in vitro. Conclusions: The natural mutant receptor hGRaV423A causes primary generalized glucocorticoid resistance by affecting multiple steps in the cascade of glucocorticoid receptor action, which primarily involve decreased ability to bind to target glucocorticoid response elements and delayed translocation into the nucleus.

AB - Context: Primary generalized glucocorticoid resistance is a rare genetic condition characterized by partial end-organ insensitivity to glucocorticoids. Most affected subjects present with clinical manifestations of mineralocorticoid and androgen excess. The condition has been associated with inactivating mutations in the human glucocorticoid receptor (hGR) gene, which impair the molecular mechanisms of hGRa action, thereby reducing tissue sensitivity to glucocorticoids. Objective: The aim of our study was to investigate the molecular mechanisms through which one previously described natural heterozygous V423A mutation, the second mutation detected in the DNA-binding domain (DBD) of the hGRa, affects glucocorticoid signal transduction. Design and Results: Compared with the wild-type receptor, hGRaV423A demonstrated a 72% reduction in its ability to trans activate the glucocorticoid-inducible mouse mammary tumor virus promoter in response to dexamethasone. The hGRaV423A receptor showed a significant reduction in its ability to bind to glucocorticoid-response elements of glucocorticoid-responsive genes, owing to structural alterations of the DBD confirmed by computer-based structural analysis. In addition, hGRaV423A demonstrated a 2.6-fold delay in nuclear translocation following exposure to the ligand, although it did not exert a dominant negative effect on the wild-type hGRa, had a similar affinity to the ligand with the wild-type receptor, and displayed a normal interaction with the GRIP1 coactivator in vitro. Conclusions: The natural mutant receptor hGRaV423A causes primary generalized glucocorticoid resistance by affecting multiple steps in the cascade of glucocorticoid receptor action, which primarily involve decreased ability to bind to target glucocorticoid response elements and delayed translocation into the nucleus.

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