A Conformational Switch in the Ligand-binding Domain Regulates the Dependence of the Glucocorticoid Receptor on Hsp90

Ricketson, Derek; Hostick, U.; Fang, L.; Yamamoto, Keith R.; Darimont, Beatrice

doi:10.1016/j.jmb.2007.02.057

Cited by 58 publications

(51 citation statements)

References 43 publications

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“…GR has a modular structure consisting of three major functional domains, the N-terminal domain (NTD), the central DNA-binding domain (DBD), and the C-terminal ligand-binding domain (LBD). Upon ligand binding, GR dissociates from multichaperone complexes, dimerizes, and translocates into the nucleus (5). In addition to its role in ligand recognition, the LBD contains a ligand-dependent activation function (AF-2) that is tightly regulated by hormone binding and several coactivators interacting with this domain (6).…”

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confidence: 99%

RSUME Enhances Glucocorticoid Receptor SUMOylation and Transcriptional Activity

Druker

Liberman

Antunica-Noguerol

et al. 2013

Molecular and Cellular Biology

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cGlucocorticoid receptor (GR) activity is modulated by posttranslational modifications, including phosphorylation, ubiquitination, and SUMOylation. The GR has three SUMOylation sites: lysine 297 (K297) and K313 in the N-terminal domain (NTD) and K721 within the ligand-binding domain. SUMOylation of the NTD sites mediates the negative effect of the synergy control motifs of GR on promoters with closely spaced GR binding sites. There is scarce evidence on the role of SUMO conjugation to K721 and its impact on GR transcriptional activity. We have previously shown that RSUME (RWD-containing SUMOylation enhancer) increases protein SUMOylation. We now demonstrate that RSUME interacts with the GR and increases its SUMOylation. RSUME regulates GR transcriptional activity and the expression of its endogenous target genes, FKBP51 and S100P. RSUME uncovers a positive role for the third SUMOylation site, K721, on GR-mediated transcription, demonstrating that GR SUMOylation acts positively in the presence of a SUMOylation enhancer. Both mutation of K721 and small interfering RNA-mediated RSUME knockdown diminish GRIP1 coactivator activity. RSUME, whose expression is induced under stress conditions, is a key factor in heat shock-induced GR SUMOylation. These results show that inhibitory and stimulatory SUMO sites are present in the GR and at higher SUMOylation levels the stimulatory one becomes dominant. G lucocorticoid (GC) actions are mediated by the glucocorticoid receptor (GR), a ligand-activated transcription factor and member of the nuclear receptor (NR) superfamily. GR plays an important role in gene transcription (1), regulating processes such as inflammation, glucose and lipid metabolism, stress response, development, the cell cycle, and apoptosis (2-4). GR has a modular structure consisting of three major functional domains, the N-terminal domain (NTD), the central DNA-binding domain (DBD), and the C-terminal ligand-binding domain (LBD). Upon ligand binding, GR dissociates from multichaperone complexes, dimerizes, and translocates into the nucleus (5). In addition to its role in ligand recognition, the LBD contains a ligand-dependent activation function (AF-2) that is tightly regulated by hormone binding and several coactivators interacting with this domain (6). GR-mediated transcriptional activity is regulated not only by its binding to GC response elements (GREs) and the balance between coactivators and corepressors but also by posttranslational modifications, which include phosphorylation, ubiquitination, and SUMOylation (7-12).The SUMO conjugation pathway has similarities to the ubiquitination process but uses a different set of enzymes involved in processing, attachment, and removal of SUMO (13,14). First, SUMO is activated in an ATP-dependent manner by the heterodimeric E1-activating enzyme SAE1/2. Activated SUMO is then transferred to the E2-conjugating enzyme Ubc9 and is conjugated to specific lysine residues in substrate proteins. Efficient SUMO conjugation in vivo further requires the action of specific SU...

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confidence: 99%

RSUME Enhances Glucocorticoid Receptor SUMOylation and Transcriptional Activity

Druker

Liberman

Antunica-Noguerol

et al. 2013

Molecular and Cellular Biology

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“…This revealed a very similar conformation to that seen with Dex, but there was a single difference, namely the addition of a patch of positive charge on the external surface of the LBD. Ricketson and co-workers were able to demonstrate, through amino acid substitution, that this surface is required for HSP90 interaction (Ricketson et al, 2007). HSP90 recognises the GR LBD through two, defined hydrophobic sites and binds to a solvent accessible major groove maintaining GR stability and permitting highaffinity ligand binding (Fang et al, 2006), as depicted in Fig.…”

Section: Discussionmentioning

confidence: 99%

“…The chaperone heat shock protein 90 (HSP90) is known to play key roles in this aspect of GR biology, including maintaining GR structure, ligand-binding activity, and trafficking of GR between nucleus and cytoplasm (Segnitz and Gehring, 1997;Tago et al, 2004;Kakar et al, 2006;Grad and Picard, 2007;Echeverría et al, 2009). GR residues identified by Ricketson and co-workers (Ricketson et al, 2007) as important for HSP90 interaction were mapped onto the crystal structure of GR bound to Dex (Fig. 6A).…”

Section: Structural Modelling Suggests That Nsgs Modify the Hsp90 Intmentioning

confidence: 99%

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A ligand-specific kinetic switch regulates glucocorticoid receptor trafficking and function

Trebble

Woolven

Saunders

et al. 2013

Journal of Cell Science

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SummaryThe ubiquitously expressed glucocorticoid receptor (GR) is a major drug target for inflammatory disease, but issues of specificity and target tissue sensitivity remain. We now identify high potency, non-steroidal GR ligands, GSK47867A and GSK47869A, which induce a novel conformation of the GR ligand-binding domain (LBD) and augment the efficacy of cellular action. Despite their high potency, GSK47867A and GSK47869A both induce surprisingly slow GR nuclear translocation, followed by prolonged nuclear GR retention, and transcriptional activity following washout. We reveal that GSK47867A and GSK47869A specifically alter the GR LBD structure at the HSP90-binding site. The alteration in the HSP90-binding site was accompanied by resistance to HSP90 antagonism, with persisting transactivation seen after geldanamycin treatment. Taken together, our studies reveal a new mechanism governing GR intracellular trafficking regulated by ligand binding that relies on a specific surface charge patch within the LBD. This conformational change permits extended GR action, probably because of altered GR-HSP90 interaction. This chemical series may offer anti-inflammatory drugs with prolonged duration of action due to altered pharmacodynamics rather than altered pharmacokinetics.

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“…A conformational change in the GR LBD that accompanies GC binding causes GR activation and nuclear translocation (9,10). Once in the nucleus, GR binds to the so-called positive GC response element [i.e., (+)GRE DNA-binding sequence (DBS)].…”

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confidence: 99%

A trilogy of glucocorticoid receptor actions

Wahli

2016

Proc. Natl. Acad. Sci. U.S.A.

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A Conformational Switch in the Ligand-binding Domain Regulates the Dependence of the Glucocorticoid Receptor on Hsp90

Cited by 58 publications

References 43 publications

RSUME Enhances Glucocorticoid Receptor SUMOylation and Transcriptional Activity

RSUME Enhances Glucocorticoid Receptor SUMOylation and Transcriptional Activity

A ligand-specific kinetic switch regulates glucocorticoid receptor trafficking and function

A trilogy of glucocorticoid receptor actions

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