Structural basis for nuclear hormone receptor DNA binding

Helsen, Christine; Kerkhofs, Stefanie; Clinckemalie, Liesbeth; Spans, Lien; Laurent, Michaël; Boonen, Steven; Vanderschueren, Dirk; Claessens, Frank

doi:10.1016/j.mce.2011.07.025

Cited by 127 publications

(89 citation statements)

References 77 publications

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“…17β-Estradiol (E2) association with the ERα ligand binding domain (LBD) drives large conformational changes (10) resulting in ERα dissociation from chaperones (11, 12), unmasking of domains for receptor dimerization, and DNA binding (13,14). Whereas the LBD contains the main dimerization domain (15), the extreme C-terminal domain of the receptor (F domain) imposes a restraint on dimerization (15, 16), although the regulation of this remains fully elusive.…”

mentioning

confidence: 99%

Interaction of 14-3-3 proteins with the Estrogen Receptor Alpha F domain provides a drug target interface

Leeuwen

Pereira

Flach

et al. 2013

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

126

180

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Estrogen receptor alpha (ERα) is involved in numerous physiological and pathological processes, including breast cancer. Breast cancer therapy is therefore currently directed at inhibiting the transcriptional potency of ERα, either by blocking estrogen production through aromatase inhibitors or antiestrogens that compete for hormone binding. Due to resistance, new treatment modalities are needed and as ERα dimerization is essential for its activity, interference with receptor dimerization offers a new opportunity to exploit in drug design. Here we describe a unique mechanism of how ERα dimerization is negatively controlled by interaction with 14-3-3 proteins at the extreme C terminus of the receptor. Moreover, the small-molecule fusicoccin (FC) stabilizes this ERα/14-3-3 interaction. Cocrystallization of the trimeric ERα/ 14-3-3/FC complex provides the structural basis for this stabilization and shows the importance of phosphorylation of the penultimate Threonine (ERα-T 594 ) for high-affinity interaction. We confirm that T 594 is a distinct ERα phosphorylation site in the breast cancer cell line MCF-7 using a phospho-T 594 -specific antibody and by mass spectrometry. In line with its ERα/14-3-3 interaction stabilizing effect, fusicoccin reduces the estradiol-stimulated ERα dimerization, inhibits ERα/chromatin interactions and downstream gene expression, resulting in decreased cell proliferation. Herewith, a unique functional phosphosite and an alternative regulation mechanism of ERα are provided, together with a small molecule that selectively targets this ERα/14-3-3 interface.T he estrogen receptor alpha (ERα) is a ligand-dependent transcription factor and the driving force of cell proliferation in 75% of all breast cancers. Current therapeutic strategies to treat these tumors rely on selective ER modulators (SERMs), like tamoxifen (TAM) (1) or aromatase inhibitors (AIs) that block estradiol synthesis (2). Although the benefits of treating hormone-sensitive breast cancers with SERMs and AIs are evident, resistance to treatment is commonly observed (3, 4). To overcome resistance, selective ERα down-regulators (SERDs) can for instance be applied that inhibit ERα signaling through receptor degradation (5, 6). Approaches that target the ERα/ DNA or ERα/cofactor interactions are explored as well (5, 7), but other essential steps in the ERα activation cascade are currently unexploited in drug design, also due to a lack of molecular understanding of the processes at hand.One such step that is crucial for many aspects of ERα functioning is ligand-driven receptor dimerization (8, 9). 17β-Estradiol (E2) association with the ERα ligand binding domain (LBD) drives large conformational changes (10) resulting in ERα dissociation from chaperones (11, 12), unmasking of domains for receptor dimerization, and DNA binding (13,14). Whereas the LBD contains the main dimerization domain (15), the extreme C-terminal domain of the receptor (F domain) imposes a restraint on dimerization (15, 16), although the regulation of this remain...

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mentioning

confidence: 99%

Interaction of 14-3-3 proteins with the Estrogen Receptor Alpha F domain provides a drug target interface

Leeuwen

Pereira

Flach

et al. 2013

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

126

180

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“…The DBD is the most conserved NR domain and consists of two ␣ helices that are coordinated by zinc mole-cules, thus forming two zinc-finger modules. The first zinc finger is responsible for the recognition of AREs, while the second zinc finger is involved in DNA-dependent dimerization (31,40,51).…”

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confidence: 99%

Evidence for DNA-Binding Domain–Ligand-Binding Domain Communications in the Androgen Receptor

Helsen

Dubois

Verfaillie

et al. 2012

Molecular and Cellular Biology

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DNA binding as well as ligand binding by nuclear receptors has been studied extensively. Both binding functions are attributed to isolated domains of which the structure is known. The crystal structure of a complete receptor in complex with its ligand and DNA-response element, however, has been solved only for the peroxisome proliferator-activated receptor ␥ (PPAR␥)-retinoid X receptor ␣ (RXR␣) heterodimer. This structure provided the first indication of direct interactions between the DNA-binding domain (DBD) and ligand-binding domain (LBD). In this study, we investigated whether there is a similar interface between the DNA-and ligand-binding domains for the androgen receptor (AR). Despite the structural differences between the AR-and PPAR␥-LBD, a combination of in silico modeling and docking pointed out a putative interface between AR-DBD and AR-LBD. The surfaces were subjected to a point mutation analysis, which was inspired by known AR mutations described in androgen insensitivity syndromes and prostate cancer. Surprisingly, AR-LBD mutations D695N, R710A, F754S, and P766A induced a decrease in DNA binding but left ligand binding unaffected, while the DBD-residing mutations K590A, K592A, and E621A lowered the ligand-binding but not the DNA-binding affinity. We therefore propose that these residues are involved in allosteric communications between the AR-DBD and AR-LBD. N uclear receptors (NRs) are involved in many physiological processes, diseases, and therapeutic applications. They are transcription factors that contain a DNA-binding domain (DBD) composed of 2 zinc fingers (40) and a ligand-binding domain (LBD) formed by 12 ␣ helices (60). The structures of the separate DNA-binding and ligand-binding domains of many receptors have already revealed a large amount of information. The structure of the LBD has especially led to a more focused search for new agonists and antagonists for many therapeutic indications where NRs are involved. The exact structure of full-size NRs bound to DNA and ligand will help us in understanding the basic mechanism of nuclear receptor signaling but will also provide new targets for therapeutic strategies. The coordinates of the peroxisome proliferator-activated receptor ␥ (PPAR␥)-retinoid X receptor ␣ (RXR␣) cocrystal bound as a heterodimer to the DNA have been reported (11) and revealed a contact between PPAR␥ and RXR␣ more intimate than expected. The existence of the previously unknown interface between PPAR␥-LBD and RXR␣-DBD was corroborated with a mutation analysis. Since this has not yet been confirmed by other techniques like small-angle X-ray scattering (SAXS) and small-angle neutron scattering (SANS) (49), it is debated whether such a communication exists. There is, however, strong evidence for allosteric communications, e.g., between the DBD and LBD, in nuclear receptors. A most remarkable observation was made for the glucocorticoid receptor (GR) when slightly different response elements were tested in gene reporter assays: small changes in the DNA sequence had an importa...

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“…Also, GR may substitute for AR signalling. The two receptors can bind to the same sequence-specific response elements (Helsen et al 2012) and many AR-regulated genes are also regulated by GR. Indeed, GR is reported to induce KLK3 and TMPRSS2 expression (Tomlins et al 2006, Tran et al 2009, Arora et al 2013, Isikbay et al 2014.…”

Section: Glucocorticoid Receptor (Gr or Nr3c1)mentioning

confidence: 99%

WOMEN IN CANCER THEMATIC REVIEW: New roles for nuclear receptors in prostate cancer

Leach

Powell

Bevan

2016

Endocrine-Related Cancer

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Prostate cancer has, for decades, been treated by inhibiting androgen signalling. This is effective in the majority of patients, but inevitably resistance develops and patients progress to life-threatening metastatic disease -hence the quest for new effective therapies for 'castrate-resistant' prostate cancer (CRPC). Studies into what pathways can drive tumour recurrence under these conditions has identified several other nuclear receptor signalling pathways as potential drivers or modulators of CRPC. The nuclear receptors constitute a large (48 members) superfamily of transcription factors sharing a common modular functional structure. Many of them are activated by the binding of small lipophilic molecules, making them potentially druggable. Even those for which no ligand exists or has yet been identified may be tractable to activity modulation by small molecules. Moreover, genomic studies have shown that in models of CRPC, other nuclear receptors can potentially drive similar transcriptional responses to the androgen receptor, while analysis of expression and sequencing databases shows disproportionately high mutation and copy number variation rates among the superfamily. Hence, the nuclear receptor superfamily is of intense interest in the drive to understand how prostate cancer recurs and how we may best treat such recurrent disease. This review aims to provide a snapshot of the current knowledge of the roles of different nuclear receptors in prostate cancer -a rapidly evolving field of research.

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Structural basis for nuclear hormone receptor DNA binding

Cited by 127 publications

References 77 publications

Interaction of 14-3-3 proteins with the Estrogen Receptor Alpha F domain provides a drug target interface

Interaction of 14-3-3 proteins with the Estrogen Receptor Alpha F domain provides a drug target interface

Evidence for DNA-Binding Domain–Ligand-Binding Domain Communications in the Androgen Receptor

WOMEN IN CANCER THEMATIC REVIEW: New roles for nuclear receptors in prostate cancer

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