NRIP, a Novel Nuclear Receptor Interaction Protein, Enhances the Transcriptional Activity of Nuclear Receptors

Abstract SMRT (silencing mediator of retinoic acid and thyroid hormone receptor) and N-CoR (nuclear receptor corepressor) are two related transcriptional corepressors that contain separable domains capable of interacting with unliganded nuclear receptors and repressing basal transcription. To decipher the mechanisms of receptor interaction and transcriptional repression by SMRT/N-CoR, we have characterized protein-protein interacting surfaces between SMRT and nuclear receptors and defined transcriptional repression domains of both SMRT and N-CoR. Deletional analysis reveals two individual nuclear receptor domains necessary for stable association with SMRT and a C-terminal helix essential for corepressor dissociation. Coordinately, two SMRT domains are found to interact independently with the receptors. Functional analysis reveals that SMRT contains two distinct repression domains, and the corresponding regions in N-CoR also repress basal transcription. Both repression domains in SMRT and N-CoR interact weakly with mSin3A, which in turn associates with a histone deacetylase HDAC1 in a mammalian two-hybrid assay. Far-Western analysis demonstrates a direct protein-protein interaction between two N-CoR repression domains with mSin3A. Finally we demonstrate that overexpression of full-length SMRT further represses basal transcription from natural promoters. Together, these results support a role of SMRT/N-CoR in corepression through the utilization of multiple mechanisms for receptor interactions and transcriptional repression.

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Inactivation of the Nuclear Receptor Coactivator RAP250 in Mice Results in Placental Vascular Dysfunction

Molecular and Cellular Biology ◽

10.1128/mcb.23.4.1260-1268.2003 ◽

2003 ◽

Vol 23 (4) ◽

pp. 1260-1268 ◽

Cited By ~ 72

Author(s):

Per Antonson ◽

Gertrud U. Schuster ◽

Ling Wang ◽

Björn Rozell ◽

Elin Holter ◽

...

Keyword(s):

Nuclear Receptors ◽

Nuclear Receptor ◽

Blood Circulation ◽

Transcriptional Activity ◽

Vascular Dysfunction ◽

Diverse Group ◽

Placental Development ◽

Nuclear Receptor Coactivator ◽

The Past ◽

Placental Blood

ABSTRACT Coactivators constitute a diverse group of proteins that are essential for optimal transcriptional activity of nuclear receptors. In the past few years many coactivators have been identified but it is still unclear whether these proteins interact indiscriminately with all nuclear receptors and whether there is some redundancy in their functions. We have previously cloned and characterized RAP250 (ASC-2/PRIP/TRBP/NRC), an LXXLL-containing coactivator for nuclear receptors. In order to study its biological role, Rap250 null mice were generated by gene targeting. Here we show that genetic disruption of Rap250 results in embryonic lethality at embryonic day (E) 13.5. Histological examination of placentas revealed a dramatically reduced spongiotrophoblast layer, a collapse of blood vessels in the region bordering the spongiotrophoblast, and labyrinthine layers in placentas from Rap250−/− embryos. These findings suggest that the lethality of Rap250−/− embryos is the result of obstructed placental blood circulation. Moreover, the transcriptional activity of PPARγ is reduced in fibroblasts derived from Rap250−/− embryos, suggesting that RAP250 is an essential coactivator for this nuclear receptor in the placenta. Our results demonstrate that RAP250 is necessary for placental development and thus essential for embryonic development.

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Development of Peptide Antagonists for the Androgen Receptor Using Combinatorial Peptide Phage Display

Molecular Endocrinology ◽

10.1210/me.2005-0072 ◽

2005 ◽

Vol 19 (10) ◽

pp. 2478-2490 ◽

Cited By ~ 53

Author(s):

Ching-yi Chang ◽

Jennifer Abdo ◽

Tanya Hartney ◽

Donald P. McDonnell

Keyword(s):

Androgen Receptor ◽

Phage Display ◽

Nuclear Receptors ◽

Nuclear Receptor ◽

Transcriptional Activity ◽

Binding Pocket ◽

Orphan Nuclear Receptors ◽

Nuclear Receptor Coactivator ◽

Pharmacological Significance ◽

Peptide Phage Display

Abstract Under the auspices of the Nuclear Receptor Signaling Atlas (NURSA) , we have undertaken to evaluate the feasibility of targeting nuclear receptor-coactivator surfaces for new drug discovery. The underlying objective of this approach is to provide the research community with reagents that can be used to modulate the transcriptional activity of nuclear receptors. Using combinatorial peptide phage display, we have been able to develop peptide antagonists that target specific nuclear receptor (NR)-coactivator binding surfaces. It can be appreciated that reagents of this nature will be of use in the study of orphan nuclear receptors for whom classical ligands have not yet been identified. In addition, because the interaction of coactivators with the receptor is an obligate step for NR transcriptional activity, it is anticipated that peptides that block these interactions will enable the definition of the biological and pharmacological significance of individual NR-coactivator interactions. In this report, we describe the use of this approach to develop antagonists of the androgen receptor by targeting its coactivator-binding pocket and their use to study the coactivator-binding surface of this receptor. Based on our findings, we believe that molecules that function by disrupting the androgen receptor-cofactor interactions will have use in the treatment of prostate cancer.

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Structural basis for heme-dependent NCoR binding to the transcriptional repressor REV-ERBβ

Science Advances ◽

10.1126/sciadv.abc6479 ◽

2021 ◽

Vol 7 (5) ◽

pp. eabc6479

Author(s):

Sarah A. Mosure ◽

Timothy S. Strutzenberg ◽

Jinsai Shang ◽

Paola Munoz-Tello ◽

Laura A. Solt ◽

...

Keyword(s):

Nuclear Receptors ◽

Binding Mode ◽

Structural Basis ◽

Receptor Interaction ◽

Heme Binding ◽

Endogenous Ligand ◽

Interaction Domain ◽

Response Elements ◽

Biochemical Assays ◽

Chemical Cross Linking

Heme is the endogenous ligand for the constitutively repressive REV-ERB nuclear receptors, REV-ERBα (NR1D1) and REV-ERBβ (NR1D2), but how heme regulates REV-ERB activity remains unclear. Cellular studies indicate that heme is required for the REV-ERBs to bind the corepressor NCoR and repress transcription. However, fluorescence-based biochemical assays suggest that heme displaces NCoR; here, we show that this is due to a heme-dependent artifact. Using ITC and NMR spectroscopy, we show that heme binding remodels the thermodynamic interaction profile of NCoR receptor interaction domain (RID) binding to REV-ERBβ ligand-binding domain (LBD). We solved two crystal structures of REV-ERBβ LBD cobound to heme and NCoR peptides, revealing the heme-dependent NCoR binding mode. ITC and chemical cross-linking mass spectrometry reveals a 2:1 LBD:RID stoichiometry, consistent with cellular studies showing that NCoR-dependent repression of REV-ERB transcription occurs on dimeric DNA response elements. Our findings should facilitate renewed progress toward understanding heme-dependent REV-ERB activity.

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Heme-binding to the nuclear receptor retinoid X receptor α (RXRα) leads to the inhibition of the transcriptional activity

Gene ◽

10.1016/j.gene.2008.07.006 ◽

2008 ◽

Vol 423 (2) ◽

pp. 207-214 ◽

Cited By ~ 9

Author(s):

Saki Gotoh ◽

Yoshiko Ohgari ◽

Takayuki Nakamura ◽

Takashi Osumi ◽

Shigeru Taketani

Keyword(s):

Nuclear Receptor ◽

Transcriptional Activity ◽

Retinoid X Receptor ◽

Heme Binding

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Dopamine neurons express multiple isoforms of the nuclear receptor nurr1 with diminished transcriptional activity

Journal of Neurochemistry ◽

10.1111/j.1471-4159.2005.03458.x ◽

2005 ◽

Vol 95 (5) ◽

pp. 1342-1350 ◽

Cited By ~ 21

Author(s):

Sharon K. Michelhaugh ◽

Henrikas Vaitkevicius ◽

Jun Wang ◽

Mohamad Bouhamdan ◽

Alexys R. Krieg ◽

...

Keyword(s):

Nuclear Receptor ◽

Transcriptional Activity ◽

Dopamine Neurons

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Nuclear receptor HNF4A transrepresses CLOCK:BMAL1 and modulates tissue-specific circadian networks

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1816411115 ◽

2018 ◽

Vol 115 (52) ◽

pp. E12305-E12312 ◽

Cited By ~ 16

Author(s):

Meng Qu ◽

Tomas Duffy ◽

Tsuyoshi Hirota ◽

Steve A. Kay

Keyword(s):

Nuclear Receptor ◽

Metabolic Pathways ◽

Feedback Loop ◽

Transcriptional Activity ◽

Transcript Level ◽

Colon Cells ◽

Tissue Specific ◽

Metabolic Genes ◽

Genome Wide ◽

Circadian Control

Either expression level or transcriptional activity of various nuclear receptors (NRs) have been demonstrated to be under circadian control. With a few exceptions, little is known about the roles of NRs as direct regulators of the circadian circuitry. Here we show that the nuclear receptor HNF4A strongly transrepresses the transcriptional activity of the CLOCK:BMAL1 heterodimer. We define a central role for HNF4A in maintaining cell-autonomous circadian oscillations in a tissue-specific manner in liver and colon cells. Not only transcript level but also genome-wide chromosome binding of HNF4A is rhythmically regulated in the mouse liver. ChIP-seq analyses revealed cooccupancy of HNF4A and CLOCK:BMAL1 at a wide array of metabolic genes involved in lipid, glucose, and amino acid homeostasis. Taken together, we establish that HNF4A defines a feedback loop in tissue-specific mammalian oscillators and demonstrate its recruitment in the circadian regulation of metabolic pathways.

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Identification of a nuclear receptor/coactivator developmental signaling pathway in the nematode parasite Strongyloides stercoralis

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2021864118 ◽

2021 ◽

Vol 118 (8) ◽

pp. e2021864118

Author(s):

Mi Cheong Cheong ◽

Zhu Wang ◽

Tegegn G. Jaleta ◽

Xinshe Li ◽

James B. Lok ◽

...

Keyword(s):

Signaling Pathway ◽

Nuclear Receptor ◽

Transcriptional Activity ◽

Strongyloides Stercoralis ◽

Parasite Development ◽

Binding Motif ◽

Nuclear Receptor Coactivator ◽

Interacting Protein ◽

Third Stage ◽

Infectious Stage

DAF-12 is nematode-specific nuclear receptor that has been proposed to govern development of the infectious stage of parasitic species, including Strongyloides stercoralis. Here, we identified a parasite-specific coactivator, called DAF-12 interacting protein-1 (DIP-1), that is required for DAF-12 ligand-dependent transcriptional activity. DIP-1 is found only in Strongyloides spp. and selectively interacts with DAF-12 through an atypical receptor binding motif. Using CRISPR/Cas9-directed mutagenesis, we demonstrate that DAF-12 is required for the requisite developmental arrest and the ligand-dependent reactivation of infectious S. stercoralis infective third-stage larvae, and that these effects require the DIP-1 coactivator. These studies reveal the existence of a distinct nuclear receptor/coactivator signaling pathway that governs parasite development.

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Type 1 Nuclear Receptor Activity in Breast Cancer: Translating Preclinical Insights to the Clinic

Cancers ◽

10.3390/cancers13194972 ◽

2021 ◽

Vol 13 (19) ◽

pp. 4972

Author(s):

Sanjeev Kumar ◽

Allegra Freelander ◽

Elgene Lim

Keyword(s):

Breast Cancer ◽

Nuclear Receptors ◽

Nuclear Receptor ◽

Patient Outcomes ◽

Estrogen Receptor Α ◽

Receptor Activity ◽

Breast Cancers ◽

Cancer Subtypes ◽

Major Breast

The nuclear receptor (NR) family of transcription factors is intimately associated with the development, progression and treatment of breast cancer. They are used diagnostically and prognostically, and crosstalk between nuclear receptor pathways and growth factor signalling has been demonstrated in all major subtypes of breast cancer. The majority of breast cancers are driven by estrogen receptor α (ER), and anti-estrogenic therapies remain the backbone of treatment, leading to clinically impactful improvements in patient outcomes. This serves as a blueprint for the development of therapies targeting other nuclear receptors. More recently, pivotal findings into modulating the progesterone (PR) and androgen receptors (AR), with accompanying mechanistic insights into NR crosstalk and interactions with other proliferative pathways, have led to clinical trials in all of the major breast cancer subtypes. A growing body of evidence now supports targeting other Type 1 nuclear receptors such as the glucocorticoid receptor (GR), as well as Type 2 NRs such as the vitamin D receptor (VDR). Here, we reviewed the existing preclinical insights into nuclear receptor activity in breast cancer, with a focus on Type 1 NRs. We also discussed the potential to translate these findings into improving patient outcomes.

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

Endocrine Related Cancer ◽

10.1530/erc-16-0319 ◽

2016 ◽

Vol 23 (11) ◽

pp. T85-T108 ◽

Cited By ~ 13

Author(s):

Damien A Leach ◽

Sue M Powell ◽

Charlotte L Bevan

Keyword(s):

Prostate Cancer ◽

Nuclear Receptors ◽

Nuclear Receptor ◽

Current Knowledge ◽

Transcriptional Responses ◽

Number Variation ◽

Genomic Studies ◽

Life Threatening ◽

Castrate Resistant Prostate Cancer ◽

Castrate Resistant

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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