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1.
Mol Metab ; 84: 101938, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38631478

RESUMO

OBJECTIVE: The peroxisome proliferator-activated receptor α (PPARα) is a transcription factor driving target genes involved in fatty acid ß-oxidation. To what extent various PPARα interacting proteins may assist its function as a transcription factor is incompletely understood. An ORFeome-wide unbiased mammalian protein-protein interaction trap (MAPPIT) using PPARα as bait revealed a PPARα-ligand-dependent interaction with the orphan nuclear receptor estrogen-related receptor α (ERRα). The goal of this study was to characterize the nature of the interaction in depth and to explore whether it was of physiological relevance. METHODS: We used orthogonal protein-protein interaction assays and pharmacological inhibitors of ERRα in various systems to confirm a functional interaction and study the impact of crosstalk mechanisms. To characterize the interaction surfaces and contact points we applied a random mutagenesis screen and structural overlays. We pinpointed the extent of reciprocal ligand effects of both nuclear receptors via coregulator peptide recruitment assays. On PPARα targets revealed from a genome-wide transcriptome analysis, we performed an ERRα chromatin immunoprecipitation analysis on both fast and fed mouse livers. RESULTS: Random mutagenesis scanning of PPARα's ligand-binding domain and coregulator profiling experiments supported the involvement of (a) bridging coregulator(s), while recapitulation of the interaction in vitro indicated the possibility of a trimeric interaction with RXRα. The PPARα·ERRα interaction depends on 3 C-terminal residues within helix 12 of ERRα and is strengthened by both PGC1α and serum deprivation. Pharmacological inhibition of ERRα decreased the interaction of ERRα to ligand-activated PPARα and revealed a transcriptome in line with enhanced mRNA expression of prototypical PPARα target genes, suggesting a role for ERRα as a transcriptional repressor. Strikingly, on other PPARα targets, including the isolated PDK4 enhancer, ERRα behaved oppositely. Chromatin immunoprecipitation analyses demonstrate a PPARα ligand-dependent ERRα recruitment onto chromatin at PPARα-binding regions, which is lost following ERRα inhibition in fed mouse livers. CONCLUSIONS: Our data support the coexistence of multiple layers of transcriptional crosstalk mechanisms between PPARα and ERRα, which may serve to finetune the activity of PPARα as a nutrient-sensing transcription factor.


Assuntos
Receptor ERRalfa Relacionado ao Estrogênio , PPAR alfa , Receptores de Estrogênio , PPAR alfa/metabolismo , PPAR alfa/genética , Animais , Camundongos , Receptores de Estrogênio/metabolismo , Receptores de Estrogênio/genética , Humanos , Regulação da Expressão Gênica , Células HEK293 , Masculino , Camundongos Endogâmicos C57BL , Ligação Proteica , Fígado/metabolismo
2.
Nat Rev Endocrinol ; 16(7): 363-377, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32303708

RESUMO

Nuclear receptor crosstalk can be defined as the interplay between different nuclear receptors or between their overlapping signalling pathways. A subset of nuclear receptors (such as PPARs and RARs) engage in the formation of well-characterized 'typical' heterodimers with RXR. 'Atypical' heterodimers (such as GR with PPARs, or PPAR with ERR) might form a novel class of physical complexes that might be more transient in nature. These heterodimers might harbour strong transcriptional flexibility, with no strict need for DNA binding of both partners. Direct crosstalk could stem from a pairwise physical association between atypical nuclear receptor heterodimers, either via pre-existing interaction pairs or via interactions that are newly induced with small molecules; such crosstalk might constitute an uncharted space to target nuclear receptor physiological and/or pathophysiological actions. In this Review, we discuss the emerging aspects of crosstalk in the nuclear receptor field and present various mechanistic crosstalk modes with examples that support applicability of the atypical heterodimer concept. Stabilization or disruption, in a context-dependent or cell type-dependent manner, of these more transient heterodimers is expected to fuel unprecedented translational approaches to yield novel therapeutic agents to treat major human diseases with higher precision.


Assuntos
Receptor Cross-Talk/fisiologia , Receptores Citoplasmáticos e Nucleares/fisiologia , Terapias em Estudo/métodos , Animais , Humanos , Ligação Proteica/fisiologia , Multimerização Proteica/fisiologia , Receptores Citoplasmáticos e Nucleares/metabolismo , Transdução de Sinais/fisiologia , Terapias em Estudo/tendências , Fatores de Transcrição/metabolismo , Fatores de Transcrição/fisiologia
3.
Endocr Rev ; 39(5): 760-802, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30020428

RESUMO

Peroxisome proliferator-activated receptor α (PPARα) is a nuclear receptor of clinical interest as a drug target in various metabolic disorders. PPARα also exhibits marked anti-inflammatory capacities. The first-generation PPARα agonists, the fibrates, have however been hampered by drug-drug interaction issues, statin drop-in, and ill-designed cardiovascular intervention trials. Notwithstanding, understanding the molecular mechanisms by which PPARα works will enable control of its activities as a drug target for metabolic diseases with an underlying inflammatory component. Given its role in reshaping the immune system, the full potential of this nuclear receptor subtype as a versatile drug target with high plasticity becomes increasingly clear, and a novel generation of agonists may pave the way for novel fields of applications.


Assuntos
Inflamação/metabolismo , Metabolismo dos Lipídeos/fisiologia , PPAR alfa/metabolismo , Animais , Humanos , Fígado/metabolismo
4.
J Clin Invest ; 127(4): 1136-1145, 2017 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-28319043

RESUMO

Glucocorticoids (GCs; referred to clinically as corticosteroids) are steroid hormones with potent anti-inflammatory and immune modulatory profiles. Depending on the context, these hormones can also mediate pro-inflammatory activities, thereby serving as primers of the immune system. Their target receptor, the GC receptor (GR), is a multi-tasking transcription factor, changing its role and function depending on cellular and organismal needs. To get a clearer idea of how to improve the safety profile of GCs, recent studies have investigated the complex mechanisms underlying GR functions. One of the key findings includes both pro- and anti-inflammatory roles of GR, and a future challenge will be to understand how such paradoxical findings can be reconciled and how GR ultimately shifts the balance to a net anti-inflammatory profile. As such, there is consensus that GR deserves a second life as a drug target, with either refined classic GCs or a novel generation of nonsteroidal GR-targeting molecules, to meet the increasing clinical needs of today to treat inflammation and cancer.


Assuntos
Analgésicos/uso terapêutico , Sistemas de Liberação de Medicamentos , Glucocorticoides/imunologia , Proteínas de Neoplasias/imunologia , Receptores de Glucocorticoides/imunologia , Animais , Humanos , Inflamação/imunologia , Inflamação/patologia , Inflamação/terapia , Neoplasias/tratamento farmacológico , Neoplasias/imunologia , Neoplasias/patologia
5.
Methods Mol Biol ; 1204: 83-94, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25182763

RESUMO

The Microarray Assay for Realtime Coregulator-Nuclear receptor Interaction (MARCoNI) technology allows the identification of nuclear receptor-coregulator interactions via flow-through microarrays. As such, differences in the coregulator profile between distinct nuclear receptors or of a single receptor in agonist or antagonist mode can be investigated, even in a single run. In this chapter, the method how to perform these peptide microarrays with cell lysates containing the overexpressed glucocorticoid receptor is described, as well as the influence of assay parameters, variations to the protocol, and data analysis.


Assuntos
Análise Serial de Proteínas/métodos , Receptores de Glucocorticoides/metabolismo , Animais , Células HEK293 , Humanos , Software
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