Nuclear receptor signaling最新文献_第7页

An improved high throughput protein-protein interaction assay for nuclear hormone receptors. 核激素受体的一种改进的高通量蛋白相互作用测定方法。

Nuclear receptor signaling Pub Date : 2007-03-09 DOI: 10.1621/nrs.05002

Michael L Goodson, Behnom Farboud, Martin L Privalsky

{"title":"An improved high throughput protein-protein interaction assay for nuclear hormone receptors.","authors":"Michael L Goodson, Behnom Farboud, Martin L Privalsky","doi":"10.1621/nrs.05002","DOIUrl":"https://doi.org/10.1621/nrs.05002","url":null,"abstract":"The Glutathione-S-Transferase (GST) \"pulldown\" assay has been used extensively to assay protein interactions in vitro. This methodology has been especially useful for investigating the interactions of nuclear hormone receptors with a wide variety of their interacting partners and coregulatory proteins. Unfortunately, the original GST-pulldown technique relies on multiple binding, washing and elution steps performed in individual microfuge tubes, and requires repeated centrifugation, aspiration, and suspension steps. This type of batch processing creates a significant liquid handling bottleneck, limiting the number of sample points that can be incorporated into one experiment and producing inherently less efficient washing and elution than would a flow-through methodology. In this manuscript, we describe the adaptation of this GST-pulldown assay to a 96-well filter plate format. The use of a multi-well filter plate makes it possible to assay more samples in significantly less time using less reagents and more efficient sample processing than does the traditional single tube assay.","PeriodicalId":87415,"journal":{"name":"Nuclear receptor signaling","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2007-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1621/nrs.05002","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26691249","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 19

Natural disordered sequences in the amino terminal domain of nuclear receptors: lessons from the androgen and glucocorticoid receptors. 核受体氨基末端区域的自然无序序列:来自雄激素和糖皮质激素受体的教训。

Nuclear receptor signaling Pub Date : 2007-03-09 DOI: 10.1621/nrs.05001

Iain J McEwan, Derek Lavery, Katharina Fischer, Kate Watt

引用次数: 12

Visualizing the action of steroid hormone receptors in living cells. 可视化活细胞中类固醇激素受体的作用。

Nuclear receptor signaling Pub Date : 2007-03-09 DOI: 10.1621/nrs.05003

Alexander Griekspoor, Wilbert Zwart, Jacques Neefjes, Rob Michalides

引用次数: 89

Corepressor/coactivator paradox: potential constitutive coactivation by corepressor splice variants. 核心抑制器/协同激活器悖论：核心抑制器剪接变体的潜在构成性协同激活。

Nuclear receptor signaling Pub Date : 2006-10-30 DOI: 10.1621/nrs.04022

Xianwang Meng, Vishnuka D Arulsundaram, Ahmed F Yousef, Paul Webb, John D Baxter, Joe S Mymryk, Paul G Walfish

{"title":"Corepressor/coactivator paradox: potential constitutive coactivation by corepressor splice variants.","authors":"Xianwang Meng, Vishnuka D Arulsundaram, Ahmed F Yousef, Paul Webb, John D Baxter, Joe S Mymryk, Paul G Walfish","doi":"10.1621/nrs.04022","DOIUrl":"10.1621/nrs.04022","url":null,"abstract":"The functional consequences of the interaction of transcriptional coregulators with the human thyroid hormone receptor (TR) in mammalian cells are complex. We have used the yeast, Saccharomyces cerevisiae, which lack endogenous nuclear receptors (NRs) and NR coregulators, as a model to decipher mechanisms regulating transcriptional activation by TR. In effect, this system allows the reconstitution of TR mediated transcription complexes by the expression of specific combinations of mammalian proteins in yeast. In this yeast system, human adenovirus 5 early region 1A (E1A), a natural N-CoR splice variant (N-CoR(I)) or an artificial N-CoR truncation (N-CoR(C)) coactivate unliganded TRs and these effects are inhibited by thyroid hormone (TH). E1A contains a short peptide sequence that resembles known corepressor-NR interaction motifs (CoRNR box motif, CBM), and this motif is required for TR binding and coactivation. N-CoR(I) and N-CoR(C) contain three CBMs, but only the C-terminal CBM1 is critical for coactivation. These observations in a yeast model system suggest that E1A and N-CoR(I) are naturally occurring TR coactivators that bind in the typical corepressor mode. These findings also raise the possibility that alternative splicing events which form corepressor proteins containing only C-terminal CBM motifs could represent a novel mechanism in mammalian cells for regulating constitutive transcriptional activation by TRs.","PeriodicalId":87415,"journal":{"name":"Nuclear receptor signaling","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2006-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1630687/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26409184","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The nuclear receptor transcriptional coregulator RIP140. 核受体转录共调节因子RIP140。

Nuclear receptor signaling Pub Date : 2006-10-30 DOI: 10.1621/nrs.04024

Patrick Augereau, Eric Badia, Sophie Carascossa, Audrey Castet, Samuel Fritsch, Pierre-Olivier Harmand, Stéphan Jalaguier, Vincent Cavaillès

{"title":"The nuclear receptor transcriptional coregulator RIP140.","authors":"Patrick Augereau, Eric Badia, Sophie Carascossa, Audrey Castet, Samuel Fritsch, Pierre-Olivier Harmand, Stéphan Jalaguier, Vincent Cavaillès","doi":"10.1621/nrs.04024","DOIUrl":"https://doi.org/10.1621/nrs.04024","url":null,"abstract":"The nuclear receptor superfamily comprises ligand-regulated transcription factors that control various developmental and physiological pathways. These receptors share a common modular structure and regulate gene expression through the recruitment of a large set of coregulatory proteins. These transcription cofactors regulate, either positively or negatively, chromatin structure and transcription initiation. One of the first proteins to be identified as a hormone-recruited cofactor was RIP140. Despite its recruitment by agonist-liganded receptors, RIP140 exhibits a strong transcriptional repressive activity which involves several inhibitory domains and different effectors. Interestingly, the RIP140 gene, located on chromosome 21 in humans, is finely regulated at the transcriptional level by various nuclear receptors. In addition, the protein undergoes several post-translational modifications which control its repressive activity. Finally, experiments performed in mice devoid of the RIP140 gene indicate that this transcriptional cofactor is essential for female fertility and energy homeostasis. RIP140 therefore appears to be an important modulator of nuclear receptor activity which could play major roles in physiological processes and hormone-dependent diseases.","PeriodicalId":87415,"journal":{"name":"Nuclear receptor signaling","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2006-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1621/nrs.04024","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26350603","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 57

A critique of Women's Health Initiative Studies (2002-2006). 《妇女健康倡议研究评论》(2002-2006)。

Nuclear receptor signaling Pub Date : 2006-10-30 DOI: 10.1621/nrs.04023

James H Clark

引用次数: 50

Measuring ligand-dependent and ligand-independent interactions between nuclear receptors and associated proteins using Bioluminescence Resonance Energy Transfer (BRET). 利用生物发光共振能量转移(BRET)测量核受体与相关蛋白之间的配体依赖性和非配体依赖性相互作用。

Nuclear receptor signaling Pub Date : 2006-07-26 DOI: 10.1621/nrs.04021

Kristen L Koterba, Brian G Rowan

{"title":"Measuring ligand-dependent and ligand-independent interactions between nuclear receptors and associated proteins using Bioluminescence Resonance Energy Transfer (BRET).","authors":"Kristen L Koterba, Brian G Rowan","doi":"10.1621/nrs.04021","DOIUrl":"https://doi.org/10.1621/nrs.04021","url":null,"abstract":"Bioluminescent resonance energy transfer (BRET2) is a recently developed technology for the measurement of protein-protein interactions in a live, cell-based system. BRET2 is characterized by the efficient transfer of excited energy between a bioluminescent donor molecule (Renilla luciferase) and a fluorescent acceptor molecule (a mutant of Green Fluorescent Protein (GFP2). The BRET2 assay offers advantages over fluorescence resonance energy transfer (FRET) because it does not require an external light source thereby eliminating problems of photobleaching and autoflourescence. The absence of contamination by light results in low background that permits detection of very small changes in the BRET2 signal. BRET2 is dependent on the orientation and distance between two fusion proteins and therefore requires extensive preliminary standardization experiments to conclude a positive BRET2 signal independent of variations in protein titrations and arrangement in tertiary structures. Estrogen receptor (ER) signaling is modulated by steroid receptor coactivator 1 (SRC-1). To establish BRET2 in a ligand inducible system we used SRC-1 as the donor moiety and ER as the acceptor moiety. Expression and functionality of the fusion proteins were assessed by transient transfection in HEK-293 cells followed by Western blot analysis and measurement of ER-dependent reporter gene activity. These preliminary determinations are required prior to measuring nuclear receptor protein-protein interactions by BRET2. This article describes in detail the BRET2 methodology for measuring interaction between full-length ER and coregulator proteins in real-time, in an in vivo environment.","PeriodicalId":87415,"journal":{"name":"Nuclear receptor signaling","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2006-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1621/nrs.04021","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26287884","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 23

A scoring system for the follow up study of nuclear receptor coactivator complexes. 核受体辅激活物后续研究的评分系统。

Nuclear receptor signaling Pub Date : 2006-01-01 Epub Date: 2006-07-07 DOI: 10.1621/nrs.04014

Sang Jun Han, Sung Yun Jung, Anna Malovannaya, Taeil Kim, Rainer B Lanz, Jun Qin, Bert W O'Malley

{"title":"A scoring system for the follow up study of nuclear receptor coactivator complexes.","authors":"Sang Jun Han, Sung Yun Jung, Anna Malovannaya, Taeil Kim, Rainer B Lanz, Jun Qin, Bert W O'Malley","doi":"10.1621/nrs.04014","DOIUrl":"https://doi.org/10.1621/nrs.04014","url":null,"abstract":"We have systematically isolated a variety of coactivator complexes from HeLa S3 cells using proteomic approaches. In the present report, we have evaluated twelve coactivator complexes involved in nuclear receptor-dependent gene transcription that have been purified by using an immunoprecipitation method. The twelve purified coactivator complexes are SRC-1, SRC-2, SRC-3, CBP, p300, CAPER, E6-AP, ASC-1, CoREST, CRSP3, CRSP2, and CDK7 containing complexes. We have identified 153 protein components associated with these coactivator complexes using mass spectrometry. In order to systematically characterize the functional roles for these components in nuclear receptor-dependent gene transcription and their investigative potential, we have developed a scoring system. This scoring system is comprised of biological and experimental parameters. The biological evaluation considers aspects such as intrinsic enzymatic activity of a protein component, cellular signaling processes in which protein components may be involved, associations with human disease, specific protein motifs, and the known biological roles of other interacting partners of the identified protein. In the experimental evaluation, we include parameters, such as the availability of research materials for the functional study of the identified protein component; such as full-length cDNA clones, antibodies, and commercially available knock-out embryonic stem (ES) cells. Each scoring parameter has been assigned an arbitrary number of points according to perceived relative importance. On the basis of this scoring system, we prioritized each of the protein components in terms of the likelihood of their importance for coactivator complex networking in nuclear receptor-dependent gene transcription.","PeriodicalId":87415,"journal":{"name":"Nuclear receptor signaling","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2006-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1621/nrs.04014","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26160488","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 8

Regulation of progesterone receptor signaling by BRCA1 in mammary cancer. BRCA1在乳腺癌中对孕激素受体信号的调控。

Nuclear receptor signaling Pub Date : 2006-01-01 Epub Date: 2006-04-28 DOI: 10.1621/nrs.04006

Pragati Katiyar, Yongxian Ma, Saijun Fan, Richard G Pestell, Priscilla A Furth, Eliot M Rosen

引用次数: 29

Integrating nuclear receptor mobility in models of gene regulation. 整合核受体迁移的基因调控模型。

Nuclear receptor signaling Pub Date : 2006-01-01 Epub Date: 2006-04-28 DOI: 10.1621/nrs.04010

Laurent Gelman, Jerome N Feige, Cicerone Tudor, Yves Engelborghs, Walter Wahli, Beatrice Desvergne

引用次数: 12