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Role of Allostery in CAR Transactivation

变构在 CAR 反式激活中的作用

基本信息

批准号：
9305482
负责人：
ELIAS J FERNANDEZ
金额：
$ 45.3万
依托单位：
UNIVERSITY OF TENNESSEE KNOXVILLE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-06-01 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9305482
关键词：
Address Bilirubin Binding Binding Sites Biological Biological Assay Biological Process Biology Biophysics Cells Complex Crystallography DNA DNA Binding DNA Sequence Data Development Disease Distant Endocrine Family Gene Expression Gene Expression Profile Genetic Transcription Genomic approach Goals Hormonal Human In Vitro Individual Knowledge Laboratories Ligand Binding Ligands Link Liver neoplasms Mediating Metabolic Minor Modeling Molecular Molecular Conformation Nuclear Receptors Outcome Pathway interactions Pattern Pharmaceutical Preparations Pharmacologic Substance Pharmacology Physiology Play Proteins Psychological Techniques RXR Recruitment Activity Reporter Genes Research Personnel Response Elements Role Signaling Molecule Site Specificity Stimulus Structure Students System Techniques Testing Theoretical Studies Toxin Transactivation X-Ray Crystallography Xenobiotics base biophysical analysis biophysical properties biophysical techniques chromatin immunoprecipitation chromatin modification cofactor constitutive androstane receptor experimental study extracellular genome-wide in vivo interest knock-down lipophilicity liver development mouse model novel receptor receptor binding receptor function response scaffold skills small hairpin RNA small molecule three dimensional structure transcription factor transcriptome sequencing

项目摘要

Structure-Function Studies on CAR The primary goal of this project is to determine the mechanisms of action of the constitutive androstane receptor (CAR). CAR plays key roles in the clearance of xenobiotics and endogenous toxins such as bilirubin. CAR belongs to the nuclear receptor family of transcription factor proteins that mediate endocrine function and play critical roles in development and physiology and pharmacology. The functional activity of nuclear receptors is regulated by small molecular hormonal and synthetic molecules. CAR is regulated by small-molecule signals. Earlier, we produced preliminary evidence utilizing multiple biophysical and cell-based assays to show how distinct allosteric pathways can fine-tune the activity of nuclear receptors. Our goal here is to identify and distinguish between the different allosteric pathways initiated by ligands, DNA and cofactors such as co- activators and co-repressors. We will use a combination of crystallography and biophysical techniques to determine the local and distant conformational changes that occur upon ligand binding. With Dr. Tongye Shen we will use theoretical techniques to predict allosteric pathways that can be further characterized by experimental techniques. We will also use genomic approaches in colla boration with Dr. Rachel Patton McCord to determine the role of allostery, in vivo. The ultimate goal of this study is to develop our understanding of CAR and nuclear receptors. Malfunctioning nuclear receptors are associated with several disease states, and CAR activity has been associated with the development of liver tumors in mouse models. In summary, we will address the following questions: (i) What conformational pathways link ligand, DNA and coregulatory molecules? (ii) How do these pathways regulate coactivator and DNA recognition? (iii) What is the genome-scale impact of allostery in CAR? Thus, a detailed understanding of these proteins, and CAR, will enable us to develop efficient therapies in the long term.

轿车的结构-功能研究这个项目的主要目标是确定构成雄烷受体(CAR)。汽车在清理环境污染中起着关键作用外源物质和内源性毒素，如胆红素。汽车属于核能介导内分泌功能的转录因子受体家族蛋白并在发育、生理学和药理学中发挥关键作用。这个核受体的功能活性受小分子激素调节和合成分子。CAR受到小分子信号的调节。此前，我们利用多种生物物理和基于细胞的方法产生了初步证据显示不同的变构途径可以微调活性的化验核受体。我们在这里的目标是识别和区分不同的变构途径由配体、DNA和辅因子等共同启动。激活剂和共抑制物。我们将结合使用结晶学和确定局部和远程构象变化的生物物理技术发生在配基结合时。与沈同业博士一起，我们将用理论预测变构途径的技术，可以进一步表征实验技术。我们还将在Colla Boration中使用基因组方法与雷切尔·巴顿·麦考德博士一起确定变构在体内的作用。这个这项研究的最终目的是加深我们对汽车和核能的理解感受器。核受体功能障碍与几种疾病有关状态，CAR活动与肝脏的发育有关小鼠模型中的肿瘤。综上所述，我们将解决以下问题：(I) 连接配体、DNA和共调节分子的构象途径是什么？(Ii) 这些途径是如何调节辅活化子和DNA识别的？(Iii)何谓变构在CAR中的基因组规模影响？因此，详细了解这些蛋白质和CAR将使我们能够长期开发有效的治疗方法学期。