Interrogating the Structural Basis of Nuclear Receptor Activation on Chromatin

探究染色质核受体激活的结构基础

基本信息

批准号：
10538474
负责人：
Timothy Silas Strutzenberg
金额：
$ 6.72万
依托单位：
SALK INSTITUTE FOR BIOLOGICAL STUDIES
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-16 至 2025-09-15
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10538474
关键词：
Adaptive Immune System Area Autoimmune Diseases Binding Biochemical Biochemistry Biological Biological Assay Biological Process Biology Biophysical Process Biophysics Chemicals Chromatin Chromatin Structure Chronic Chronic small plaque psoriasis Clinic Clinical Clinical Trials Complement Complex Consequentialism Cryoelectron Microscopy DNA Data Deuterium Development Disease Drug Targeting Drug usage EP300 gene Electron Microscopy Enzymatic Biochemistry Foundations Gene Activation Gene Expression Gene Expression Regulation Genes Goals Health Histone Acetylation Histones Home Hormones Human Hydrogen Immunity Industrialization Inflammatory Investigation Laboratories Laboratory Study Length Libraries Ligands Light Mass Spectrum Analysis Measurement Mediating Metabolic Diseases Metabolism Methodology Modeling Modification Molecular Molecular Conformation Monitor NCOA3 gene Names Nature Nuclear Receptors Nuclear Structure Nucleosomes Pathology Physiology Play Positioning Attribute Post-Translational Protein Processing Protein Dynamics Proteins Proteomics Receptor Activation Regulation Reproduction Research Resolution Response Elements Role Safety Side Solvents Structure T cell differentiation Techniques Technology Thymus Gland Training Transcriptional Activation Vertebral column Work Writing antimicrobial base biophysical techniques birth control chromatin remodeling crosslink cryogenics design drug development fight against histone modification improved functioning insight member next generation sequencing novel orphan nuclear receptor ROR-gamma preference programs protein complex receptor receptor binding reconstitution recruit side effect structural biology targeted treatment therapeutic target training project transcription factor

项目摘要

PROJECT SUMMARY Nuclear receptors (NRs) are ligand-gated transcription factors that control important biological processes including reproduction, development, metabolism, and immunity. NRs are the target of ~16% of drugs used in the clinic for applications that include birth control, inflammatory disease, and metabolic disorders. NRs are thought to function by altering the dynamics of chromatin to alter expression levels of genes through the recruitment of coregulators, chromatin remodelers, and ultimately through the activation of transcriptional machinery. However, the dynamic nature of NRs and NR complexes has made it challenging to study the molecular mechanisms by which full length proteins and protein assemblies regulate gene expression using traditional structural biology techniques. Despite the central role that NRs play in gene expression activation, our mechanistic understanding of how they regulate target gene programs at the molecular level of chromatin is largely unknown. This is attributed, in part, to the dearth of structural biology data for full-length receptors and the near complete absence of biochemical, biophysical, or structural data on NRs in their functionally relevant chromatin context. Building on established foundations in sponsor’s home laboratory, combined with the applicant’s prior expertise in biochemical and biophysical methods and an intimate understanding of the NR field, this project seeks to determine how a model NR named RORγT assembles with coregulatory proteins SRC3 and p300 to alter chromatin dynamics and post-translational modification status of chromatin, a hallmark of NR target gene activation. The proposed research program has two major goals: (1) to determine how RORγT and coregulatory protein complexes engage with nucleosomes, and (2) to determine how RORγT and coregulator protein complexes modify histones and alter nucleosome dynamics. This will be accomplished using a combination of traditional biochemistry, and state-of-the-art technologies including cryogenic electron microscopy (cryo-EM) and structural proteomics. By studying how these protein complexes assemble and act on nucleosomes, we will gain new insight into how RORγT, and more broadly the NR superfamilty, modulates nucleosome dynamics, modification status, and, ultimately, function in gene regulation. The implications of this work will help us understand how an important drug target functions and will pave the way for future research investigating an important class of drug targets.

项目摘要核受体（NRS）是控制重要生物过程的配体门控转录因子包括繁殖，发展，代谢和免疫力。 NRS是约16％用于使用的药物的靶标包括节育，炎症性疾病和代谢性疾病的应用诊所。 NRS是认为通过改变染色质的动力学来改变基因的表达水平，从而通过募集核控剂，染色质重塑剂，并最终通过转录激活机械。但是，NRS和NR复合物的动态性质使研究全长蛋白质和蛋白质组件使用的分子机制使用该机制调节基因表达传统的结构生物学技术。尽管NRS在基因表达激活中起着核心作用，但我们对它们如何调节染色质分子水平的靶基因程序的机械理解是在很大程度上未知。这部分归因于全长受体的结构生物学数据和在功能相关的NRS上几乎完全没有生化，生物物理或结构数据染色质上下文。建立在赞助商的家庭实验室中的既定基础，结合申请人先前在生化和生物物理方法方面的专业知识以及对NR领域的深入了解，该项目旨在确定NR命名RORγT的模型如何用核心调节蛋白SRC3组装和p300以改变染色质动力学和染色质的翻译后修饰状态，NR的标志靶基因激活。拟议的研究计划有两个主要目标：（1）确定RORγT和核调节蛋白复合物与核小体接合，（2）确定RORγT和COREGULATOR如何蛋白质复合物可修饰组蛋白并改变核小体动力学。这将使用传统生物化学和最先进的技术（包括低温电子）的结合显微镜（冷冻EM）和结构蛋白质组学。通过研究这些蛋白质复合物如何组装和作用在核小体上，我们将获得有关RORγT和更广泛的NR超家族的新见解。核小体动力学，修饰状态以及最终在基因调节中的功能。这个含义工作将帮助我们了解重要的药物目标如何运作，并为将来的研究铺平道路研究一类重要的药物靶标。