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

• 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
• 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

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.

项目概要 核受体 (NR) 是控制重要生物过程的配体门控转录因子 包括生殖、发育、新陈代谢和免疫。 NR 是约 16% 的药物的目标 该诊所的应用包括节育、炎症性疾病和代谢紊乱。 NR 是 人们认为通过改变染色质的动态来改变基因的表达水平 招募辅助调节因子、染色质重塑因子，并最终通过转录激活 机械。然而，NR 和 NR 复合物的动态性质使得研究它们变得具有挑战性。 全长蛋白质和蛋白质组装体调节基因表达的分子机制 传统的结构生物学技术。尽管 NR 在基因表达激活中发挥着核心作用，但我们的 对它们如何在染色质分子水平上调节靶基因程序的机制理解是 很大程度上不为人所知。这部分归因于缺乏全长受体的结构生物学数据和 NR 的功能相关的生化、生物物理或结构数据几乎完全缺失 染色质背景。以申办者家庭实验室的既定基础为基础，结合 申请人先前在生物化学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， 该项目旨在确定名为 RORγT 的模型 NR 如何与辅助调节蛋白 SRC3 组装 和 p300 改变染色质动力学和染色质翻译后修饰状态，这是 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 核心调节蛋白复合物与核小体结合，以及 (2) 确定 RORγT 和核心调节因子如何 蛋白质复合物修饰组蛋白并改变核小体动力学。这将通过使用 传统生物化学与包括低温电子在内的最先进技术的结合 显微镜（冷冻电镜）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发挥作用 在核小体上，我们将获得关于 RORγT 以及更广泛的 NR 超家族如何调节的新见解 核小体动力学、修饰状态以及最终的基因调控功能。这的影响 这项工作将帮助我们了解重要的药物靶标如何发挥作用，并为未来的研究铺平道路 研究一类重要的药物靶点。