Uncovering the function of histone variant H2BE in neurons

揭示组蛋白变体 H2BE 在神经元中的功能

• 批准号: 10610728
• 负责人: Emily Ruth Feierman Hyatt
• 金额: $ 3.51万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10610728
• 关键词: ATAC-seq; Address; Affect; Animal Behavior; Antibodies; Atomic Force Microscopy; Behavior; Behavioral; Binding; Biochemical; Biology; Brain; Brain region; Brain-Derived Neurotrophic Factor; Cell physiology; Cells; ChIP-seq; Chromatin; Chromatin Fiber; Chromatin Structure; Cognition; Cognitive; Complex; DNA Packaging; Data; Data Set; Environment; Epigenetic Process; Exposure to; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Genomic Segment; Genomics; Goals; Heterochromatin; Histone H1; Histones; Immediate-Early Genes; Knock-out; Knockout Mice; Learning; Link; Longevity; Mass Spectrum Analysis; Memory; Memory Disorders; Molecular; Mus; Neurons; Nucleosomes; Olfactory Epithelium; Olfactory Pathways; Output; Pathway interactions; Phenotype; Post-Translational Protein Processing; Proteins; Regulation; Research; Role; Short-Term Memory; Signal Transduction; Sorting; Stimulus; Synapses; Techniques; Testing; Therapeutic; Tissues; Transcriptional Regulation; Translating; Treatment Factor; Variant; Work; behavior influence; behavior test; behavioral response; behavioral study; design; environmental enrichment for laboratory animals; epigenetic regulation; extracellular; fear memory; flexibility; gene repression; genome-wide; insight; interest; long term memory; mouse model; nervous system disorder; novel; postmitotic; response; transcriptome sequencing; whole genome

Project Summary The goal of this proposal is to elucidate the molecular and behavioral function of histone variant H2BE in neurons. Histone variants, which are encoded by separate genes, can substitute for the canonical histone proteins (H2A, H2B, H3, and H4) and are involved in regulation of many cellular processes and gene expression. The histone variant H2BE was discovered in the mouse main olfactory epithelium, where it affects olfactory neuron function and longevity. While H2BE was previously thought to be exclusively expressed in the olfactory system, our lab developed a highly specific antibody against H2BE and demonstrated that H2BE is present throughout the brain. However, despite the importance of histone variants in controlling neuronal function, to date, H2BE remains unstudied outside of the olfactory system. Here, I propose to determine how H2BE alters chromatin structure, neuronal gene expression, and animal behavior. I hypothesize that H2BE decreases binding of linker histone H1, controls expression of activity-dependent genes, and is necessary for cognitive flexibility, spatial learning, and fear memory. To test my hypothesis, I will combine genome-wide sequencing, mouse models, and animal behavior with molecular and biochemical techniques from the chromatin biology field. In Aim 1, I will determine how H2BE expression alters chromatin structure. I will use ChIP-sequencing to define the genomic localization of H2BE at baseline and in response to external signals. My preliminary data demonstrates that H2BE promotes an open chromatin configuration and decreases binding of heterochromatin-associated proteins. Therefore, I will examine how H2BE affects the composition of the chromatin fiber. Specifically, I will use ChIP-seq to determine how H2BE incorporation affects localization of linker histone H1. Aim 2 addresses the role of H2BE in neuronal gene expression and mouse behavior. First, I will test the effects of H2BE knockout on RNA-sequencing of neurons with and without external stimulation. Second, I will perform a battery of behavioral tests using H2BE WT and KO mice designed to determine the specific brain regions most affected by H2BE loss and to fully characterize the role of H2BE in cognition. The work proposed here will reveal how histone variant H2BE contributes to the complex chromatin environment in the brain. This discovery is critical to understanding how neurons use environmental signals to control transcription and ultimately govern behavior. In developing a more complete understanding of the chromatin landscape in neurons, we will also gain insight into potential therapeutics for the treatment of the many neurological disorders that are linked to disruption of epigenetic regulation in the brain.

项目概要 该提案的目标是阐明组蛋白变体 H2BE 在神经元中的分子和行为功能。 由不同基因编码的组蛋白变体可以替代典型的组蛋白（H2A、 H2B、H3 和 H4）并参与许多细胞过程和基因表达的调节。组蛋白 在小鼠主嗅上皮中发现了变异 H2BE，它影响嗅觉神经元功能 和长寿。虽然 H2BE 以前被认为只在嗅觉系统中表达，但我们的实验室 开发了一种针对 H2BE 的高度特异性抗体，并证明 H2BE 存在于整个大脑中。 然而，尽管组蛋白变异在控制神经元功能中很重要，但迄今为止，H2BE 仍然 在嗅觉系统之外尚未进行研究。在这里，我建议确定 H2BE 如何改变染色质结构， 神经元基因表达和动物行为。我假设 H2BE 减少了接头组蛋白的结合 H1，控制活动依赖性基因的表达，对于认知灵活性、空间学习、 和恐惧记忆。为了验证我的假设，我将结合全基因组测序、小鼠模型和动物模型 染色质生物学领域的分子和生化技术的行为。在目标 1 中，我将确定 H2BE 表达如何改变染色质结构。我将使用 ChIP 测序来定义基因组定位 H2BE 的基线和对外部信号的响应。我的初步数据表明 H2BE 促进 开放的染色质构型并减少异染色质相关蛋白的结合。因此，我将 检查 H2BE 如何影响染色质纤维的组成。具体来说，我将使用 ChIP-seq 来确定 H2BE 掺入如何影响接头组蛋白 H1 的定位。目标 2 阐述了 H2BE 在神经元中的作用 基因表达和小鼠行为。首先，我将测试 H2BE 敲除对 RNA 测序的影响 有或没有外部刺激的神经元。其次，我将使用 H2BE 执行一系列行为测试 WT 和 KO 小鼠旨在确定受 H2BE 损失影响最大的特定大脑区域，并充分了解 描述 H2BE 在认知中的作用。这里提出的工作将揭示组蛋白变体 H2BE 如何 有助于大脑中复杂的染色质环境。这一发现对于理解如何 神经元利用环境信号来控制转录并最终控制行为。在开发更多 在完全了解神经元染色质景观的同时，我们还将深入了解潜在的 用于治疗许多与表观遗传破坏有关的神经系统疾病的疗法 大脑中的调节。