Lysine methylation at chromatin and cellular responses to stress

染色质赖氨酸甲基化和细胞对应激的反应

基本信息

批准号：
10197952
负责人：
Erin Green
金额：
$ 29.1万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE COUNTY
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：
2017-07-15 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10197952
关键词：
Address Aging Binding Biochemical Biological Assay Cell Maintenance Cell Survival Cells Cellular Stress ChIP-seq Chromatin Chromatin Remodeling Factor Chromatin Structure Cues DNA Packaging Data Defect Disease Environment Environmental Risk Factor Enzymes Epigenetic Process Foundations Gene Expression Gene Expression Profile Gene Expression Regulation Genes Genetic Genome Genome Stability Genomics Grant Histone H2A Histones Homeostasis Human In Vitro Investigation Knowledge Link Lysine Malignant Neoplasms Mass Spectrum Analysis Mediating Methylation Methyltransferase Modification Molecular Molecular Analysis Molecular Target Mutation Neurodevelopmental Disorder Oncogenic Orthologous Gene Oxidative Stress PHD Finger Pathologic Pathologic Processes Pathology Pathway interactions Pattern Play Positioning Attribute Post-Translational Protein Processing Proteins Proteomics Reader Regulation Research Resolution Role SET Domain Saccharomyces cerevisiae Proteins Series Signal Transduction Site Stimulus Stress Stress Response Signaling Stress Tests Structure Substrate Specificity System Tertiary Protein Structure Testing Work Yeasts base biological adaptation to stress environmental stressor epigenetic regulation epigenomics experimental study genetic analysis histone methylation histone methyltransferase histone modification human disease insight mutant novel preference programs protein protein interaction response sensor stem cells transcriptome sequencing tumorigenesis

项目摘要

Project Summary Cell survival in the presence of fluctuating environmental signals is critically dependent on rapid changes in gene expression. Chromatin-modifying enzymes are key regulators of genome reprogramming during stress, and aberrant regulation or mutation of these enzymes results in disrupted gene expression programs and inappropriate responses to cellular stress. Such consequences contribute to pathological processes including oncogenesis and aging of human cells. Despite critical roles for chromatin modifiers in these pathways, there are still substantial gaps in our knowledge regarding novel sites of histone modification and their effects on genome regulation, particularly in the presence of diverse stresses encountered in the environment. Our preliminary work has uncovered that the Saccharomyces cerevisiae protein Set4, a potential ortholog of the human protein MLL5, is important for cell survival in oxidative stress and that it is an active histone methyltransferase. The central hypothesis of our work is that Set4 is a stress- regulated methyltransferase that activates a defined gene expression program in response to stress through its lysine methylation activity. Three specific aims are proposed. In Aim I, we will define the substrate specificity of Set4 both in vitro and in cells using biochemical and mass spectrometry based approaches. We will test whether Set4 primarily targets histones, or if it also has additional, non-histone methyl-lysine substrates that contribute to the oxidative stress response. In Aim II, molecular and genetic analysis will be used to determine pathways that regulate Set4 itself in response to oxidative, and other, stresses. Mechanisms by which Set4 controls gene expression will be elucidated through RNA-sequencing analysis and chIP- sequencing of Set4 and its cognate methyl mark in a series of mutants under stress. Aim III will test the hypothesis that the PHD finger of Set4 is required for Set4-depdent stress responses by promoting its localization to chromatin. Biochemical and proteomic assays will determine the histone or non-histone binding partner of the PHD finger of Set4, and targeted molecular and genomic experiments will assess the role of the PHD finger in the localization and activity of Set4 at chromatin. These research aims will provide substantial insight in to the function of a novel epigenetic modifier that we expect to be applicable to its potential human ortholog MLL5, which has been implicated in stem cell maintenance, tumorigenesis and neurodevelopmental disorders. Furthermore, this work will uncover new links between environmental stress and chromatin-based regulation of gene expression, which will be critical to our understanding of how misregulation of the genome by aberrant stress signaling contributes to human disease.

项目摘要在存在波动环境信号的情况下，细胞存活至关重要取决于快速基因表达的变化。染色质改良酶是基因组的关键调节剂在压力期间重新编程以及这些酶的异常调节或突变导致破坏基因表达程序和对细胞应激的不适当反应。这样的后果有助于病理过程，包括人类的肿瘤发生和衰老细胞。尽管染色质修饰剂在这些途径中的关键作用，但仍然存在很大的我们关于组蛋白修饰的新颖地点的知识及其对基因组的影响的差距调节，特别是在环境中遇到的各种压力的情况下。我们的初步工作发现了酿酒酵母蛋白set4的潜力人蛋白Mll5的直系同源物对于氧化应激中的细胞存活很重要，并且它是活性组蛋白甲基转移酶。我们工作的中心假设是SET4是一种应力 - 调节甲基转移酶，该甲基转移酶激活定义的基因表达程序通过其赖氨酸甲基化活性的压力。提出了三个具体目标。在目标一世中，我们会使用生化和质量定义体外和细胞中SET4的底物特异性基于光谱的方法。我们将测试SET4是否主要针对组蛋白，还是还具有额外的非内酮甲基赖氨酸底物，有助于氧化应激回复。在AIM II中，将使用分子和遗传分析来确定途径根据氧化和其他应力调节SET4本身。 set4的机制控制基因表达将通过RNA测序分析和芯片阐明在应力下的一系列突变体中，SET4及其同源甲基的测序。 AIM III会检验以下假设：SET4的PHD手指是SET4 depdent应力反应所必需的促进其定位到染色质。生化和蛋白质组学测定将确定 SET4的PhD手指的组蛋白或非历史结合伴侣，靶向分子和靶向分子和基因组实验将评估PhD手指在定位和活性中的作用染色质的set4。这些研究目的将为A的功能提供大量的见解我们期望的新型表观遗传修饰剂适用于其潜在的人类直系同源物Mll5，与干细胞维持，肿瘤发生和神经发育有关疾病。此外，这项工作将发现环境压力与基于染色质的基因表达调节，这对于我们对我们的理解至关重要异常压力信号传导对基因组的不利影响会导致人类疾病。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Set4 regulates stress response genes and coordinates histone deacetylases within yeast subtelomeres.

DOI：
10.26508/lsa.202101126
发表时间：
2021-12
期刊：
Life science alliance
影响因子：
4.4
作者：
Jethmalani Y;Tran K;Negesse MY;Sun W;Ramos M;Jaiswal D;Jezek M;Amos S;Garcia EJ;Park D;Green EM
通讯作者：
Green EM

Chromatin Immunoprecipitation (ChIP) of Histone Modifications from Saccharomyces cerevisiae.