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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.

项目摘要在波动的环境信号存在下，细胞的存活严重依赖于快速的基因表达的变化。染色质修饰酶是基因组的关键调节因子在应激过程中的重编程，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。等结果有助于病理过程，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起着关键作用，但仍然存在大量的我们对组蛋白修饰的新位点及其对基因组的影响的认识存在空白监管，特别是在环境中遇到的各种压力的存在。我们初步研究发现，酿酒酵母蛋白Set 4，一种潜在的人蛋白MLL 5的直系同源物，对于氧化应激中的细胞存活是重要的，并且它是一种活性组蛋白甲基转移酶我们工作的中心假设是Set 4是一个重音- 受调节的甲基转移酶，其激活确定的基因表达程序，以响应通过其赖氨酸甲基化活性胁迫。提出了三个具体目标。在Aim I中，我们将使用生物化学和质谱技术在体外和细胞中确定Set 4的底物特异性基于光谱法的方法。我们将测试Set 4是否主要靶向组蛋白，或者是否也有额外的非组蛋白甲基赖氨酸底物，反应在目标II中，将使用分子和遗传分析来确定调节Set 4自身对氧化和其他应激的反应。第4组对照基因表达将通过RNA测序分析和chIP- 在胁迫下的一系列突变体中Set 4及其同源甲基标记的测序。Aim III将通过以下方式检验Set 4的PHD指是Set 4依赖性应激反应所需的假设：促进其定位于染色质。生物化学和蛋白质组学分析将确定组4的PHD指的组蛋白或非组蛋白结合配偶体，以及靶向分子和基因组实验将评估PHD指在定位和活性中的作用，在染色质处设置4。这些研究目标将提供实质性的洞察力的功能，我们预期可应用于其潜在的人类直系同源物MLL 5的新型表观遗传修饰剂，它与干细胞维持、肿瘤发生和神经发育有关紊乱此外，这项工作将揭示环境压力与环境压力之间的新联系。基于染色质的基因表达调控，这将是至关重要的，我们了解异常压力信号对基因组的错误调节如何导致人类疾病。

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.

SET4调节应力反应基因，并在酵母亚端子中协调组蛋白脱乙酰基酶。

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

Set4 is a chromatin-associated protein, promotes survival during oxidative stress, and regulates stress response genes in yeast.

Set4 是一种染色质相关蛋白，可促进氧化应激期间的存活，并调节酵母中的应激反应基因。