Regulation of Signaling by Histidine Protein Methylation

组氨酸蛋白甲基化对信号传导的调节

基本信息

批准号：
9974541
负责人：
Or P. Gozani
金额：
$ 31.58万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-15 至 2021-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9974541
关键词：
Actins Arginine Behavior Biochemical Biological Biological Assay Biological Process Biology Biophysics Birth Cardiac Cell Line Cell physiology Cells Cellular biology Chemicals Chemistry Chromatin Cysteine Cytoplasmic Protein Data Disease Ensure Enterovirus Enzymes Event Family Family member Filament Foundations Generations Genetic Glutamine Goals Histidine Human Human Genome Impairment In Vitro Infection Integration Host Factors Knowledge Lysine Malignant Neoplasms Mammalian Cell Mammals Mediating Messenger RNA Methylation Methyltransferase Modeling Modification Molecular Mus Muscle Muscle Contraction Muscle Development Muscle function Mutation Myopathy Myosin ATPase Pathogenesis Pathologic Pathologic Processes Pathology Pathway interactions Phenotype Physiological Post-Translational Protein Processing Process Protein Isoforms Protein Methylation Proteins Proteome Proteomics RNA Splicing Regulation Role SET Domain Signal Transduction Skeletal Muscle Smooth Muscle Smooth Muscle Myocytes Source System Testing Therapeutic Intervention Work behavior in vitro cancer cell cell type chemical reaction experimental study human disease in vivo insight malignant muscle neoplasm muscle physiology new therapeutic target novel polymerization single molecule tool virtual

项目摘要

Abstract Covalent post-translational protein modifications (PTMs) contribute to all aspects of cell physiology and are a primary source of protein functional diversity in mammalian cells. Our overarching goal is to better understand the role of protein methylation signaling in the regulation of diverse biological functions and how disruption in these mechanisms contributes to cancer and other disease pathologies. While the most commonly studied methylation events occur on lysine and arginine residues – glutamine, cysteine, and histidine residues are also methylated – though these modification events are thought to occur on only a limited number of proteins and relatively little is known about the enzymes that catalyze these chemical reactions. In particular, while modification of histidine residues by methylation is thought to be a rare event on proteins, the methylation of actin at histidine 73 (actin-H73me) is a canonical modification in mammals that was identified more than fifty years ago. However, the function of actin-H73me is enigmatic and the enzyme/s generating this abundant modification event are not known. In preliminary work we have identified SETD3 as the first known metazoan protein histidine methyltransferase (PHMT). SETD3, a little studied cytoplasmic protein that belongs to the SET domain family of enzymes, is implicated in processes involved in muscle function and cancer. However, a clear function for SETD3 is not known. Our initial data identified SETD3 as a critical host factor required for infection by a broad class of enteroviruses. We also have evidence for SETD3 regulation of smooth muscle physiology in cells and in vivo. In humans, beyond actin, virtually nothing is known about the molecular, signaling and biological consequences associated with histidine methylation. Our central hypothesis is that histidine methylation of actin and other proteins by enzymes such as SETD3 have an underappreciated and significant role in signal transduction, cell biology, and disease pathogenesis. We propose to use biochemical, cellular, genetic, and proteomic approaches to elucidate the molecular, biological and pathological functions of histidine methylation, with a focus on the biology surrounding SETD3 and SETD3-catalyzed modification of actin. In Aim 1 we will perform experiments to gain a molecular level understanding of the consequence of actin H73 methylation by SETD3. The goal of Aim 2 is to investigate SETD3 cellular functions. Experiments are planned to identify the role of SETD3’s enzymatic activity in actin- related cellular functions and cancer cell phenotypes. We will also explore the molecular mechanisms by which actin is paired with SETD3 for methylation. The goal of Aim 3 is to expand our knowledge of histidine methylation signaling in humans beyond that of SETD3 and actin. We will use computational, proteomic, and biochemical strategies to identify and validate novel human histidine methylated proteins and to discover new histidine methyltransferases.

抽象的共价后翻译蛋白修饰（PTM）有助于细胞生理的各个方面，是一种哺乳动物细胞中蛋白质功能多样性的主要来源。我们的总体目标是更好地了解蛋白甲基化信号传导在调节潜水生物学功能的作用以及如何破坏这些机制有助于癌症和其他疾病病理。虽然最常见的研究甲基化事件发生在赖氨酸和精氨酸残留物上 - 谷氨酰胺，半胱氨酸和组氨酸残留物也是甲基化 - 尽管这些修改事件被认为仅发生在有限数量的蛋白质和关于催化这些化学反应的酶，相关的知识知之甚少。特别是通过甲基化修饰组氨酸残留物被认为是蛋白质的罕见事件，是甲基化的甲基化组氨酸73（肌动蛋白-H73Me）的肌动蛋白是哺乳动物的典型修饰，已鉴定出五十多个几年前。但是，Actin-H73Me的功能是神秘的，并且酶/s生成了此绝对修改事件尚不清楚。在初步工作中，我们确定setd3是第一个已知的后生动物蛋白质组氨酸甲基转移酶（PHMT）。 setd3，属于该集合的小二二拟合细胞质蛋白酶的域家族在参与肌肉功能和癌症的过程中实施。但是，很清楚 setD3的功能尚不清楚。我们的初始数据将SETD3确定为感染所需的关键宿主因素通过一类广泛的肠病毒。我们也有证据表明平滑肌生理的SETD3调节在细胞和体内。在人类中，除了肌动蛋白之外，几乎没有关于分子，信号和与组氨酸甲基化相关的生物学后果。我们的中心假设是组氨酸诸如SetD3等酶对肌动蛋白和其他蛋白质的甲基化不足和显着在信号转移，细胞生物学和疾病发病机理中的作用。我们建议使用生化，细胞，遗传和蛋白质组学方法来阐明分子，组氨酸甲基化的生物学和病理功能，重点是围绕SETD3的生物学和setD3催化的肌动蛋白的修饰。在AIM 1中，我们将执行实验以获得分子水平通过setD3对肌动蛋白H73甲基化的后果的理解。目标2的目的是调查 setD3细胞函数。计划实验以确定setD3酶活性在肌动蛋白中的作用相关的细胞功能和癌细胞表型。我们还将探索分子机制肌动蛋白与setD3配对用于甲基化。目标3的目标是扩大我们对组氨酸的了解人类中的甲基化信号传导超出了setd3和肌动蛋白。我们将使用计算，蛋白质组学和生化策略以识别和验证新型的人组氨酸甲基化蛋白并发现新的组氨酸甲基转移酶。