The SMYD lysine methyltransferase Set6 in signaling and proteostasis

SMYD 赖氨酸甲基转移酶 Set6 在信号传导和蛋白质稳态中的作用

• 批准号: 9808970
• 负责人: Erin Green
• 金额: $ 21.93万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE COUNTY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-15 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9808970
• 关键词: Actins; Address; Aging; Biochemical; Biochemical Genetics; Biological Assay; Biological Models; Biological Process; Cell Aging; Cell Survival; Cells; Client; Complex; Data; Deterioration; Disease; Enzymes; Equilibrium; Event; Family; Genetic; Genetic Models; Heart Diseases; Histone H4; Histones; Human; Investigation; Knowledge; Light; Link; Lysine; Maintenance; Mass Spectrum Analysis; Mediating; Methylation; Methyltransferase; Molecular; Molecular Chaperones; Molecular Genetics; Muscle Development; Muscular Atrophy; Myocardium; Pathology; Pathway interactions; Positioning Attribute; Post-Translational Protein Processing; Process; Protein Biosynthesis; Protein Methylation; Proteins; Proteome; Proteomics; Regulation; Reporting; Resolution; Role; SET Domain; SMYD3 gene; Saccharomyces; Saccharomyces cerevisiae; Saccharomycetales; Signal Transduction; Site; Skeletal Muscle; Stress; Substrate Domain; System; Tertiary Protein Structure; Testing; Therapeutic Intervention; Tubulin; Validation; Work; Yeasts; Zinc Fingers; age related; base; experience; genetic analysis; healthy aging; insight; muscle aging; new therapeutic target; non-histone protein; prefoldin; prevent; protein aggregate; protein aggregation; protein degradation; protein folding; protein function; protein misfolding; protein protein interaction; proteostasis; tool

PROJECT SUMMARY Protein homeostasis, or proteostasis, is achieved through a regulated network of the translational machinery, molecular chaperones, and protein degradation factors, which function to maintain the abundance of the cell’s proteins in folded, soluble, non-aggregated states. Deterioration of the proteostasis network is a hallmark of aging cells and underlies numerous diseases associated with aging. A key mechanism regulating factors within the proteostasis network is their post- translational modification, including methylation at lysine residues. The SMYD family of lysine methyltransferases has been shown to interact with molecular chaperones in diverse systems, and there are numerous reported lysine methylation sites on factors that contribute to proteostasis. However, our knowledge of the methyl-lysine substrates targeted by SMYD methyltransferases and their functional interactions with the proteostasis network is still very limited. The primary objective of the proposed work is to leverage proteomic and molecular tools in the model system Saccharomyces cereviasie to define the substrates for the yeast SMYD enzyme Set6, a largely uncharacterized protein, and determine its molecular function in regulating the proteostasis network. In Aim I, we will use high-resolution mass spectrometry to identify targets of Set6 proteome-wide and perform molecular and biochemical characterization of lysine methylation on the newly-identified substrates. In Aim II, we will use molecular and genetic tools to define the role of Set6 in interacting with and regulating the proteostasis network to prevent the accumulation of unfolded proteins. In total, this work will integrate proteomic and genetic analyses to advance our understanding of the mechanistic links between the SMYD family lysine methyltransferases and molecular chaperones. We expect this work to provide new insights into regulatory mechanisms critical to proteostasis, which may shed light on new targets for therapeutic interventions into diseases associated with protein misfolding and aggregation. Furthermore, these studies will also develop a well-characterized, genetic model system for dissecting the molecular mechanisms by which SMYD enzymes contribute to proteostasis and determining how their manipulation may prevent aging-associated pathologies.

项目概要 蛋白质稳态或蛋白质稳态是通过翻译调节网络实现的 机械、分子伴侣和蛋白质降解因子，其功能是维持 细胞蛋白质的丰度处于折叠、可溶、非聚集状态。恶化 蛋白质稳态网络是衰老细胞的标志，也是许多相关疾病的基础 随着衰老。蛋白质稳态网络内调节因素的一个关键机制是它们的后 翻译修饰，包括赖氨酸残基的甲基化。赖氨酸的 SMYD 家族 甲基转移酶已被证明与不同系统中的分子伴侣相互作用， 并且有许多报道的赖氨酸甲基化位点影响因素 蛋白质稳态。然而，我们对 SMYD 靶向的甲基赖氨酸底物的了解 甲基转移酶及其与蛋白质稳态网络的功能相互作用仍然非常重要 有限的。拟议工作的主要目标是利用蛋白质组学和分子工具 在酿酒酵母模型系统中定义酵母 SMYD 的底物 酶 Set6，一种很大程度上未表征的蛋白质，并确定其在调节中的分子功能 蛋白质稳态网络。在目标 I 中，我们将使用高分辨率质谱法来识别 Set6 蛋白质组范围内的靶标，并对赖氨酸进行分子和生化表征 新鉴定的底物上的甲基化。在目标 II 中，我们将使用分子和遗传工具 定义 Set6 在与蛋白质稳态网络相互作用和调节中的作用，以防止 未折叠蛋白质的积累。总的来说，这项工作将整合蛋白质组学和遗传分析 促进我们对 SMYD 家族赖氨酸之间机制联系的理解 甲基转移酶和分子伴侣。我们期望这项工作能够提供新的见解 对蛋白质稳态至关重要的调控机制，这可能有助于揭示蛋白质稳态的新目标 对与蛋白质错误折叠和聚集相关的疾病的治疗干预。 此外，这些研究还将开发一个特征良好的遗传模型系统 剖析 SMYD 酶促进蛋白质稳态的分子机制 确定它们的操作如何预防与衰老相关的病变。