Function of SMYD lysine methyltransferases in stress responses and proteostasis

SMYD 赖氨酸甲基转移酶在应激反应和蛋白质稳态中的功能

基本信息

批准号：
10745041
负责人：
Erin Green
金额：
$ 31.02万
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：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10745041
关键词：
Address Age of Onset Aging Biochemical Biological Assay Biological Models Biological Process Blood Vessels Cell Aging Cell physiology Cells Chemicals Code Data Degradation Pathway Deterioration Disease Enzymes Etiology Family Genetic Genetic Models Goals Heart Diseases Histones Human In Vitro Investigation Knowledge Link Lysine Methylation Methyltransferase Modeling Modification Molecular Molecular Chaperones Muscle Muscle Development Muscular Atrophy Myocardium Nerve Degeneration Neurons Orphan Outcome Pathology Pathway interactions Physiological Post-Translational Protein Processing Protein Biosynthesis Proteins Proteome Proteomics Publishing Quality Control Regulation Role Saccharomycetales Series Skeletal Muscle Stress Structure Substrate Specificity Testing Translations Work Yeasts age related biological adaptation to stress cell age experience experimental study functional outcomes genetic approach in vitro activity insight non-histone protein novel overexpression prevent protein TDP-43 protein aggregation protein function protein protein interaction proteostasis senescence targeted treatment tool

项目摘要

PROJECT SUMMARY The deterioration of protein homeostasis is a signature of aging cells and underlies the etiology of numerous aging-related diseases. Proteostasis is normally maintained through a well-coordinated network of factors that include protein synthesis regulators, molecular chaperones that promote proper folding, quality control and proteolytic degradation machinery. This proteostasis network serves to prevent the accumulation of misfolded, non-functional, and aggregated proteins, which is particularly critical during stress and as cells age. Proteins which function within the proteostasis network are highly-regulated by different mechanisms including their post- translational modification. The modification lysine methylation has been identified on many factors that contribute to proteostasis, though the molecular role for methylation within the proteostasis network remains poorly-defined. The conserved SMYD family of lysine methyltransferases are known to methylate factors important to proteome integrity, such as chaperones, although their function in maintaining proteostasis is not well-understood. We have investigated the function of an orphan yeast SMYD lysine methyltransferase, Set6, and determined that it is a critical regulator of the proteostasis network via its methyltransferase activity. The overarching goal of the proposed work is to identify the specific regulatory role for Set6 in proteostasis and to define its catalytic activity on factors important for proteome integrity, including the cytosolic chaperone Hsp70. To address this, we propose to use a series of molecular and cell biological assays to dissect the molecular contribution of Set6 and its catalytic activity to the proteostasis network under normal and stress conditions. We will also use quantitative proteomics, structure-function analysis, and genetic approaches to define regulatory mechanisms directing the activity of Set6 under normal and stress conditions, with a focus on its protein-protein interactions. Finally, we will define the molecular consequences of Set6-dependent methylation on Hsp70 using molecular, biochemical, and genetic approaches, and test the hypothesis that methylation by Set6 alters the Hsp70 interactome during stress. We will also investigate whether Set6 has additional substrates in the proteostasis network using biochemical, proteomic, and genetic assays. Altogether, these studies will reveal new regulatory mechanisms governing the proteostasis network and will advance our understanding of the roles for SMYD enzymes in proteome integrity. This work will also provide insight into aging-related pathologies characterized by misfolded or aggregated proteins and may uncover new mechanisms that can be targeted therapeutically to promote healthy proteostasis as cells age and prevent age-dependent diseases.

项目摘要蛋白质稳态的恶化是衰老细胞的特征，是许多众多病因的基础与衰老有关的疾病。通常通过良好协调的因素网络来维护蛋白质静脉曲张包括蛋白质合成调节剂，促进适当折叠的分子伴侣，质量控制和蛋白水解降解机械。该蛋白质的网络有助于防止错误折叠的积累，非功能性和聚集的蛋白质，这在压力和细胞年龄时特别关键。蛋白质在蛋白质之间的网络中哪些功能受到不同机制的高度调节翻译修改。修饰赖氨酸甲基化已在许多有助于的因素上鉴定对于蛋白质，尽管蛋白抑制网络内甲基化的分子作用仍然很差。众所周知，赖氨酸甲基转移酶的保守SMYD家族对蛋白质组很重要尽管伴侣在维持蛋白质的功能中的功能并不是很好的理解。我们已经研究了孤儿酵母Smyd赖氨酸甲基转移酶Set6的功能，并确定了它是通过其甲基转移酶活性的蛋白抑制网络的关键调节剂。总体目标拟议的工作是确定SET6在蛋白质的特定调节作用，并定义其催化活性关于蛋白质组完整性重要的因素，包括胞质伴侣HSP70。为了解决这个问题，我们建议使用一系列分子和细胞生物学测定法剖定SET6和在正常和应力条件下，其催化活性归因于蛋白质的网络。我们还将使用定量蛋白质组学，结构功能分析和遗传方法定义指导的调节机制 SET6在正常和应力条件下的活性，重点是其蛋白质 - 蛋白质相互作用。最后，我们将使用分子，生化，依赖set6依赖性甲基化在HSP70上的分子后果和遗传方法，并检验以下假设：set6甲基化改变了HSP70在压力。我们还将调查SET6使用proteeostasis网络中是否具有其他底物生化，蛋白质组学和遗传测定法。总之，这些研究将揭示新的调节机制管理Proteostasis网络，并将促进我们对Smyd酶在中的作用的理解蛋白质组完整性。这项工作还将提供有关衰老相关的病理的见解或聚集的蛋白质，可能会发现可以针对治疗的新机制以促进健康的蛋白质抑制作用是细胞的年龄并预防年龄依赖性疾病。