High throughput structure/function analysis of SUMO modification

SUMO修饰的高通量结构/功能分析

• 批准号: 8727047
• 负责人: MICHAEL J. MATUNIS
• 金额: $ 30.78万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8727047
• 关键词: Address; Affect; Alanine; Alleles; Amino Acid Substitution; Amino Acids; Animal Model; Binding; Binding Proteins; Biochemical; Biological Assay; Cell Differentiation process; Cell physiology; Cellular Stress; Charge; Chromosome Segregation; Collection; DNA Repair; Databases; Defect; Development; Disease; Evaluation; Genes; Genetic; Genetic Transcription; Goals; Growth; Heart Diseases; Hypoxia; Individual; Libraries; Link; Malignant Neoplasms; Malignant Palate Neoplasm; Maps; Measures; Microarray Analysis; Mitosis; Modification; Molecular; Molecular Analysis; Molecular Chaperones; Mutation; Natural Killer Cells; Nerve Degeneration; Normal Cell; Oxidative Stress; Pattern; Phenotype; Plasmids; Post-Translational Protein Processing; Process; Production; Property; Proteins; RNA Splicing; Research Design; Resolution; Saccharomyces cerevisiae; Saccharomycetales; Serine; Signal Transduction; Small Ubiquitin-Related Modifier Proteins; Stroke; Structure; Structure-Activity Relationship; Surface; Synthetic Genes; Techniques; Testing; Translating; Ubiquitin; Yeasts; biophysical properties; cell growth; cleft lip and palate; design; gene interaction; genetic analysis; high throughput analysis; human disease; in vitro Assay; in vivo; insight; isopeptidase; mRNA Precursor; mutant; protein misfolding; response; synthetic construct; temperature sensitive mutant

DESCRIPTION (provided by applicant): PROJECT SUMMARY SUMOs (small ubiquitin-related modifiers) are proteins that are posttranslationally conjugated to other proteins, thereby regulating a wide range of essential functions critical for normal cell growth, differentiation and survival. Notably, sumoylation has been directly linked to the development and progression of a variety of human diseases, including neurodegeneration, stroke, heart disease, cleft lip/palate and cancer. During the past ten years enormous progress has been made in elucidating the molecular mechanisms involved in conjugating SUMOs to target proteins and in identifying the cellular processes regulated by sumoylation. Despite these insights, however, fundamental questions about how sumoylation actually affects modified proteins at the molecular level, and how these effects are translated into control of cell function, remain to be addressed. In many cases, the effects of sumoylation on target proteins remain unknown. In addition, how sumoylation is able to elicit unique fates upon conjugation to different proteins is also poorly understood. We will use the budding yeast, S. cerevisiae, as a model organism for studying the essential functions of sumoylation. A goal of the proposed studies is develop a high-resolution structure/function map of SUMO. This goal will be achieved through the synthesis of a comprehensive library of yeast SUMO mutants and characterization of these mutants using a variety of assays designed to probe SUMO structure/function relationships both biochemically and genetically. With this library, we will test the hypothesis that SUMO represents a versatile and variable signal with distinct surface residues affecting interactions with distinct regulatory factors and downstream SUMO-binding proteins. We will also test the hypothesis that sumoylation functions in a chaperone-like capacity to protect proteins from damage and aggregation caused by the effects of cytotoxic cell stresses.

描述（由申请人提供）：项目摘要 SUMO（小泛素相关修饰剂）是翻译后与其他蛋白质缀合的蛋白质，从而调节对正常细胞生长、分化和存活至关重要的广泛基本功能。值得注意的是，SUMO化与多种人类疾病的发生和进展直接相关，包括神经变性、中风、心脏病、唇裂/腭裂和癌症。在过去的十年中，在阐明 SUMO 与靶蛋白结合的分子机制以及识别 SUMO 化调节的细胞过程方面取得了巨大进展。然而，尽管有这些见解，关于苏酰化实际上如何在分子水平上影响修饰蛋白以及这些影响如何转化为对细胞功能的控制的基本问题仍有待解决。在许多情况下，苏酰化对靶蛋白的影响仍然未知。此外，人们对苏酰化如何在与不同蛋白质缀合时引发独特的命运也知之甚少。我们将使用芽殖酵母（酿酒酵母）作为模型生物来研究苏糖化的基本功能。拟议研究的目标是开发 SUMO 的高分辨率结构/功能图。这一目标将通过合成酵母 SUMO 突变体的综合文库并使用各种旨在从生化和遗传学方面探讨 SUMO 结构/功能关系的测定来表征这些突变体来实现。通过这个库，我们将测试以下假设：SUMO 代表一种多功能且可变的信号，具有不同的表面残基，影响与不同调节因子和下游 SUMO 结合蛋白的相互作用。我们还将测试这样的假设：苏酰化以类似伴侣的能力发挥作用，以保护蛋白质免受细胞毒性细胞应激影响引起的损伤和聚集。