High throughput structure/function analysis of SUMO modification

SUMO修饰的高通量结构/功能分析

• 批准号: 8328625
• 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/8328625
• 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; 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Sumos（与泛素相关的小修饰剂）是诱导后与其他蛋白质共轭的蛋白质，从而调节对正常细胞生长，分化和生存至关重要的广泛基本功能。值得注意的是，Sumoylation已直接与各种人类疾病的发展和发展有关，包括神经退行性，中风，心脏病，唇裂/pa/paper和癌症。在过去的十年中，在阐明与sumos相结合的分子机制方面取得了巨大进展，并在识别由sumoylation调节的细胞过程中。但是，尽管有这些见解，但有关Sumoylation如何实际影响分子水平上的修饰蛋白的基本问题，以及如何将这些效应转化为细胞功能的控制，仍然有待解决。在许多情况下，Sumoylation对靶蛋白的影响仍然未知。此外，Sumoylation如何在与不同蛋白质结合时能够引起独特的命运。我们将使用发芽的酵母菌酿酒酵母作为模型生物，用于研究sumoylation的基本功能。提出的研究的目标是开发SUMO的高分辨率结构/功能图。通过综合酵母相扑突变体的综合文库以及这些突变体的表征，使用旨在探测Sumo结构/功能关系的各种测定法，从生化和遗传上探测这些突变体的表征。使用此库，我们将测试以下假设：Sumo代表一种多功能和可变的信号，其表面残基有不同的表面残基，影响与不同的调节因子和下游相互结合蛋白的相互作用。我们还将检验以下假设：Sumoylation在伴侣样能力下起作用，以保护蛋白质免受细胞毒性细胞应激的影响引起的损伤和聚集。