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）：项目概述SUMO（小泛素相关修饰物）是与其他蛋白质后结合的蛋白质，从而调节对正常细胞生长、分化和存活至关重要的广泛的基本功能。值得注意的是，类小泛素化与多种人类疾病的发展和进展直接相关，包括神经变性、中风、心脏病、唇腭裂和癌症。在过去的十年中，在阐明SUMO与靶蛋白结合的分子机制和鉴定受SUMO化调控的细胞过程方面取得了巨大进展。然而，尽管有这些见解，关于类小泛素化如何在分子水平上影响修饰的蛋白质，以及这些影响如何转化为细胞功能的控制等基本问题仍有待解决。在许多情况下，类小泛素化对靶蛋白的影响仍然未知。此外，SUMO化如何能够在与不同蛋白质缀合后引发独特的命运也知之甚少。我们将使用芽殖酵母，S。酿酒酵母，作为一种模式生物研究的基本功能sumoylation。提出的研究的一个目标是开发一个高分辨率的SUMO结构/功能图。这一目标将通过酵母SUMO突变体的综合库的合成和这些突变体的表征，使用各种检测设计的探针SUMO结构/功能的关系，生物化学和遗传。有了这个库，我们将测试的假设，SUMO代表一个通用的和可变的信号与不同的表面残基影响与不同的调控因子和下游SUMO结合蛋白的相互作用。我们还将测试的假设，sumoylation功能的伴侣样能力，以保护蛋白质的损伤和聚集所造成的细胞毒性细胞应激的影响。