Structural Studies of SUMO Protein Modification

SUMO蛋白修饰的结构研究

• 批准号: 8266481
• 负责人: CHRISTOPHER D. LIMA
• 金额: $ 39.69万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8266481
• 关键词: Active Sites; Address; Binding; Binding Proteins; Biochemical; Biochemical Genetics; C-terminal; Cell Cycle Progression; Cell Cycle Regulation; Cell physiology; Cells; Chemicals; Chromosome Segregation; Complex; Cysteine; Cytokinesis; DNA Damage; DNA Repair; DNA Replication Factor; DNA biosynthesis; Data; Enzymes; Evolution; Family; Glycine; Health; Human; Inflammation; Ligation; Lysine; M cell; Malignant Neoplasms; Mediating; Metabolism; Mission; Mitosis; Modification; Molecular Conformation; Molecular Weight; Neurodegenerative Disorders; Nuclear; Pathway interactions; Peptide Hydrolases; Phosphorylation; Play; Positioning Attribute; Post-Translational Protein Processing; Process; Property; Protein Isoforms; Proteins; Recruitment Activity; Regulation; Research; Research Proposals; Role; Side; Signal Transduction; Signal Transduction Pathway; Site; Small Ubiquitin-Related Modifier Proteins; Staging; Stress; Structure; Substrate Specificity; Surface; Transcriptional Regulation; Ubiquitin; Ubiquitin Like Proteins; Ubiquitin-Activating Enzymes; Ubiquitination; United States National Institutes of Health; Yeasts; adduct; adenylate; amino group; analog; base; enzyme mechanism; helicase; human disease; member; nucleocytoplasmic transport; protein degradation; protein distribution; protein function; public health relevance; reconstitution; response; structural biology; thioester; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): Signal transduction pathways rely on reversible chemical modifications to relay information within and across cells. Covalent modification of protein substrates by the ubiquitin-like protein SUMO (small ubiquitin-like modifier) contributes to pathways that regulate core cellular functions including nuclear transport, cytokinesis, chromosome segregation, G2-M cell cycle progression and transcriptional regulation among many others. Post-translational modification by ubiquitin (Ub) and ubiquitin-like (Ubl) proteins such as SUMO requires the sequential action of E1 activating enzymes, E2 conjugating enzymes and E3 ligases while Ub/Ubl processing and deconjugation is catalyzed by Ub/Ubl-specific proteases. Because SUMO conjugation plays an integral role in eukaryotic nuclear metabolism and cell cycle control, our studies are of direct relevance to human health, cancer, and the mission of the NIH. This proposal seeks to address the functional significance for components of the SUMO conjugation pathway through structural, biochemical and genetic studies that will establish the basis for 1) SUMO and ubiquitin activation, 2) SUMO conjugation by E2 and E3 enzymes, 3) regulation of SUMO pathway through characterization of SUMO-binding domains and through studies that will determine the importance of SUMO surfaces in response to environmental stress such as DNA damage, 4) address the structural biology associated with SUMO modified PCNA and its recognition by the anti- recombinogenic helicase Srs2. Because the enzymes, mechanisms and co-factors that constitute the SUMO conjugation pathway are conserved or conceptually analogous to those in other Ub/Ubl pathways, our studies will be broadly relevant and will impact research on protein conjugation by Ub and other Ubl modifiers. PUBLIC HEALTH RELEVANCE: Reversible post-translational modifications relay information within and across cells in signal transduction pathways that control the spatial and temporal distribution of protein substrates. Covalent modification of proteins by ubiquitin (Ub) and ubiquitin-like (Ubl) proteins such as SUMO (small ubiquitin-like modifier) and Nedd8 impact nearly all facets of cellular metabolism including cell cycle control, protein degradation, protein localization, nuclear transport, cytokinesis, chromosome segregation, and transcriptional regulation among many others. This proposal seeks to explore the mechanisms that underlie protein modification by SUMO through structural, biochemical and genetic studies that will address 1) Ub/Ubl activation by the E1 enzyme, 2) SUMO conjugation by E2 and E3 enzymes, 3) structural determinants required for recognition of SUMO by SUMO binding proteins, and 4) structural analysis of PCNA, its modification by SUMO, and its recognition by Srs2. The SUMO pathway regulates many pathways that are associated with human disease conditions including neurodegenerative disorders, cancer, and inflammation.

描述（由申请人提供）：信号转导途径依赖于可逆的化学修饰，在细胞内和细胞间传递信息。泛素样蛋白SUMO（小泛素样修饰物）对蛋白质底物的共价修饰有助于调节核心细胞功能的途径，包括核转运、胞质分裂、染色体分离、G2-M细胞周期进展和转录调节等。泛素（Ub）和泛素样（Ubl）蛋白如SUMO的翻译后修饰需要E1激活酶、E2缀合酶和E3连接酶的顺序作用，而Ub/Ubl加工和解缀合由Ub/Ubl特异性蛋白酶催化。由于SUMO结合在真核细胞核代谢和细胞周期控制中起着不可或缺的作用，我们的研究与人类健康、癌症和NIH的使命直接相关。该提案旨在通过结构、生物化学和遗传学研究来解决SUMO缀合途径组分的功能意义，这些研究将为1）SUMO和泛素活化，2）E2和E3酶的SUMO缀合，3）通过表征SUMO-1来调节SUMO途径。结合结构域，并通过研究，将确定SUMO表面的重要性，在响应环境压力，如DNA损伤，4）解决与SUMO修饰的PCNA相关的结构生物学及其被抗重组解旋酶Srs 2识别。由于构成SUMO缀合途径的酶、机制和辅因子与其他Ub/Ubl途径中的酶、机制和辅因子是保守的或在概念上类似，因此我们的研究将具有广泛的相关性，并将影响Ub和其他Ubl修饰剂对蛋白质缀合的研究。 公共卫生关系：可逆的翻译后修饰在信号转导通路中在细胞内和细胞间传递信息，控制蛋白质底物的空间和时间分布。泛素（Ub）和泛素样（Ubl）蛋白如SUMO（小泛素样修饰物）和Nedd 8对蛋白质的共价修饰几乎影响细胞代谢的所有方面，包括细胞周期控制、蛋白质降解、蛋白质定位、核转运、胞质分裂、染色体分离和转录调控等。本提案旨在通过结构、生物化学和遗传学研究探索SUMO修饰蛋白质的机制，这些研究将解决1）E1酶激活Ub/Ubl，2）E2和E3酶的SUMO结合，3）SUMO结合蛋白识别SUMO所需的结构决定因素，以及4）PCNA的结构分析，SUMO修饰及其Srs 2识别。SUMO通路调节许多与人类疾病相关的通路，包括神经退行性疾病、癌症和炎症。