Regulation and Function of SUMO Protein Modification -Equipment Supplement

SUMO蛋白修饰的调节与作用-设备补充

• 批准号: 10581055
• 负责人: MICHAEL J. MATUNIS
• 金额: $ 22.97万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10581055
• 关键词: Address; Cell Line; Cell physiology; Cells; Cellular Morphology; Cellular Stress; Defect; Disease; Equipment; Gene Expression; Health; Human; Hypersensitivity; Knock-out; Knowledge; Link; Malignant Neoplasms; Molecular; Molecular Mechanisms of Action; Neurodegenerative Disorders; Pathway interactions; Phase I Clinical Trials; Phenotype; Post-Translational Protein Processing; Property; Proteins; Regulation; Role; Signal Transduction; Small Ubiquitin-Related Modifier Proteins; Sumoylation Pathway; Testing; Therapeutic; Ubiquitin; Vertebrates; cancer therapy; human disease; inhibitor; insight; interest; mutant; paralogous gene; proteostasis

PROJECT SUMMARY Small ubiquitin related modifiers (SUMOs) function as posttranslational protein modifications and thereby regulate nearly all essential cell functions. As such, sumoylation is linked to a variety of human diseases, including cancer and neurodegenerative disorders, and there is great interest in targeting the SUMO pathway for therapeutic purposes. Notably, the first phase I clinical trial of a SUMO inhibitor was recently approved for cancer therapy. A more detailed understanding of basic molecular mechanisms regulating sumoylation and its consequences on cell function, however, is needed to achieve the full potential of these efforts. In particular, an ability to target specific branches of the SUMO pathway could allow for more precise therapies. Vertebrates express multiple SUMO paralogs which could allow for such precision targeting. However, the unique properties and functions of SUMO paralogs remain poorly understood. To address this gap in knowledge, we are taking advantage of recently developed SUMO1 and SUMO2 knockout cell lines to define and characterize the specific functions and molecular mechanisms of action of these two paralogs. Aims of our proposal include: (1) We will obtain support for the hypothesis that SUMO1 and SUMO2 function as unique signals with non- redundant roles in regulating distinct cellular processes. This will be achieved through characterization of paralog-specific phenotypes we have identified in SUMO1 and SUMO2 knockout cell lines, including changes in gene expression, cell morphology, defects in proteostasis and hypersensitivities to cell stress. (2) We will test multiple hypotheses to reveal the molecular basis for non-redundant functions of SUMO1 and SUMO2. Hypotheses concerning the function of chain formation and selective non-covalent interactions with effector proteins will be explored through rescue of knockout cell phenotypes using a panel of SUMO1 and SUMO2 mutant proteins. (3) We will test the hypothesis that phenotypic changes observed in SUMO1 and SUMO2 knockout cell lines are due in part to paralog-specific effects on gene expression. Results from our studies will provide a comprehensive understanding of the unique, paralog-specific functions of SUMO1 and SUMO2 and vital insights required to target specific branches of the SUMO pathway for therapeutic purposes.

项目总结 小泛素相关修饰物(SUMO)起翻译后蛋白质修饰的作用，从而 调节几乎所有必要的细胞功能。因此，相思甲基化与多种人类疾病有关， 包括癌症和神经退行性疾病，人们对相扑通路的靶向非常感兴趣 用于治疗目的。值得注意的是，相扑抑制剂的第一阶段临床试验最近被批准用于 癌症治疗。更详细地了解调节相思甲基化的基本分子机制及其 然而，为了充分发挥这些努力的潜力，需要对细胞功能产生影响。特别是，一个 针对相扑路径特定分支的能力可以实现更精确的治疗。脊椎动物 表达多个相扑并行，这可能允许如此精确的目标。然而，独特的 相扑对偶的性质和功能仍然知之甚少。为了解决这一知识差距，我们 正在利用最近开发的SUMO1和SUMO2基因敲除细胞系来定义和表征 这两个类似物的特定功能和分子作用机制。我们建议的目标包括： (1)我们将获得对SUMO1和SUMO2作为唯一信号的假设的支持 在调节不同的细胞过程中扮演多余的角色。这将通过以下方式实现 我们在SUMO1和SUMO2基因敲除细胞系中发现的Paralog特异性表型，包括变化 在基因表达、细胞形态、蛋白平衡缺陷和对细胞应激的敏感性方面。(2)我们会 测试多个假设以揭示SUMO1和SUMO2非冗余功能的分子基础。 关于链形成功能和与效应器的选择性非共价相互作用的假说 将通过使用一组SUMO1和SUMO2抢救敲除细胞表型来探索蛋白质 突变蛋白质。(3)我们将检验在SUMO1和SUMO2中观察到的表型变化的假设 基因敲除细胞系的部分原因是Paralog对基因表达的特异性影响。我们的研究结果将 全面了解SUMO1和SUMO2独特的Paralog特定功能以及 为了治疗的目的，针对相扑通路的特定分支所需的重要见解。