Role of Ubiquitin Like Protein ISG15 Conjugation

泛素样蛋白 ISG15 缀合的作用

• 批准号: 7533961
• 负责人: DONG-ER ZHANG
• 金额: $ 32.06万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7533961
• 关键词: Address; Affinity Chromatography; Antibodies; Bacteria; Bacterial Infections; Biological Process; C-terminal; Cell Cycle; Cell physiology; Complex; DNA Repair; Defect; Development; Endocytosis; Enzymes; Funding; Genes; Genetic Transcription; Goals; Housing; Human; Immune; Immune response; Infection; Interferon Type I; Interferons; Knockout Mice; Knowledge; Link; Malignant Neoplasms; Mass Spectrum Analysis; Membrane; Methods; Modification; Molecular; Mus; Nematoda; Organism; Pathology; Pathway interactions; Peptides; Play; Post-Translational Protein Processing; Prevention; Proteins; Quality Control; Receptor Signaling; Role; Signal Pathway; Signal Transduction; Site; Stress; Testing; Therapeutic; Ubiquitin Like Proteins; Ubiquitination; Viral; Virus; Virus Diseases; Yeasts; high throughput screening; human disease; insight; pathogen; protein function; response

The long-term goal of this study is to understand the biological function of protein modification by the ubiquitin like protein ISG15. In recent years our knowledge of protein ubiquitination (ubiquitylation) has expanded rapidly and as a consequence protein ubiquitylation has been found to play important roles in various aspects of cellular function, including the cell cycle, membrane receptor signal transduction, endocytosis, protein quality control, transcription, and DNA repair. In contrast, little is known about the role of protein modification by ISG15. ISG15 is encoded by an interferon stimulated gene. Its expression is highly upregulated by Type I interferon and by bacterial and viral infections. Since ISG15 is not found in simple eukaryotic organisms, such as yeast and nematodes, it is unlikely to be a house keeping gene. Instead, it should be involved in specialized functions in complex organisms, such as human and mouse. In the past four year funding period, we have identified a group of ISG15 targets, characterized the effect of ISG15 modification on some of these targets, identified the ISG15 E2 and a few of the E3 enzymes, and established ISG15 E1 knockout mice. This proposal for the next two-year funding period will test the hypothesis that protein ISG15 modification plays an important role in modulating cellular function during immune responses. The studies proposed in Specific Aim #1 will identify ISG15 modification sites via high throughput screening of ISGylated proteins. The studies proposed in Specific Aim #3 will characterize ISG15 activating enzyme UBE1L knockout mice to examine molecular mechanisms of the function of protein ISGylation. These proposed studies will address important questions about protein ISGylation and may provide valuable insights into the prevention and therapeutic treatment of human diseases, such as pathogen infections and immune defects.

本研究的长期目标是了解泛素样蛋白ISG15修饰蛋白质的生物学功能。近年来，蛋白质泛素化（ubiquitylation，ubiquitylation）的研究进展迅速，其在细胞周期、膜受体信号转导、胞吞作用、蛋白质质量控制、转录和DNA修复等细胞功能中发挥着重要作用。相比之下，关于ISG15对蛋白质修饰的作用知之甚少。ISG15由干扰素刺激基因编码。其表达被I型干扰素以及细菌和病毒感染高度上调。由于ISG15不存在于简单的真核生物中，如酵母和线虫，因此它不太可能是一个管家基因。相反，它应该参与复杂生物体（如人类和小鼠）的专门功能。在过去的四年资助期内，我们已经确定了一组ISG15靶标，表征了ISG15修饰对其中一些靶标的影响，确定了ISG15 E2和一些E3酶，并建立了ISG15 E1敲除小鼠。这项为期两年的资助计划将测试蛋白ISG15修饰在免疫反应中调节细胞功能方面发挥重要作用的假设。具体目标#1中提出的研究将通过高通量筛选ISGylated蛋白来鉴定ISG15修饰位点。具体目标#3中提出的研究将表征ISG15活化酶UBE1L敲除小鼠，以检查蛋白ISG化功能的分子机制。这些拟议的研究将解决有关蛋白质ISGylation的重要问题，并可能为预防和治疗人类疾病（如病原体感染和免疫缺陷）提供有价值的见解。