SUMO-dependent Regulation of Ubiquitin Ligases in Genomic Stability

基因组稳定性中泛素连接酶的 SUMO 依赖性调节

• 批准号: 8206797
• 负责人: MICHAEL N BODDY
• 金额: $ 37.22万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8206797
• 关键词: Acute Promyelocytic Leukemia; Address; Apoptosis; Apoptotic; Arsenic Trioxide; Biochemical; Biochemical Genetics; Biochemistry; Biological; Biology; Camptothecin; Cell Line; Cells; Cellular biology; Complex; Coupled; Crystallography; DNA Repair; DNA Repair Pathway; DNA repair protein; Data; Enzymes; Family; Family member; Fission Yeast; Foundations; Functional disorder; Future; Gatekeeping; Genetic; Genome; Genome Stability; Genomic Instability; Genomics; Genotoxic Stress; Human; Hyperactive behavior; In Vitro; Knowledge; Letters; Link; Maintenance; Malignant Neoplasms; Mammalian Cell; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Molecular; Mus; Nuclear; Pathway interactions; Patients; Poison; Predisposition; Protein Family; Proteins; Proteome; Proteomics; Recruitment Activity; Recycling; Regulation; Resistance; Resolution; Risk; Role; Solutions; Stimulus; Structure; Sumoylation Pathway; Testing; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Topoisomerase; Toxic Environmental Substances; Tumor Suppressor Proteins; Type I DNA Topoisomerases; Ubiquitin; Ubiquitination; Yeasts; base; cancer genomics; carcinogenesis; casein kinase II; cell growth; environmental mutagens; human disease; in vivo; insight; multicatalytic endopeptidase complex; mutant; novel; protein function; research study; response; tumorigenesis; ubiquitin ligase; ubiquitin-protein ligase; yeast genetics

Ubiquitin and the small ubiquitin-like modifier (SUMO), have critical but classically distinct roles in genome stability and responses to environmental toxins. Ubiquitin and SUMO are covalently attached by E3 ligases to target proteins, to regulate the cell's proteome dynamically. Ubiquitin and SUMO pathway dysfunction results in genetic instability and cancer predispositions in humans. We recently discovered the SUMO-targeted E3 ubiquitin ligase family (STUbLs), which ubiquitinate and target sumoylated proteins to the proteasome for degradation. This unanticipated phenomenon requires a revision to the spectrum of SUMO functions. This revised proposal builds upon our discovery and adds new data that directly connects the STUbL mechanism to cancer and genomic stability. We have identified the main mediators of genomic instability in fission yeast STUbL mutants and we will characterize the cellular and molecular mechanisms of STUbLs through these targets. Results from fission yeast will provide a framework for studies conducted on the human and mouse STUbL targets. Notably, these targets place the STUbL mechanism of action at the intersection of cell growth/apoptotic pathways, genomic stability and oncogenesis. Our two Specific Aims focus on defining the molecular mechanisms of STUbL function both in vitro and in vivo. In Aim 1, we will test the hypothesis that STUbLs act as gatekeepers for apoptosis versus DNA repair pathways, by controlling the fates of key SUMO conjugates. In Aim 2, we will address our overall hypothesis that STUbLs are cellular master keys, controlling genome stability through specific and definable protein interaction interfaces. To attain our Aims we will integrate powerful yeast genetics with cutting-edge structural analyses, MudPIT mass spectrometry, biochemistry and mammalian cell biology. The expected results will provide critical mechanistic insights into the genome stabilizing roles of STUbLs including RNF4, and may suggest therapeutic strategies for RNF4- linked human diseases. Additionally, a detailed understanding of STUbL function should yield general implications for responses to genotoxic stress.

泛素和小型泛素样修饰剂（SUMO）在基因组中具有关键但经典的作用 稳定性和对环境毒素的反应。泛素和相扑由E3连接酶共价附加到 靶蛋白，以动态调节细胞的蛋白质组。泛素和相扑途径功能障碍结果 在人类的遗传不稳定性和癌症易感性中。我们最近发现了靶向的E3 泛素连接酶家族（坚固），泛素化和靶向sumoy蛋白靶向蛋白酶体 降解。这种意外的现象需要对Sumo功能的光谱进行修订。这 修订的建议基于我们的发现，并添加了直接将坚定机制连接到的新数据 癌症和基因组稳定性。我们已经确定了裂变酵母中基因组不稳定性的主要介体 坚定的突变体，我们将通过这些表征稳固的细胞和分子机制 目标。裂变酵母的结果将为对人和小鼠进行的研究提供一个框架 坚定的目标。值得注意的是，这些目标将作用机理置于细胞的交集 生长/凋亡途径，基因组稳定性和肿瘤发生。我们的两个具体目标着重于定义 体外和体内的强大功能的分子机制。在AIM 1中，我们将检验以下假设 坚定用作凋亡与DNA修复途径的守门人，通过控制钥匙相扑的命运 共轭。在AIM 2中，我们将解决我们的总体假设，即坚定是蜂窝主键，控制 通过特定且可定义的蛋白质相互作用界面的基因组稳定性。为了实现我们的目标，我们将 将强大的酵母遗传学与最先进的结构分析，泥浆质谱法相结合， 生物化学和哺乳动物细胞生物学。预期的结果将为关键的机械见解 包括RNF4在内的坚定作用的基因组稳定作用，并可能提出RNF4-的治疗策略 连接的人类疾病。此外，对持久功能的详细理解应产生一般性 对遗传毒性应激反应的影响。