Identifying the targets of oncogenic/tumor-suppressive F box proteins

鉴定致癌/肿瘤抑制 F 盒蛋白的靶标

• 批准号: 9016501
• 负责人: David Paul Toczyski
• 金额: $ 16.19万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9016501
• 关键词: Address; Affect; Affinity; Alleles; Binding; Biology; Cell Cycle; Cell Line; Cells; Complex; Cullin Proteins; DNA Damage; Data; Dissociation; Drug Targeting; F-Box Proteins; FBXW7 gene; Generations; Genes; Genome; Half-Life; Health; Human; Immunoprecipitation; Individual; Ligase; Link; Mass Spectrum Analysis; Mediating; Methods; Multiple Myeloma; Mutagenesis; Mutate; Mutation; Oncogenic; Phosphotransferases; Polyubiquitin; Process; Protein Analysis; Proteins; Regulation; SKP Cullin F-Box Protein Ligases; SKP2 gene; Specificity; Structure; Surface; Technology; Tumor Suppressor Genes; Ubiquitin; Ubiquitination; WD Repeat; Western Blotting; Work; Yeasts; base; cell growth regulation; inhibitor/antagonist; knock-down; member; mutant; novel; research study; response; tumor; tumorigenesis; ubiquitin ligase

 DESCRIPTION (provided by applicant): Ubiquitin biology represents a major mode of cellular regulation. The final step of ubiquitination involves the recognition of a substrate by a "ubiquiti ligase". Humans encode over 700 such ligases, significantly exceeding the number of kinases encoded by the genome. Among these ubiquitin ligase genes are many oncogenes and tumor suppressors. In many cases the substrates of these ligases have not been well defined, in part due to a lack of robust technologies for identifying these substrates. We have developed a method, which we call "Ligase Trapping" which traps the nascent ubiquitinated substrate on the ubiquitin ligase and allows for its co-purification and identification. Here, we will apply this technology to a group of ubiquitin ligase specificity subunits called F box proteins. Human's encode about 80 such proteins, and we will focus on nine that have either been shown to be mutated or mis-expressed in tumors, or have targets whose functions have been linked to tumorigenesis (such as cell cycle regulators and DNA damage response proteins). In addition, we will use this technology to characterize the binding surfaces of these substrate binders. Together, these data will allow the identification of potential drug targets, and, more importantly will allow the generation of inhibitors of the F box proteins themselves in such a way as to affect the turnover of the most relevant substrates.

 描述（由申请人提供）：泛素生物学代表了细胞调节的主要模式。泛素化的最后一步涉及“泛素连接酶”对底物的识别，人类编码超过 700 种此类连接酶，远远超过了由“泛素连接酶”编码的激酶的数量。这些泛素连接酶基因中有许多癌基因和肿瘤抑制基因，在许多情况下，这些连接酶的底物尚未明确定义。部分原因是缺乏识别这些底物的可靠技术，我们开发了一种称为“连接酶捕获”的方法，该方法将新生的泛素化底物捕获在泛素连接酶上并允许其共纯化和识别。将这项技术应用于一组称为 F 盒蛋白的泛素连接酶特异性亚基，人类编码大约 80 种此类蛋白质，我们将重点关注 9 种已被证明发生突变的蛋白质。或在肿瘤中错误表达，或具有与肿瘤发生有关的功能（例如细胞周期调节因子和DNA损伤反应蛋白）。此外，我们将使用该技术来表征这些底物结合物的结合表面。这些数据将允许识别潜在的药物靶点，更重要的是，将允许生成 F 盒蛋白本身的抑制剂，从而影响 最相关底物的周转率。