Molecular dissection of Oncoprotein-induced apoptosis

癌蛋白诱导细胞凋亡的分子解剖

• 批准号: 6954112
• 负责人: GERARD IAN EVAN
• 金额: $ 31.06万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6954112
• 关键词: apoptosis; genetic models; genetically modified animals; laboratory mouse; mitochondria; molecular oncology; oncoproteins; posttranslational modifications; protein protein interaction; protein structure function; proteolysis; proteomics; tissue /cell culture; transcription factor; transfection

DESCRIPTION (provided by applicant): The induction of apoptosis by the Myc oncoprotein is considered to be a major innate mechanism preventing the emergence of neoplasia. However, although many, diverse proteins have each been proposed as key downstream apoptotic effectors, the molecular mechanisms linking Myc with the basal the apoptotic machinery are largely unknown. Myc sensitizes cells to apoptosis induced by a wide range of insults, suggesting that Myc acts to prime some central determinant of apoptotic decision-making. Consistent with this, our work has directly implicated the mitochondrion as rapid and immediate target of Myc pro-apoptotic action. Based upon this observation, we have used a biased proteomic approach to identify Bcl-xL-interacting proteins that rapidly change in abundance or mobility following acute Myc activation in human and rodent fibroblasts. Three candidates have been identified - the known pro-apoptotic BH3 protein Bim, a novel protein with a consensus BH3 domain called PDCD10 recently shown to be encoded by a Myc target gene, and the small G protein Rab7. We propose to validate each of these candidate Myc apoptotic effectors by a combination of molecular, cell and molecular biological and genetics strategies, both in vitro and in several tissues in vivo using a unique set of switchable Myc mouse transgenic models. Finally, we propose to extend our initial proteomic study to establish a map of the proteomic changes in mitochondria that rapidly accompany Myc activation, with particular emphasis on the restricted proteins in the outer mitochondrial compartment most likely targeted by Myc apoptotic effectors. These studies will provide new insights into the molecular machinery linking deregulated proliferation with its innate tumor suppressive apoptotic failsafe, and how that linkage is severed in cancer cells.

描述（由申请人提供）：Myc癌蛋白诱导细胞凋亡被认为是防止瘤形成出现的主要先天机制。然而，尽管许多不同的蛋白质都被认为是关键的下游凋亡效应子，但将 Myc 与基础凋亡机制联系起来的分子机制在很大程度上还是未知的。 Myc 使细胞对多种损伤诱导的细胞凋亡敏感，这表明 Myc 可以启动细胞凋亡决策的一些核心决定因素。与此一致，我们的工作直接表明线粒体是 Myc 促凋亡作用的快速且直接的目标。基于这一观察，我们使用有偏向的蛋白质组学方法来鉴定 Bcl-xL 相互作用蛋白，这些蛋白在人类和啮齿类成纤维细胞中 Myc 急性激活后丰度或迁移率迅速变化。已经确定了三个候选蛋白——已知的促凋亡 BH3 蛋白 Bim（一种具有 BH3 共有结构域的新型蛋白，称为 PDCD10，最近显示由 Myc 靶基因编码）和小 G 蛋白 Rab7。我们建议通过分子、细胞和分子生物学和遗传学策略的组合，使用一组独特的可切换 Myc 小鼠转基因模型在体外和体内多个组织中验证每个候选 Myc 凋亡效应器。最后，我们建议扩展我们最初的蛋白质组学研究，以建立伴随 Myc 激活快速发生的线粒体蛋白质组变化图谱，特别强调线粒体外区室中最有可能被 Myc 凋亡效应子靶向的限制性蛋白质。这些研究将为将失调的增殖与其固有的肿瘤抑制性细胞凋亡故障保护联系起来的分子机制，以及如何在癌细胞中切断这种联系提供新的见解。