Regulators of Tumorigenesis

肿瘤发生的调节因子 项目总结 RAS GTP酶代表了一个难以靶向的癌症驱动关系，突显了 需要更好地表征支配其基因的RAS相互作用生物分子的光谱 功能。使用旨在寻找以前遗漏的交互作用的新方法，最新的CA142635 努力定义了RAS RNA和蛋白质相互作用，以确定一个重要的功能 RAS的新的RNA结合能力以及38个与癌症相关的RAS相互作用蛋白。 RAS选择性和直接结合C/D盒小非编码RNA snoRNA，包括 SNORD50A/B，它有效地抑制RAS，并在高频共生缺失 人类癌症中存在RAS基因突变。活细胞邻近蛋白标记和质量分析 光谱检测到先前研究中遗漏的蛋白质相互作用，其中包括mTOR 和推测的棕榈酰基转移酶ZDHHC5。这一相互竞争的续约将定义 这些新的RAS相互作用的RNA和蛋白质在RAS驱动的肿瘤发生中的作用。 首先，我们将进一步表征SNORD50A/B snoRNA对RAS的影响及其作用 其他新发现的RAS结合的snoRNAs。基于SNORD50A/B的发现 SnoRNAs阻断法尼基转移酶与RAS的结合，我们将定义 Ras蛋白相互作用的SNORD50A/B缺失进一步揭示新的 发现了RNA对RAS的调节作用。我们还将系统地测试它的功能 RAS驱动的肿瘤上的其他RAS结合RNA，从6个snoRNA开始，它们是 在人类癌症中以高频率反复突变。Aim我将检验假设 SNORD50A/B snoRNAs重塑RAS蛋白相互作用组以调节RAS信号转导和 额外的RAS结合的snoRNAs改变了RAS在肿瘤发生中的作用。 其次，我们将描述新发现的RAS-mTOR在肿瘤发生中的相互作用 并将确定其他邻近蛋白对致癌RAS的功能重要性 功能。我们将定义RAS亚型上的结构域，并对观察到的mTOR负责 RAS与mTOR的直接结合，研究RAS-mTOR结合对mTORC1vs 并评估破坏RAS-mTOR结合对RAS-mTOR活性的影响。 体内驱动的肿瘤形成。我们还将有系统地删除38个新确定的RAS 并量化它们对RAS驱动的肿瘤发生的影响。AIM II将测试 RAS-mTOR结合在RAS驱动的肿瘤发生中起作用的假说 多种新发现的RAS相互作用蛋白影响肿瘤中RAS信号转导结果。 在拟议供资周期结束时，这项工作将界定新的 在RAS驱动的肿瘤发生中发现了RAS结合的RNA和RAS近端蛋白。 1

REGULATORS OF TUMORIGENESIS PROJECT SUMMARY Ras GTPases represent a cancer-driving nexus that is difficult to target, underscoring the need to better characterize the spectrum of Ras-interacting biomolecules that govern its function. Using new methods designed to find previously missed interactors, recent CA142635 efforts defined the Ras RNA and protein interactomes to identify a functionally important new RNA binding capacity for Ras as well as 38 new cancer-relevant Ras-interacting proteins. Ras selectively and directly bound C/D box small non-coding RNAs snoRNAs, including SNORD50A/B, which potently inhibited Ras and was deleted at high frequency co-occurantly with RAS gene mutation in human cancer. Live-cell vicinal protein labeling followed by mass spectrometry detected protein interactions missed in prior studies, among which were mTOR and the putative palmitoyltransferase, ZDHHC5. This competing renewal will define the function of these new Ras interacting RNA and proteins in Ras-driven tumorigenesis. First, we will further characterize SNORD50A/B snoRNA impacts on Ras as well as the role of other newly discovered Ras-binding snoRNAs. Based on the finding that SNORD50A/B snoRNAs block farnesyltransferase binding to Ras, we will define the global impact of SNORD50A/B deletion on Ras protein-protein interactions to shed further light on the newly discovered modulation of Ras by RNA. We will also systematically test the function of additional Ras-binding RNAs on Ras-driven neoplasia, beginning with 6 snoRNAs that are recurrently mutated at high frequency in human cancer. Aim I will test the hypotheses that SNORD50A/B snoRNAs remodel the Ras protein interactome to regulate Ras signaling and that additional Ras-binding snoRNAs alter Ras action in tumorigenesis. Second, we will characterize the newly identified Ras-mTOR interaction in tumorigenesis and will define the functional importance of additional vicinal proteins to oncogenic Ras function. We will define the domains on Ras isoforms and mTOR responsible for the observed direct binding of Ras to mTOR, examine the impact of Ras-mTOR binding on mTORC1 vs mTORC2 kinase activity and assess the impacts of disrupting Ras-mTOR binding on Ras- driven tumorigenesis in vivo. We will also systematically delete 38 newly identified Ras proximal proteins and quantify their impact on Ras-driven tumorigenesis. Aim II will test the hypotheses that Ras-mTOR binding is functional in Ras-driven tumorigenesis and that multiple newly identified Ras-interacting proteins impact Ras signaling outcomes in neoplasia. At the end of the proposed funding cycle, this effort will define the function of newly identified Ras-binding RNAs and Ras-proximal proteins in Ras-driven tumorigenesis. 1