Regulation of Ras in development by post-translational modification

通过翻译后修饰调控 Ras 的发育

基本信息

批准号：
10356918
负责人：
CATHIE M PFLEGER
金额：
$ 33.9万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-13 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10356918
关键词：
Address Alleles Biochemistry Biological Cell Proliferation Cell Survival Cells Complement Cues Development Developmental Biology Drosophila genus Elements Embryonic Development Ensure Feedback Genetic Goals Homeostasis Impairment In Vitro Lead Longevity Malignant Neoplasms Mediating Modification Molecular Mutation N-terminal Neurofibromatosis 1 Noonan Syndrome Output Pathway interactions Pattern Phenotype Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Post-Translational Protein Processing Process Proteins Ras Signaling Pathway Regulation Research Role Signal Pathway Signal Transduction System Testing Therapeutic Translating Tumor Suppression Tyrosine Tyrosine Phosphorylation Ubiquitin Ubiquitination Up-Regulation Work developmental disease in vivo inorganic phosphate insight mutant novel response screening trafficking

项目摘要

SUMMARY Eukaryotic signaling pathways translate external cues in specific contexts into diverse biological outputs. Proper timing and duration of a signaling response requires careful regulation of stability, trafficking, activation, and deactivation of multiple components within a network. This overall coordination can involve positive feedback circuits to amplify signaling and/or negative feedback circuits to constrain signaling. The Ras signaling pathway is crucial in development to regulate proliferation, cell survival, cell fate, and patterning. Mutations in Ras or other components of the pathway that lead to increased Ras signaling cause developmental disorderes collectively known as “Rasopathies” such as Noonan's syndrome or Neurofibromatosis Type 1 (NF1). Using Drosophila, we discovered that Rabex-5 (an E3) downregulates Ras by promoting its mono- and di-ubiquitination. Specifically, we discovered that impairing Ras ubiquitination in vivo in Drosophila led to striking effects on cell proliferation, cell survival, developmental patterning, and organismal longevity, reflecting a role for Ras ubiquitination in development, tumor suppression, and survival. The power of Drosophila genetics and the well-established paradigm of studying Ras in Drosophila make this system ideally suited to characterize this phenomenon in a multi-cellular context. We more recently discovered that Rabex-5 targeting of Ras requires an N-terminal tyrosine in Ras, Tyrosine 4 (Y4). We hypothesize that phosphorylation of this tyrosine directs targeting of Ras by Rabex-5. Our hypothesis predicts that the kinase(s) and phosphatase(s) that regulate modification of Y4 are crucial regulators of Ras signaling. The goal of our application is to elucidate the role of tyrosine phosphorylation of Ras to direct its inhibitory Rabex-5- mediated ubiquitination and maintain pathway homeostasis. Specifically, we propose to further characterize the biological role of Y4 phosphorylation in Ras ubiquitination, to identify the Ras Y4 kinase(s) and phosphatase(s), and to characterize their roles in regulating Ras in development.

概括真核信号通路将特定环境中的外部线索转化为潜水员生物产量。信号反应的适当时间和持续时间需要仔细调节稳定性，运输，激活，以及网络中多个组件的停用。这种整体协调可能涉及积极反馈电路以扩大信号传导和/或负反馈电路以限制信号传导。拉斯信号通路对于调节增殖，细胞存活，细胞命运和图案的发展至关重要。 RAS中的突变或途径的其他组件导致RAS信号增加的原因发育障碍统称为“ rasopathies”，例如诺南综合症或 1型神经纤维瘤病（NF1）。使用果蝇，我们发现Rabex-5（A e3）下调了RAS 通过促进其单次泛素化。具体来说，我们发现在果蝇中的体内对细胞增殖，细胞存活，发育模式和生物体的寿命，反映了RAS泛素化在发育，肿瘤抑制和生存中的作用。果蝇遗传学的力量和果蝇研究RAS的公认范式使得系统非常适合在多细胞上下文中表征这种现象。我们最近发现 RAS的Rabex-5靶向RAS需要在RAS中进行N末端酪氨酸，酪氨酸4（Y4）。我们假设这一点这种酪氨酸的磷酸化指导Rabex-5靶向RAS。我们的假设预测调节Y4修饰的激酶（S）和磷酸酶是RAS信号传导的关键调节剂。目标我们的应用是阐明Ras酪氨酸磷酸化的作用，以指导其抑制性Rabex-5-- 介导的泛素化并保持途径稳态。具体而言，我们建议进一步表征 Y4磷酸化在RAS泛素化中的生物学作用，以鉴定Ras Y4激酶和磷酸酶（S），并表征其在调节RAS发育中的作用。