Mechanisms of protein degradation that control signal transduction by Ras-Raf-MEK-ERK

Ras-Raf-MEK-ERK 控制信号转导的蛋白质降解机制

基本信息

批准号：
10418701
负责人：
Claire de la Cova
金额：
$ 7.6万
依托单位：
UNIVERSITY OF WISCONSIN MILWAUKEE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-06-04 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10418701
关键词：
Address BRAF gene Benign Benign Melanocytic Nevus Biosensor CCNE1 gene CDK2 gene Caenorhabditis elegans Cell Cycle Cell Proliferation Cells Centers for Disease Control and Prevention (U.S.)Data Development Drug Design Drug resistance Engineering Event FBXW7 gene Feedback Future Gene Mutation Genes Genetic Screening Goals Growth Human In Vitro Knowledge Lead MAP2K1 gene MAPK3 gene MEKs Malignant Neoplasms Mediating Mission Modeling Mole the mammal Molecular Molecular Target Mutation NRAS gene Nevi and Melanomas Nevus Orthologous Gene Outcome Outcome Study Output Pathway interactions Persons Pharmaceutical Preparations Pharmacotherapy Phenotype Phosphoric Monoester Hydrolases Phosphorylation Phosphorylation Site Phosphotransferases Post-Translational Protein Processing Protein Kinase Proteins Proto-Oncogene Proteins B-raf Public Health RNA Interference RNA interference screen Ras/Raf Recurrent disease Regulation Relapse Research Role Scientific Advances and Accomplishments Signal Transduction Testing Ubiquitination United States Work anticancer research base cost design drug development effective therapy expectation in vivo inhibitor insight melanoma mutant novel therapeutic intervention novel therapeutics prevent protein degradation raf Kinases tumor ubiquitin-protein ligase

项目摘要

Project Summary This proposal aims to discover mechanistic insights into how signal transduction by the highly conserved Ras-Raf-MEK-ERK pathway is modulated by regulated protein degradation. In humans, mutation of the gene BRAF, encoding one of three human Rafs, is a driver event in the development of malignant melanoma. Indeed, the mutation BRAF(V600E) confers constitutive kinase activation and is one of the most frequent mutations found in this cancer. Melanoma growth can be suppressed by inhibition of Raf kinase activity. However, development of drug resistance and disease relapse occurs frequently, indicating that truly effective treatments will require additional drug strategies. Our work, utilizing Caenorhabditis elegans as a simple model to investigate signal transduction by this conserved pathway, demonstrates that C. elegans Raf signaling is kept in check by inhibitory mechanisms that include phosphorylation and protein degradation (de la Cova and Greenwald, 2012). For this proposed research, we use the C. elegans model to design and begin an RNAi screen to identify conserved kinases and phosphatases that regulate Raf. We find that CDK2 is necessary for Raf protein degradation, and that loss of CDK2 enhances phenotypes caused by a mutant, activated Raf engineered to carry the same mutation as BRAF(V600E). Our long-term goal is to identify new molecular targets for the development of drugs capable of inhibiting Raf signaling. Toward this goal, our overall objectives in this application are to (i) identify additional kinases and phosphatases that impact Raf protein degradation and their requirement for regulating BRAF in human cells, and (ii) determine the mechanism and consequences of Raf regulation by the kinase CDK2. Our central hypothesis is that conserved mechanisms, such as phosphorylation of the Raf protein, act to prevent aberrant Raf activation. The rationale for this project is that our discovery of inhibitory control mechanisms governing signal transduction by the Ras-Raf-MEK-ERK pathway will inform new strategies of drug therapy for melanoma.

项目概要该提案旨在发现高度信号如何转导的机制见解保守的 Ras-Raf-MEK-ERK 通路受调节的蛋白质降解调节。在人类中，突变编码三种人类 Raf 之一的基因 BRAF 的突变是恶性疾病发展的驱动事件。黑色素瘤。事实上，突变 BRAF(V600E) 赋予组成型激酶激活作用，并且是最常见的突变之一。这种癌症中经常发现突变。抑制 Raf 激酶可抑制黑色素瘤的生长活动。然而，耐药性的产生和疾病复发的频繁发生，表明真正的有效的治疗需要额外的药物策略。我们的工作利用秀丽隐杆线虫作为研究这一保守途径信号转导的简单模型表明，线虫 Raf 信号传导受到抑制机制的控制，包括磷酸化和蛋白质降解（de 拉科瓦和格林沃尔德，2012）。对于这项拟议的研究，我们使用秀丽隐杆线虫模型来设计并开始 RNAi 筛选可识别调节 Raf 的保守激酶和磷酸酶。我们发现CDK2是 Raf 蛋白降解所必需的，CDK2 的缺失会增强突变体引起的表型，激活的 Raf 被设计为携带与 BRAF(V600E) 相同的突变。我们的长期目标是寻找新的开发能够抑制 Raf 信号传导的药物的分子靶标。为了这个目标，我们的本申请的总体目标是 (i) 识别影响 Raf 的其他激酶和磷酸酶蛋白质降解及其在人类细胞中调节 BRAF 的要求，以及 (ii) 确定激酶 CDK2 调节 Raf 的机制和后果。我们的中心假设是保守的机制，例如 Raf 蛋白的磷酸化，可以防止异常的 Raf 激活。该项目的基本原理是我们发现了控制信号的抑制控制机制 Ras-Raf-MEK-ERK 通路的转导将为黑色素瘤药物治疗的新策略提供信息。