Project 3: Defining and targeting mechanisms of E2F transcription factor regulation

项目3：E2F转录因子调控的定义和靶向机制

基本信息

批准号：
10332382
负责人：
Seth Michael Rubin
金额：
$ 27.65万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-25 至 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10332382
关键词：
Acetylation Adaptor Signaling Protein Address Affinity Architecture Binding Biochemical Biochemistry Cancer Model Cell Cycle Cell Cycle Progression Cell Cycle Regulation Cell Proliferation Cell division Cells Cellular Assay Cellular biology Chemicals Chromatin Collaborations Complex Consensus Crystallization Cullin Proteins Cyclin-Dependent Kinases Cyclins DNA DNA biosynthesis Data Defect Dependence E2F transcription factors Enzymes Event Foundations G1/S Transition Gene Activation Gene Expression Genes Genetic Genetic Transcription Genomic approach Goals Growth Hydrophobicity Interphase Cell Knowledge Malignant Neoplasms Mediating Mitosis Modification Molecular Molecular Analysis Mutagenesis Nucleosomes Pathway interactions Phenotype Phosphorylation Post-Translational Protein Processing Post-Translational Regulation Protein Tyrosine Kinase Proteins Proteomics Regulation Repression Resolution Retinoblastoma Protein Route S phase Site Specificity Structure Substrate Cycling Substrate Interaction Testing Therapeutic Time Tumor Suppressor Proteins base cancer cell cell growth chromatin protein chromatin remodeling cyclin F design inhibitor molecular imaging neoplastic cell novel novel therapeutic intervention programs promoter protein function protein protein interaction reconstitution recruit refractory cancer structural biology therapeutically effective ubiquitin-protein ligase

项目摘要

PROJECT SUMMARY Activation of E2F transcription factors is the most downstream event in the retinoblastoma protein (Rb) tumor suppressor pathway, which controls entry into the cell cycle and proliferation. E2F stimulates expression of genes needed for DNA synthesis during S phase and for further progression into mitosis. Inactivation of Rb and aberrant activation of the E2F-controlled transcription program is a hallmark of cancer. While Rb repression of E2F has been extensively characterized, little is known regarding how E2F activates transcription and how additional regulatory mechanisms control E2F function. Here we explore the structural mechanisms underlying the function and posttranslational regulation of E2F. Our first aim is to determine how E2F associates with chromatin and influences chromatin architecture to induce gene expression. We will use structural, biochemical, and cellular assays to investigate how E2F binds nucleosomes and how those interactions are critical for modulating chromatin architecture as cells activate the transcription program for S phase entry. Our second aim is to identify novel E2F posttranslational modifications and how modifications including phosphorylation and acetylation modulate E2F association with chromatin and co-activator proteins. Our third aim is to reveal how E2F proteins are recognized by the E3 ligase adaptor protein Cyclin F (CycF) for ubiquitylation and degradation. A structural and cell biology approach will be applied to define the CycF-binding motif, and we will test the hypothesis that chemical inhibition of CycF can deregulate the cell cycle in model cancer cells. In collaboration with the other program projects and cores, our study will provide unprecedented molecular analysis of E2F function and regulation. Our experimental results will provide foundational knowledge that can be applied to design new therapeutic strategies that target E2F activity in diverse cancers.

项目概要 E2F 转录因子的激活是视网膜母细胞瘤蛋白 (Rb) 肿瘤中最下游的事件抑制途径，控制进入细胞周期和增殖。 E2F 刺激表达 S 期 DNA 合成和进一步进入有丝分裂所需的基因。 Rb 失活 E2F 控制的转录程序的异常激活是癌症的一个标志。而铷 E2F 的抑制已被广泛表征，但关于 E2F 如何激活转录知之甚少以及额外的监管机制如何控制 E2F 功能。在这里我们探讨一下结构机制 E2F 的功能和翻译后调节的基础。我们的首要目标是确定 E2F 如何与染色质结合并影响染色质结构诱导基因表达。我们将使用结构、生化和细胞测定来研究 E2F 如何结合核小体以及这些相互作用如何在细胞激活染色质结构时对调节染色质结构至关重要进入S期的转录程序。我们的第二个目标是识别新颖的 E2F 翻译后修饰以及包括磷酸化和乙酰化在内的修饰如何调节 E2F 与染色质的关联以及共激活蛋白。我们的第三个目标是揭示 E3 连接酶适配器如何识别 E2F 蛋白蛋白质 Cyclin F (CycF) 用于泛素化和降解。结构和细胞生物学方法将是应用于定义 CycF 结合基序，我们将测试 CycF 的化学抑制可以放松模型癌细胞中细胞周期的调节。与其他计划项目和核心合作，我们的研究将提供前所未有的分子 E2F功能和调节分析。我们的实验结果将提供基础知识可用于设计针对不同癌症中 E2F 活性的新治疗策略。