Structure and Mechanism in intracellular Notch signaling

细胞内Notch信号传导的结构和机制

• 批准号: 7613390
• 负责人: DOUGLAS E. BARRICK
• 金额: $ 27.13万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7613390
• 关键词: Accounting; Adult; Affinity; Ankyrin Repeat; B-Lymphocytes; Binding; Biochemical; Biological Assay; Cell Communication; Cell Culture Techniques; Cell Fate Control; Cell Maintenance; Cell Surface Receptors; Cells; Code; Collaborations; Complex; Couples; Coupling; Critical Pathways; Crystallography; Cultured Cells; Data; Defect; Deletion Mutagenesis; Development; Disease; Drosophila genus; Drosophila melanogaster; Event; Exhibits; Foundations; Genes; Genetic; Genetic Transcription; Goals; Grant; Health; Homologous Gene; Human; Human Herpesvirus 4; Human Virus; In Vitro; Lead; Ligands; Link; Maintenance; Malignant Neoplasms; Mammalian Cell; Mammals; Mining; Molecular; Mutation; Notch Signaling Pathway; Organism; Pathway interactions; Pear; Peptides; Play; Population; Positioning Attribute; Production; Proteins; Reagent; Regulation; Research; Research Personnel; Resolution; Role; Signal Transduction; Specificity; Stem cells; Structure; Syndrome; Testing; Thermodynamics; Tissues; Ubiquitination; Universities; Viral; Work; base; cell type; computerized data processing; information processing; inhibitor/antagonist; insight; late disease onset; nervous system disorder; notch protein; precursor cell; progenitor; programs; receptor; stem cell biology; structural genomics; transcription factor; ubiquitin ligase; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): In humans and other multicellular organisms, complex tissues with different cell types must be formed from a uniform population of precursor cells. Cellular differentiation is critical in the development of a mature organism from a single cell, and in generating and replacing differentiated cells from stem cells in adults. Loss of control of differentiation can result in cancer. One pathway by which cell fate is determined is the "Notch" signaling pathway, a cell-cell communication pathway that relays information between neighboring cells (through a cell surface receptor protein called Notch) and processes this information inside the cell to tailor cell fate as needed. We are investigating the quantitative and structural mechanisms of Notch signaling. We wish to understand how Notch pathway proteins inside the signal-receiving cell control fate. We are studying two distinct Notch signaling processes, each controlled by a distinct cellular regulator (Deltex and CSL). Thermodynamic, biochemical, and high resolution structural analysis of the Deltex, CSL, the Notch receptor, and other effectors will be used to advance understanding of the molecular interactions underlying these cell fate decisions. We will determine how Deltex binds Notch, how it modifies Notch (via ubiquitination), and which receptor proteins pair with which Deltex proteins. Studies will reveal how bivalent recognition of CSL by Notch couples to increase binding strength, and how it allosterically turns this transcription factor on, and how a human virus exploits this pathway. We will test the functional significance of our findings through genetic studies in the fruit fly and transcription assays in cultured cells. We will take advantage of a Structural Genomics Consortium pipeline (University of Toronto) to increase the number of Notch proteins available for biophysical and biochemical analysis. Together, these studies will provide an understanding of Notch signaling that connects atomic structure to quantitative mechanism and function in the organism. Our research to better understand Notch signaling is important to human health, since defects in the Notch pathway lead to various forms of human cancer and to developmental malformities, and may ultimately contribute to treatment of these conditions. Our research is also important for understanding stem cells biology, which is maintained by Notch signaling.

描述（由申请人提供）：在人类和其他多细胞生物体中，具有不同细胞类型的复杂组织必须由统一的前体细胞群形成。细胞分化对于从单细胞发育成成熟生物体以及从成人干细胞产生和替换分化细胞至关重要。失去分化控制可能导致癌症。决定细胞命运的一条途径是“Notch”信号传导途径，这是一种细胞间通讯途径，可在相邻细胞之间传递信息（通过称为 Notch 的细胞表面受体蛋白），并在细胞内处理此信息以调整细胞命运：需要。我们正在研究 Notch 信号传导的定量和结构机制。我们希望了解信号接收细胞内的 Notch 通路蛋白如何控制命运。我们正在研究两种不同的 Notch 信号传导过程，每个过程都由不同的细胞调节因子（Deltex 和 CSL）控制。 Deltex、CSL、Notch 受体和其他效应器的热力学、生化和高分辨率结构分析将用于促进对这些细胞命运决定背后的分子相互作用的理解。我们将确定 Deltex 如何结合 Notch、它如何修饰 Notch（通过泛素化）以及哪些受体蛋白与哪些 Deltex 蛋白配对。研究将揭示Notch配对如何对CSL进行二价识别以增加结合强度，以及它如何以变构方式打开该转录因子，以及人类病毒如何利用该途径。我们将通过果蝇的遗传学研究和培养细胞的转录测定来测试我们的发现的功能意义。我们将利用结构基因组学联盟管道（多伦多大学）来增加可用于生物物理和生化分析的 Notch 蛋白的数量。总之，这些研究将提供对将原子结构与生物体定量机制和功能联系起来的 Notch 信号传导的理解。我们为更好地了解 Notch 信号传导而进行的研究对人类健康非常重要，因为 Notch 通路的缺陷会导致各种形式的人类癌症和发育畸形，并可能最终有助于治疗这些疾病。我们的研究对于理解干细胞生物学也很重要，干细胞生物学是由 Notch 信号传导维持的。