Structure and Mechanism in intracellular Notch signaling

细胞内Notch信号传导的结构和机制

• 批准号: 7804492
• 负责人: DOUGLAS E. BARRICK
• 金额: $ 26.78万
• 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 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7804492
• 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中，必须由统一的前体细胞形成具有不同细胞类型的复杂组织。细胞分化对于从单个细胞的成熟生物的发展以及产生和代替成人干细胞分化细胞的成熟生物至关重要。失去分化的控制可能导致癌症。确定细胞命运的一个途径是“ Notch”信号通路，这是一种细胞 - 细胞通信途径，它在相邻细胞之间（通过称为Notch的细胞表面受体蛋白）中继信息，并在细胞内处理这些信息以根据需要定制细胞命运。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种细胞控制命运中的Notch途径蛋白如何。我们正在研究两个不同的Notch信号传导过程，每个过程都由不同的细胞调节剂（Deltex和CSL）控制。 Deltex，CSL，Notch受体和其他效应子的热力学，生化和高分辨率结构分析将用于提高对这些细胞命运决定基础的分子相互作用的理解。我们将确定Deltex如何结合Notch，如何修饰Notch（通过泛素化）以及哪种受体蛋白与哪些Deltex蛋白配对。研究将揭示Notch夫妇对CSL的认识，以提高结合强度，以及它如何变构开启该转录因子，以及人类病毒如何利用这一途径。我们将通过在培养细胞中的果蝇和转录测定中的遗传研究来检验发现的功能意义。我们将利用结构性基因组学财团（多伦多大学）来增加可用于生物物理和生化分析的Notch蛋白数量。这些研究将共同​​了解将原子结构与生物体中定量机制和功能联系起来的Notch信号传导。我们以更好地理解Notch信号的研究对人类健康很重要，因为Notch途径中的缺陷导致各种形式的人类癌症和发育畸形，并最终可能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，这是通过Notch信号传导维持的。