Elucidation of Novel Post-Translational Mechanisms That Regulate Notch Activity

阐明调节Notch活性的新型翻译后机制

• 批准号: 8524194
• 负责人: Ryan Scott Underwood
• 金额: $ 4.15万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8524194
• 关键词: Acute T Cell Leukemia; Animal Model; Binding; Caenorhabditis elegans; Cell Nucleus; Cleaved cell; Complex; DNA; DNA-Binding Proteins; Defect; Development; Disease; Drosophila genus; Genetic; Genetic Techniques; Genetic Transcription; Goals; Human; Investigation; Laboratories; Lead; Ligands; Link; Lobular Neoplasia; MAP Kinase Gene; Malignant Neoplasms; Mammals; Methods; Microscopy; Mitogen-Activated Protein Kinases; Mutation; Notch Signaling Pathway; Oncogenic; Organism; Pathway interactions; Pattern; Proteins; Refractory; Regulation; Reporter; Research; Role; Signal Transduction; System; Testing; Time; Tissues; Transcription Coactivator; Work; base; design; gain of function; genetic regulatory protein; human disease; in vivo; information gathering; notch protein; novel; novel therapeutics; prevent; promoter; public health relevance; receptor; transcription factor

DESCRIPTION (provided by applicant): The misregulation of Notch signaling has been implicated in developmental defects in many organisms and in human diseases, most notably T-Cell Acute Lymphoblastic Leukemia (T-ALL). This research will characterize the regulation of activated forms of Notch in developmental contexts using Caenorhabditis elegans. Specifically, this research will examine two potentially novel forms of negative regulation that are not understood at the mechanistic level. Both forms of regulation block C. elegans LIN-12/Notch function in the nucleus. In one case, crosstalk with the Ras-MAP kinase pathway blocks the effect of constitutively active LIN-12/Notch specifically where Ras-MAP kinase is activated. In the second case, the novel DNA-binding protein, LIN-14, which regulates developmental timing, blocks constitutively active LIN-12/Notch in a particular tissue. The constitutively active forms o LIN-12 blocked is analogous to mutations found in certain cancers such as T-ALL. In the course of this research, I will characterize the regulation of two required Notch accessory proteins, a well-conserved transcription factor (LAG-1) and a coactivator (SEL-8), and the transcriptional complex they form with LIN-12. Additionally, these methods will be expanded to characterize additional forms of regulation found through manipulation of LIN-12 target reporters. By characterizing these two mechanisms, this proposal aims to increase the understanding of how the critical Notch pathway is regulated and potentially provide novel therapeutic strategies for diseases such as T-ALL.

描述（由申请人提供）：Notch 信号传导的失调与许多生物体和人类疾病的发育缺陷有关，最显着的是 T 细胞急性淋巴细胞白血病 (T-ALL)。这项研究将利用秀丽隐杆线虫来描述Notch激活形式在发育环境中的调节。具体来说，这项研究将研究两种潜在的新颖的负调控形式，这些形式在机械层面上尚未被理解。两种形式的调节块秀丽隐杆线虫 LIN-12/Notch 在细胞核中发挥作用。在一种情况下，与 Ras-MAP 激酶通路的串扰会阻断组成型活性 LIN-12/Notch 的作用，特别是在 Ras-MAP 激酶被激活的情况下。在第二种情况下，调节发育时间的新型 DNA 结合蛋白 LIN-14 会阻断特定组织中持续活跃的 LIN-12/Notch。 LIN-12 被阻断的组成型活性形式类似于在某些癌症（如 T-ALL）中发现的突变。在这项研究过程中，我将描述两种必需的 Notch 辅助蛋白（一种高度保守的转录因子 (LAG-1) 和一种共激活因子 (SEL-8)）的调节，以及它们与 LIN-12 形成的转录复合物。此外，这些方法将得到扩展，以表征通过操纵 LIN-12 目标报告基因发现的其他形式的监管。通过表征这两种机制，该提案旨在加深对关键 Notch 通路如何调节的理解，并可能为 T-ALL 等疾病提供新的治疗策略。