Combining structural, biophysical and in vivo techniques to probe structure/function of stem cell signalling receptor, Notch

结合结构、生物物理和体内技术来探测干细胞信号受体Notch的结构/功能

• 批准号: 2898935
• 金额: --
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2898935
• 关键词: Combining; structural; biophysical; vivo; techniques

important challenge in biological research today. This is exemplified by studies of mutations in the Notch gene, an important signalling receptor regulating many aspects of development and stem cell activity. Notch is activated by binding of ligand to the extracellular domain which is comprised of numerous tandem repeats related to Epidermal Growth Factor (EGF). Ligand binding to the EGF region initiates proteolytic cleavages that release the Notch intracellular domain (NICD) to translocate to the nucleus, where NICD interacts with transcription factors and coactivator proteins to switch on target gene expression. The ankyrin domain region within the NICD plays an important role, providing binding sites for different components of the nuclear complex. The ankyrin region also forms a regulatory platform for binding components such as ubiquitin ligases and Adapter protein complexes involved in the endocytic trafficking and sorting which play an important part in modulating Notch signalling levels. Drosophila genetic/developmental analysis and cancer genome sequencing has shown that point mutations in different locations of the Notch protein including different EGF and Ankyrin repeats produce a diversity of phenotypic outcomes. The mechanistic basis, i.e. how the sequence change leads to phenotypic outcome through altered Notch function, is poorly understood for many of these mutations but the diversity of outcomes suggests different mutations probe distinct regulatory interactions.

这是当今生物学研究的一个重大挑战。Notch基因突变的研究就是例证，Notch基因是一种重要的信号受体，调节发育和干细胞活性的许多方面。Notch通过配体与细胞外结构域的结合而被激活，所述细胞外结构域由与表皮生长因子（EGF）相关的许多串联重复序列组成。配体与EGF区域的结合启动蛋白水解裂解，释放Notch胞内结构域（NICD）以易位至细胞核，在细胞核中NICD与转录因子和共激活蛋白相互作用以开启靶基因表达。NICD内的锚蛋白结构域区域起着重要作用，为核复合物的不同组分提供结合位点。锚蛋白区域还形成了用于结合组分的调节平台，所述结合组分例如参与内吞运输和分选的泛素连接酶和衔接子蛋白复合物，其在调节Notch信号传导水平中起重要作用。果蝇遗传/发育分析和癌症基因组测序表明，Notch蛋白不同位置的点突变，包括不同的EGF和锚蛋白重复序列，产生多种表型结果。机制基础，即序列变化如何通过改变Notch功能导致表型结果，对于许多这些突变知之甚少，但结果的多样性表明不同的突变探针不同的调控相互作用。