Understanding the function of microtubule-associated signalling networks in health and disease: role of adaptor proteins in network assembly

了解微管相关信号网络在健康和疾病中的功能：接头蛋白在网络组装中的作用

• 批准号: 2599452
• 金额: --
• 依托单位: University of Liverpool
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2599452
• 关键词: Understanding; function; microtubule; associated; signalling

Microtubules (MTs) are essential for the regulation of key processes in eukaryotic cells, in particular cargo transport, cell division and migration. At the heart of these cellular processes are dynamic assemblies of MT plus-end tracking proteins (+TIPs). They guide MTs precisely to different areas in cells, and then selectively trigger specific responses "on arrival". +TIP-network assembly is governed by adaptor proteins that contain a small set of short protein interaction motifs (SxIP, EEY and basic-S/P-), embedded into intrinsically disordered regions however, this story is far from complete. Most of the research is focused on individual components, and little quantitative analysis is conducted. As a result, predictions from current models are unreliable. This project is an integral part of an on-going study to develop a comprehensive, multi-component model of +TIP signalling networks. Mutations of adaptor proteins in cancer contributes to disease progression and metastasis, demonstrating their critical role and the justification for more comprehensive research. Using NMR, X-ray crystallography and ITC we will solve structures of the adaptor protein complexes and define thermodynamic characteristics of the main interactions within the plus-end networks that are currently uncharacterised. Primarily we will focus on the SxIP, EEY and basic-S/P-motifs that interconnect the core of the networks. We will also monitor the networks in live cells with high-resolution fluorescent microscopy and use cell-permeable structure-based peptide inhibitors to modulate their characteristics by disrupting specific interactions. These changes will be correlated with the effects on migration and proliferation of normal and cancer cells to define the contributions of specific proteins. With engineered cell substrates we will modulate cell environment to emulate healthy and pathological tumour and ageing tissues, thus defining the ability of cells to react to the environmental changes. Joining all the information together we hope to identify critical components of the signalling networks and test their potential as drug targets.

微管 (MT) 对于真核细胞关键过程的调节至关重要，特别是货物运输、细胞分裂和迁移。这些细胞过程的核心是 MT 加末端跟踪蛋白 (+TIP) 的动态组装。它们将 MT 精确引导到细胞中的不同区域，然后选择性地“到达”时触发特定反应。 +TIP 网络组装由接头蛋白控制，接头蛋白包含一小组短蛋白相互作用基序（SxIP、EEY 和 basic-S/P-），嵌入本质上无序的区域，但是，这个故事还远未完成。大多数研究都集中在单个组成部分，很少进行定量分析。因此，当前模型的预测是不可靠的。该项目是正在进行的研究的一个组成部分，该研究旨在开发+TIP 信号网络的综合、多组件模型。癌症中接头蛋白的突变有助于疾病进展和转移，证明了它们的关键作用以及进行更全面研究的合理性。利用 NMR、X 射线晶体学和 ITC，我们将解析接头蛋白复合物的结构，并定义目前尚未表征的正端网络内主要相互作用的热力学特征。我们将主要关注互连网络核心的 SxIP、EEY 和基本 S/P 基序。我们还将用高分辨率荧光显微镜监测活细胞中的网络，并使用基于细胞可渗透结构的肽抑制剂通过破坏特定的相互作用来调节其特性。这些变化将与对正常细胞和癌细胞迁移和增殖的影响相关联，以确定特定蛋白质的贡献。通过工程细胞基质，我们将调节细胞环境以模拟健康和病理肿瘤和衰老组织，从而定义细胞对环境变化做出反应的能力。将所有信息结合在一起，我们希望识别信号网络的关键组成部分并测试它们作为药物靶点的潜力。