Deciphering the ubiquitin code in stress signaling and membrane trafficking

破译应激信号传导和膜运输中的泛素代码

• 批准号: 10330680
• 负责人: Jason A MacGurn
• 金额: $ 39.63万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10330680
• 关键词: Address; Amino Acids; Binding Proteins; Biochemical; Biology; Cell physiology; Cells; Cellular Stress Response; Code; Elements; Enzymes; Eukaryotic Cell; Family; Genetic; Glucose Transporter; Human; Investigation; Length; Malignant Neoplasms; Membrane; Modification; Nerve Degeneration; Pathway interactions; Peptides; Phosphorylation; Phosphotransferases; Polymers; Positioning Attribute; Post-Translational Protein Processing; Proteins; Proteomics; Quality Control; Regulation; Research; SLC2A1 gene; Signal Transduction; Stress; Syndrome; Ubiquitin; Work; Writing; Yeasts; base; biological adaptation to stress; blood glucose regulation; cancer type; human disease; insight; protein degradation; protein protein interaction; proteostasis; response; therapeutic target; trafficking

Project Summary Ubiquitin is a 76 amino acid peptide that can be covalently conjugated to substrates to alter protein fate in diverse ways, regulating protein degradation, trafficking, subcellular localization and protein-protein interactions. Given its versatility, ubiquitin regulates many fundamental cellular processes, and its dysregulation is associated with many human diseases ranging from neurodegeneration to cancer. Ubiquitin networks include conjugating and deconjugating enzymes as well as effector pathways comprised of ubiquitin binding proteins that direct the fate of ubiquitin-modified substrates. All of these elements work together to “write”, “read”, and “edit” the ubiquitin code – which ultimately consists of ubiquitin polymers of different lengths and topologies that determine which effector pathways are engaged. Here, we describe two main research directions that will result in a deeper understanding of the ubiquitin code and how it regulates diverse cellular functions, including stress signaling and membrane trafficking. The first research direction will address how phosphorylation of ubiquitin at the Ser57 position regulates stress responses in yeast and human cells. The proposed studies will build on our recent discovery of a small group of Ser57 ubiquitin kinases conserved from yeast to humans and will include genetic, biochemical, and proteomic approaches. Specifically, we will determine how these kinases and Ser57 phosphorylation of ubiquitin contribute to the cellular stress response, and we will address how ubiquitin phosphorylation alters its interaction profile and engagement with effector pathways. This research will contribute transformative new insights into the biology of ubiquitin and proteostasis. The second research direction will address how human glucose transporters are regulated by ubiquitin modification and endocytic trafficking. Glucose transporters of the GLUT family are key regulators of cellular glucose homeostasis, and yet regulation of their trafficking and quality control remain poorly characterized. Here, we describe lines of investigation based on our recent findings that GLUT1 endocytic trafficking is regulated by specific ubiquitin modifications. These studies have important implications for cellular glucose homeostasis and human diseases including GLUT1 Deficiency Syndrome and many types of cancer. Together, these research directions will result in a deeper understanding of the ubiquitin code, membrane trafficking, and stress responses in eukaryotic cells.

项目概要 泛素是一种 76 个氨基酸的肽，可以与底物共价结合以改变蛋白质的命运 多种方式，调节蛋白质降解、运输、亚细胞定位和蛋白质-蛋白质 互动。鉴于其多功能性，泛素调节许多基本的细胞过程，并且它 失调与许多人类疾病有关，从神经退行性疾病到癌症。泛素 网络包括缀合和解缀酶以及由泛素组成的效应途径 指导泛素修饰底物命运的结合蛋白。所有这些元素共同作用 “写入”、“读取”和“编辑”泛素代码——最终由不同长度的泛素聚合物组成 以及决定参与哪些效应器通路的拓扑。在这里，我们描述两个主要研究 方向将导致更深入地了解泛素代码及其如何调节不同的细胞 功能，包括应激信号传导和膜运输。第一个研究方向将解决如何 泛素 Ser57 位点的磷酸化可调节酵母和人类细胞的应激反应。这 拟议的研究将建立在我们最近发现的一小群 Ser57 泛素激酶的基础上，这些激酶保守于 酵母对人类的影响，将包括遗传、生化和蛋白质组学方法。具体来说，我们将 确定这些激酶和泛素 Ser57 磷酸化如何促进细胞应激反应， 我们将讨论泛素磷酸化如何改变其相互作用特征以及与效应子的结合 途径。这项研究将为泛素和泛素生物学提供变革性的新见解 蛋白质稳态。第二个研究方向将解决人类葡萄糖转运蛋白如何被调节 泛素修饰和内吞运输。 GLUT家族的葡萄糖转运蛋白是关键的调节因子 细胞葡萄糖稳态，但对其运输的调节和质量控制仍然很差 特点。在这里，我们根据我们最近的发现描述了研究路线，即 GLUT1 内吞 贩运受到特定泛素修饰的调节。这些研究对细胞具有重要意义 葡萄糖稳态和人类疾病，包括 GLUT1 缺乏综合症和多种癌症。 总之，这些研究方向将导致对泛素代码、膜的更深入理解。 贩运和真核细胞的应激反应。