Newton001: Targeting the surface proteome of Trypanosoma cruzi

原生动物寄生虫的表面是与宿主的界面，在入侵、免疫逃避、信号传递和环境感知等方面发挥着重要作用。我们最近在锥虫体内发现了一个保守的泛素化途径，它控制着重要的跨膜结构域蛋白的表面拷贝数，这为合理地控制这些寄生虫的跨膜结构域蛋白丰度提供了一种手段，并首次评估了它们对宿主细胞入侵和毒力的重要性。本应用旨在评估克氏锥虫表面蛋白泛素化途径的意义：1。2.寄主细胞侵染与寄主生命周期的关系。2 .表面涂层的维持，特别是跨膜结构域蛋白的拷贝数量及其对整体表面蛋白质组的影响。通过体外生存能力评估，作为治疗靶点的可能价值。这项工作将结合使用CRISPR/Cas9方法的最先进的基因组编辑技术，以及SILAC蛋白质组学对表面/细胞蛋白质组进行无偏分析。

The surface of protozoan parasites represent the interface with the host, with many roles in invasion, immune evasion, signalling, and environmental sensing. We recently identified a conserved ubiquitylation pathway in trypanosomes that controls the surface copy number of important trans-membrane domain proteins, and which provides a means for the first time to manipulate trans-membrane domain protein abundance in these parasites in a rational manor, and to assess their importance to host cell invasion and virulence. This application seeks to assess the significance of the surface protein ubiquitylation pathway in Trypanosoma cruzi for: 1. Parasite viability, host cell invasion and life cycle progression,2. Maintenance of the surface coat, specificallycopy number of trans-membrane domain proteins and their impact on the global surface proteome, 3. Possible value as a therapeutic target, by assessment of viability in vitro.The work will combine state of the art genome editing using CRISPR/Cas9 approaches, together with SILAC proteomics for unbiased analysis of the surface/cellular proteome.