Spatio-Temporal Organisation of Genome Surveillance in Live Cells

基因组监测对细胞功能、癌症避免和发育的许多方面都至关重要，它由一系列DNA修复和损伤反应途径组成。损害监测的缺陷会导致严重的遗传疾病。人们对这些通路的机制有不同程度的详细了解，本研究的目的是了解不同DNA损伤处理前后细胞核内蛋白质组分的动态变化，以及不同通路之间的相互关系。用GFP及其光谱变体标记的正常和特征性突变蛋白，要么可以从参与的实验室获得，要么将作为提案的一部分产生。用光漂白结合全细胞或用紫外光或电离辐射局部照射后荧光恢复的变化来测量蛋白质的迁移率。合作伙伴的互补性来自于他们在以下方面的专长：(1)不同的监测途径和提供标记蛋白质；(2)先进的显微技术；(3)提供不同类型的局部照射；(4)计算机模拟。通过整合参与小组的不同专业知识、独特的材料和试剂以及专门的设备，该提案形成了一种全面的、多学科的方法来了解哺乳动物细胞基因组监测的动态。

Surveillance of the genome, which is vital for cellular function, cancer avoidance and many aspects of development, is comprised of a series of DNA repair and damage response pathways. Defects in damage surveillance result in severe genetic disorders. The mechanisms of these pathways are understood in varying degrees of detail, and the aim of this proposal is to understand the dynamics of the protein constituents within the cell nucleus before and after different DNA damaging treatments, as well as the inter-relationships between the different pathways. Normal and characterised mutant proteins tagged with GFP and its spectral variants, are either available from the participating laboratories or will be generated as part of the proposal. Motilities of the proteins are measured using variations of fluorescence recovery after photobleaching combined with whole cell or localised irradiation with either UV light or ionizing radiation. The complementarity of the partners comes from their expertise in (1) different surveillance pathways and provision of tagged proteins; (2) advanced microscopic techniques; (3) delivery of different types of localised irradiation; (4) computer simulation. Through the integration of the different expertises, unique materials and reagents, and specialized equipment from the participating groups, the proposal forms a comprehensive and multidisciplinary approach to understanding the dynamics of genome surveillance in mammalian cells.