From single-molecule to synchrotron: New tools for investigating the molecular mechanisms of Aicardi-Goutires Syndrome and other aberrant DNA diseases

从单分子到同步加速器：研究 Aicardi-Goutires 综合征和其他异常 DNA 疾病分子机制的新工具

• 批准号: 2289215
• 金额: --
• 依托单位: University of Sheffield
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2289215
• 关键词: single; molecule; synchrotron; New; tools

This studentship is focused on developing new biophysical tools to understand the molecular basis of aberrant DNA diseases. You will be part of a new, multidisciplinary collaboration between the Sheffield Institute for Nucleic Acids (http://genome.sheffield.ac.uk/) at the University of Sheffield, and Diamond Light Source Ltd (https://www.diamond.ac.uk/Home.html). As such it is likely the project will attract an enhanced stipend (as an iCASE award).A major challenge to genome stability is the presence of small amounts of RNA interspersed within DNA. Recent studies report the levels of ribose incorporation in mammalian genomes to be >1 million nucleotides per day, making it the most frequent source of cellular DNA damage in eukaryotes. This aberrant RNA must be removed from the DNA. The human enzymes responsible for this are RNAseH1 - which recognises runs of at least four ribonucleotides), and RNAseH2 - which can detect a single ribonucleotide, hydrolysing the 5'-phosphodiester bond leading to its removal.RNaseH2 is formed from three polypeptides; RNAse H2A, H2B, and H2C. Single mutations in all these subunits have been found in patients suffering from Aicardi-Goutières Syndrome, an inflammatory disease effecting the brain and skin. Additionally, we recently reported that accumulation of DNA/RNA hybrids cause neurodegeneration in motor neuron disease and frontotemporal dementia (1). The goal of this project is to understand how these crucial enzymes locate ribonucleotides (one additional oxygen atom) within a vast sea of normal DNA and the effects of disease-causing mutations on this process.You will use established single-molecule FRET techniques in the Craggs Lab (2), alongside new Small Angle X-ray Scattering methods (using gold nano-particles) that you will develop in collaboration with beamline scientists at Diamond, both in combination with computational modelling (with our collaborator in Denmark, Prof Kresten Lindorff-Larsen https://www1.bio.ku.dk/english/research/bms/research/sbinlab/groups/kll/), to characterize the conformational dynamics of aberrant nucleic acids in solution, both in the presence and absence of RNaseH proteins.This studentship provides a unique opportunity to be trained across a range of cutting-edge biophysical techniques and apply them to a fundamental question on the molecular understanding of disease. We are looking for creative individuals capable of working across disciplines.

这个学生奖学金的重点是开发新的生物物理工具，以了解异常DNA疾病的分子基础。您将成为谢菲尔德大学谢菲尔德核酸研究所（http：//genome.sheffield.ac.uk/）和Diamond Light Source Ltd（https：//www.diamond.ac.uk/Home.html）之间新的多学科合作的一部分。因此，该项目很可能会吸引更多的津贴（作为iCASE奖）。基因组稳定性的一个主要挑战是DNA中存在少量的RNA。最近的研究报道，哺乳动物基因组中核糖掺入的水平为每天>1百万个核苷酸，使其成为真核生物中细胞DNA损伤的最常见来源。这种异常的RNA必须从DNA中去除。负责此的人类酶是RNAseH 1-其识别至少四个核糖核苷酸的运行）和RNAseH 2-其可以检测单个核糖核苷酸，水解5 '-磷酸二酯键导致其去除。RNaseH 2由三种多肽形成; RNAse H2 A，H2 B和H2 C。所有这些亚基中的单一突变已在患有Aicardi-Goutières综合征的患者中发现，Aicardi-Goutières综合征是一种影响大脑和皮肤的炎性疾病。此外，我们最近报道了DNA/RNA杂合体的积累导致运动神经元疾病和额颞叶痴呆的神经变性（1）。这个项目的目标是了解这些关键酶如何定位核糖核苷酸（一个额外的氧原子）在一个巨大的海洋正常的DNA和致病突变对这一过程的影响。你将使用建立单分子FRET技术在克雷格斯实验室（2），以及新的小角X射线散射方法（使用金纳米粒子），您将与Diamond的光束线科学家合作开发，（与我们在丹麦的合作者Kresten Lindorff-Larsen教授https：//www1.bio.ku.dk/english/research/bms/research/sbinlab/groups/kll/），以表征溶液中异常核酸的构象动力学，在RNaseH蛋白存在和不存在的情况下。这个学生提供了一个独特的机会，在一系列尖端的生物物理技术的培训，并将其应用到疾病的分子理解的基本问题。我们正在寻找能够跨学科工作的创造性人才。