Mechanisms of deubiquitination in DNA repair

DNA 修复中的去泛素化机制

• 批准号: 2881072
• 金额: --
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2881072
• 关键词: Mechanisms; deubiquitination; DNA; repair

The covalent attachment of ubiquitin (Ub) to substrates is a post-translational modification (PTM) that has wide-ranging functions in determining protein fate. As such, the Ub system has a role in myriad cellular pathways, including proteostasis, inflammation, and the DNA damage response. The maturation of Ub precursor proteins and the removal of Ub and polyUb chains from substrates is carried out by dedicated proteases called deubiquitinases (DUBs), which regulate the amplitude and duration of Ub signaling in cells. There are approximately 100 DUBs encoded in the human genome that can be classified into seven distinct families based on their structural features and evolutionary ancestry. Zinc finger-containing Ub peptidase 1 (ZUP1) was recently discovered as the founding, and only member, of a seventh DUB class that cleaves long K63-linked polyUb chains, with a putative role in the DNA damage response. However, despite these exciting recent findings there are several mechanistic aspects of ZUP1 function that remain unknown, including how it specifically hydrolyses K63-linked polyUb versus all other Ub linkage types, how it recognizes long K63-linked polyUb chains, and how it is activated by its binding partner, the RPA complex.To address these unresolved aspects of this unique DUB class, there are three main aims for this DPhil project that, if accomplished, will significantly deepen our understanding of ZUP1 and DUB function in general.1. Determine the mechanism for ZUP1 K63-linkage specificity, using structural biology, biochemistry, and cell biology.2. Determine the mechanism of polyUb chain recognition by ZUP1 using single molecule imaging and biophysical approaches.3. Determine the mechanism of RPA-dependent stimulation of ZUP1 activity using the approaches listed in #1 and #2.Through these aims, the candidate will obtain training and development in a range of integrated cutting-edge and quantitative approaches that will enable them to take their research beyond the state-of-the-art. Coupled with additional skills training and development opportunities, this project will provide an excellent foundation and springboard for the candidate's future career.

泛素（Ub）与底物的共价附着是一种翻译后修饰（PTM），在决定蛋白质命运方面具有广泛的功能。因此，Ub系统在多种细胞途径中发挥作用，包括蛋白质停滞、炎症和DNA损伤反应。Ub前体蛋白的成熟以及Ub和polyb链从底物上的去除是由称为去泛素酶（DUBs）的专用蛋白酶进行的，该蛋白酶调节细胞中Ub信号传导的幅度和持续时间。人类基因组中编码了大约100个dub，根据它们的结构特征和进化祖先，可以将它们分为7个不同的家族。含有锌指的Ub肽酶1 （ZUP1）最近被发现是第7类DUB的唯一成员，它可以切割长k63连接的polyb链，并在DNA损伤反应中发挥作用。然而，尽管最近有这些令人兴奋的发现，ZUP1功能的几个机制方面仍然未知，包括它如何特异性地水解k63连接的polyb而不是所有其他Ub连接类型，它如何识别长k63连接的polyb链，以及它如何被其结合伙伴RPA复合物激活。为了解决这个独特的DUB类的这些未解决的方面，这个DPhil项目有三个主要目标，如果完成，将大大加深我们对ZUP1和DUB功能的理解。利用结构生物学、生物化学和细胞生物学，确定ZUP1 k63连锁特异性的机制。利用单分子成像和生物物理方法确定ZUP1对polyb链识别的机制。使用#1和#2中列出的方法确定rpa依赖性刺激ZUP1活性的机制。通过这些目标，候选人将获得一系列综合前沿和定量方法的培训和发展，这将使他们的研究超越最先进的水平。加上额外的技能培训和发展机会，该项目将为候选人未来的职业生涯提供良好的基础和跳板。