Function of the UBZ-containing ATPase Wrnip1 in DNA replication and damage response

含有 UBZ 的 ATPase Wrnip1 在 DNA 复制和损伤反应中的功能

• 批准号: 72189856
• 负责人: Professor Dr. Ivan Dikic
• 金额: --
• 依托单位: Institut für Biochemie II
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2008
• 资助国家: 德国
• 起止时间: 2007-12-31 至 2013-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/72189856?language=en
• 关键词: Function; UBZ; containing; ATPase; Wrnip1

Ubiquitin (Ub) and Ubiquitin-like modifiers (Ubls) are among the most diverse moieties covalentiy attached to proteins to maintain homeostasis under normal conditions and during stress such as DNA damage. We have previously shown that multiple proteins involved in the DNA damage response (DDR) - including the Y-family polymerases kappa and eta, Werner helicase (Wrn)-interacting protein 1 (Wmip1), FAN-1 nuclease and Rad18 ubiquitin ligase - contain Ub-binding Zn finger (UBZ) domains that regulate their sub-nuclear localization inside focal structures. Here we propose to study the role of Wrnip1 in DNA replication, telomere maintenance and genome stability. We have found that Wmip1 foci co-localize with heterochromatic regions including centromeres and telomeres and that Wrnip1 interacts with the telomeric protein TRF2 (telomere repeat factor 2) but not TRF1. We have also found that acute Wrnip1 depletion by RNA interference causes chromosomal instability involving both centromeres and telomeres, To dissect the functional role of Wrnip1, we will combine biochemical and cellular methods to study: a) the composition of Wrnip1-associated complexes; b) the dynamics of Wrnip1 foci in living cells under replication stress; c) the relevance of Wrnip1 for telomere maintenance, DNA replication and genome stability; d) the effects of the deletion of WRNIP1 and associated genes such as RAD18 and WRN in defined tissues and organs in genetically-engineered mice models. We expect that these studies will shed light onto the physiological role and mechanism of action of Wrnip1 during DDR.

泛素(Ub)和泛素样修饰物(UBL)是蛋白质分子中种类最多的共价基团，在正常条件下和在DNA损伤等应激状态下维持体内平衡。我们已经证明，参与DNA损伤反应(DDR)的多种蛋白质--包括Y-家族聚合酶Kappa和Eta、Werner Helicase(WRN)相互作用蛋白1(Wmip1)、FAN-1核酸酶和Rad18泛素连接酶--含有Ub结合的锌指(UBZ)结构域，调节其在焦点结构中的亚核定位。在这里，我们打算研究Wrnip1在DNA复制、端粒维持和基因组稳定性中的作用。我们发现Wmip1与着丝粒和端粒等异染色区共定位，Wrnip1与端粒蛋白TRF2(端粒重复因子2)相互作用，但不与TRF1相互作用。我们还发现，通过RNA干扰导致的Wrnip1急性缺失会导致涉及着丝粒和端粒的染色体不稳定，为了剖析Wrnip1的功能作用，我们将结合生化和细胞方法来研究：a)Wrnip1相关复合体的组成；b)复制应激下活细胞中Wrnip1焦点的动态；c)Wrnip1与端粒维持、DNA复制和基因组稳定性的相关性；d)在基因工程小鼠的特定组织和器官中，WRNIP1和相关基因如RAD18和WRN的缺失所产生的影响。我们期望这些研究将有助于阐明Wrnip1在DDR中的生理作用和作用机制。