DNA DAMAGE REPAIR

DNA损伤修复

• 批准号: 8169127
• 负责人: Hironori Funabiki
• 金额: $ 0.7万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169127
• 关键词: Cells; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA; Double Strand Break Repair; Excision; F Box Domain; Funding; G22P1 gene; Grant; Institution; Length; Link; Manuscripts; Mediating; Methods; Oligonucleotides; Pathway interactions; Process; Proteins; Publishing; Recruitment Activity; Research; Research Personnel; Resources; Source; Ubiquitin; United States National Institutes of Health; Work; XRCC5 gene; Xenopus laevis; egg; member; repaired; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Ku70/Ku80 heterodimer, or Ku, is the central component of the nonhomologous end joining (NHEJ) pathway of double strand break (DSB) repair. Because Ku forms a ring through which the DSB threads, it likely becomes topologically attached to DNA during repair. The mechanism for its removal was unknown. Using a method to identify proteins recruited to DSBs in Xenopus laevis egg extract, we show that DSB-containing DNAs accumulate members of the Skp1-Cul1-F-box complex and K48-linked polyubiquitylated proteins in addition to known repair proteins. We demonstrate that Ku80 is degraded in response to DSBs in a ubiquitin-mediated manner. Strikingly, K48-linked polyubiquitylation, but not proteasomal degradation, is required for the efficient removal of Ku80 from DNA. This removal is DNA length dependent, as Ku80 is retained on duplex oligonucleotides. Finally, NHEJ completion and removal of Ku80 from DNA are independent from one another. We propose that DSB-induced ubiquitylation of Ku80 provides a mechanism to efficiently eliminate Ku from DNA for pre- and postrepair processes. A manuscript describing this work has been published: Ku80 removal from DNA through double strand break-induced ubiquitylation. Postow L, Ghenoiu C, Woo EM, Krutchinsky AN, Chait BT, Funabiki H. J Cell Biol. 2008 11;182(3):467-79.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 KU70/KU80异二聚体（KU）是双链断裂（DSB）修复的非同源末端连接（NHEJ）途径的核心组成部分。由于KU形成了DSB螺纹的环，因此在修复过程中可能会在DNA上拓扑结合。其去除的机制尚不清楚。使用一种方法来鉴定在Xenopus laevis Egg提取物中募集到DSB的蛋白质，我们表明，含DSB的DNA会累积SKP1-CUL1-F-box复合物的成员，除了已知的修复蛋白外，除了已知的蛋白质外，还积累了K48连接的多偶联性蛋白。我们证明，Ku80以泛素介导的方式响应DSB而降解。引人注目的是，有效从DNA中有效去除KU80所必需的K48连接的多泛素化，但不是蛋白酶体降解。该去除取决于DNA的长度，因为KU80保留在双链寡核苷酸上。最后，NHEJ完成和从DNA中删除KU80是独立的。我们建议DSB诱导的KU80的泛素化提供了一种机制，可以有效地从DNA中消除ku进行前和后epperpair过程。描述这项工作的手稿已经发表： 从DNA通过双链断裂诱导的泛素化去除KU80。 Postow L，Ghenoiu C，Woo EM，Kr​​utchinsky AN，Chait BT，Funabiki H. J Cell Biol。 2008 11; 182（3）：467-79。