同源重组修复过程中 H2B 单泛素化在 D 环代谢中的多方面作用

Shih-Hsun Hung, Yuan Liang, Wolf Dietrich Heyer
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引用次数: 0

摘要

DNA 双链断裂的修复对于维持基因组的完整性至关重要,这主要是通过同源重组(HR)来实现的。核小体由包裹在组蛋白八聚体周围的 DNA 组成,为启动 HR 的末端重组提供了天然屏障,但其对同源搜索、DNA 链侵入和修复合成等下游 HR 步骤的影响仍有待确定。位移环(D-loop)在HR中起着关键作用,但染色质动力学对D-loop代谢的影响仍不清楚。我们利用物理 D-loop 捕获(DLC)和 D-loop 延伸(DLE)试验追踪 HR 中间体,并通过遗传分析揭示了 H2B 单泛素化(H2Bubi)影响 HR 修复过程中的多个步骤。我们推断,H2Bubi 调节染色质结构,不仅促进组蛋白降解以形成新生 D 环,还通过核小体组装稳定扩展的 D 环。此外,H2Bubi 还通过 Rad9 招募调节 DNA 切除,抑制反馈控制机制,从而抑制 D 环在超切除末端的形成和延伸。通过物理和遗传实验来确定修复结果,我们证明了 H2Bubi 在防止断裂诱导复制从而促进基因组稳定性方面起着至关重要的作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Multifaceted roles of H2B mono-ubiquitylation in D-loop metabolism during homologous recombination repair
Repairing DNA double-strand breaks is crucial for maintaining genome integrity, which occurs primarily through homologous recombination (HR) in S. cerevisiae. Nucleosomes, composed of DNA wrapped around a histone octamer, present a natural barrier to end-resection to initiate HR, but the impact on the downstream HR steps of homology search, DNA strand invasion and repair synthesis remain to be determined. Displacement loops (D-loops) play a pivotal role in HR, yet the influence of chromatin dynamics on D-loop metabolism remains unclear. Using the physical D-loop capture (DLC) and D-loop extension (DLE) assays to track HR intermediates, we employed genetic analysis to reveal that H2B mono-ubiquitylation (H2Bubi) affects multiple steps during HR repair. We infer that H2Bubi modulates chromatin structure, not only promoting histone degradation for nascent D-loop formation but also stabilizing extended D-loops through nucleosome assembly. Furthermore, H2Bubi regulates DNA resection via Rad9 recruitment to suppress a feedback control mechanism that dampens D-loop formation and extension at hyper-resected ends. Through physical and genetic assays to determine repair outcomes, we demonstrate that H2Bubi plays a crucial role in preventing break-induced replication and thus promoting genomic stability.
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