Structural and functional insights into the interaction between Ku70/80 and Pol X family polymerases in NHEJ

IF 15.7 1区综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES

Nature Communications Pub Date : 2025-05-06 DOI:10.1038/s41467-025-59133-2

Philippe Frit, Himani Amin, Sayma Zahid, Nadia Barboule, Chloe Hall, Gurdip Matharu, Steven W. Hardwick, Jeanne Chauvat, Sébastien Britton, Dima Y. Chirgadze, Virginie Ropars, Jean-Baptiste Charbonnier, Patrick Calsou, Amanda K. Chaplin

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Abstract

Non-homologous end joining (NHEJ) is the main repair pathway for double-strand DNA breaks (DSBs) in mammals. DNA polymerases lambda (Pol λ) and mu (Pol μ), members of the Pol X family, play a key role in this process. However, their interaction within the NHEJ complexes is unclear. Here, we present cryo-EM structures of Pol λ in complex with the DNA-PK long-range synaptic complex, and Pol μ bound to Ku70/80-DNA. These structures identify interaction sites between Ku70/80 and Pol X BRCT domains. Using mutants at the proteins interface in functional assays including cell transfection with an original gap-filling reporter, we define the role of the BRCT domain in the recruitment and activity of the two Pol X members in NHEJ and in their contribution to cell survival following DSBs. Finally, we propose a unified model for the interaction of all Pol X members with Ku70/80.

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NHEJ中Ku70/80与Pol X家族聚合酶相互作用的结构和功能研究

非同源末端连接（Non-homologous end joining， NHEJ）是哺乳动物DNA双链断裂（DSBs）的主要修复途径。DNA聚合酶lambda （Pol λ）和mu （Pol μ）是Pol X家族的成员，在这一过程中起着关键作用。然而，它们在NHEJ复合物中的相互作用尚不清楚。本文研究了Pol λ与DNA-PK远程突触复合物的低温电镜结构，以及Pol μ与Ku70/80-DNA的结合。这些结构确定了Ku70/80和Pol X BRCT结构域之间的相互作用位点。在功能分析中使用蛋白质界面的突变体，包括用原始缺口填充报告细胞转染，我们定义了BRCT结构域在NHEJ中两个Pol X成员的招募和活性以及它们对DSBs后细胞存活的贡献中的作用。最后，我们提出了所有Pol X成员与Ku70/80相互作用的统一模型。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Nature Communications Biological Science Disciplines-

CiteScore

24.90

自引率

2.40%

发文量

6928

审稿时长

3.7 months

期刊介绍： Nature Communications, an open-access journal, publishes high-quality research spanning all areas of the natural sciences. Papers featured in the journal showcase significant advances relevant to specialists in each respective field. With a 2-year impact factor of 16.6 (2022) and a median time of 8 days from submission to the first editorial decision, Nature Communications is committed to rapid dissemination of research findings. As a multidisciplinary journal, it welcomes contributions from biological, health, physical, chemical, Earth, social, mathematical, applied, and engineering sciences, aiming to highlight important breakthroughs within each domain.