Structural and functional insights into the interaction between Ku70/80 and Pol X family polymerases in NHEJ
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...
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Nature Portfolio
2025-05-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-59133-2 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Philippe Frit |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-28cd83edd27a4b9abaebbc20f42c1f3f |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-28cd83edd27a4b9abaebbc20f42c1f3f2025-08-20T03:53:13ZengNature PortfolioNature Communications2041-17232025-05-0116111410.1038/s41467-025-59133-2Structural and functional insights into the interaction between Ku70/80 and Pol X family polymerases in NHEJPhilippe Frit0Himani Amin1Sayma Zahid2Nadia Barboule3Chloe Hall4Gurdip Matharu5Steven W. Hardwick6Jeanne Chauvat7Sébastien Britton8Dima Y. Chirgadze9Virginie Ropars10Jean-Baptiste Charbonnier11Patrick Calsou12Amanda K. Chaplin13Institut de Pharmacologie et Biologie Structurale (IPBS), Université de Toulouse, CNRS, Université Toulouse III—Paul Sabatier (UT3)Leicester Institute for Structural and Chemical Biology, Department of Molecular and Cell Biology, University of LeicesterLeicester Institute for Structural and Chemical Biology, Department of Molecular and Cell Biology, University of LeicesterInstitut de Pharmacologie et Biologie Structurale (IPBS), Université de Toulouse, CNRS, Université Toulouse III—Paul Sabatier (UT3)Leicester Institute for Structural and Chemical Biology, Department of Molecular and Cell Biology, University of LeicesterLeicester Institute for Structural and Chemical Biology, Department of Molecular and Cell Biology, University of LeicesterDepartment of Biochemistry, Sanger Building, University of CambridgeInstitut de Pharmacologie et Biologie Structurale (IPBS), Université de Toulouse, CNRS, Université Toulouse III—Paul Sabatier (UT3)Institut de Pharmacologie et Biologie Structurale (IPBS), Université de Toulouse, CNRS, Université Toulouse III—Paul Sabatier (UT3)Department of Biochemistry, Sanger Building, University of CambridgeInstitute for Integrative Biology of the Cell (I2BC), Institute Joliot, CEA, CNRS, Université Paris-SaclayInstitute for Integrative Biology of the Cell (I2BC), Institute Joliot, CEA, CNRS, Université Paris-SaclayInstitut de Pharmacologie et Biologie Structurale (IPBS), Université de Toulouse, CNRS, Université Toulouse III—Paul Sabatier (UT3)Leicester Institute for Structural and Chemical Biology, Department of Molecular and Cell Biology, University of LeicesterAbstract 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.https://doi.org/10.1038/s41467-025-59133-2 |
| spellingShingle | 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 Structural and functional insights into the interaction between Ku70/80 and Pol X family polymerases in NHEJ Nature Communications |
| title | Structural and functional insights into the interaction between Ku70/80 and Pol X family polymerases in NHEJ |
| title_full | Structural and functional insights into the interaction between Ku70/80 and Pol X family polymerases in NHEJ |
| title_fullStr | Structural and functional insights into the interaction between Ku70/80 and Pol X family polymerases in NHEJ |
| title_full_unstemmed | Structural and functional insights into the interaction between Ku70/80 and Pol X family polymerases in NHEJ |
| title_short | Structural and functional insights into the interaction between Ku70/80 and Pol X family polymerases in NHEJ |
| title_sort | structural and functional insights into the interaction between ku70 80 and pol x family polymerases in nhej |
| url | https://doi.org/10.1038/s41467-025-59133-2 |
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