Mechanism of Rad51 filament formation by Rad52 and Rad55-Rad57 in homologous recombination
Abstract Homologous recombination (HR) repairs double-stranded DNA breaks (DSBs) by generating single-stranded DNA (ssDNA), which is initially coated by Replication Protein A (Rpa). Rad51, a recombinase, catalyzes strand invasion but binds ssDNA with lower affinity than Rpa, necessitating mediator p...
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Nature Portfolio
2025-07-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-61811-0 |
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| author | Jaigeeth Deveryshetty Ayush Mistry Sushil Pangeni Mohamed Ghoneim Monica Tokmina-Lukaszewska Steven K. Gore Jie Liu Vikas Kaushik Simrithaa Karunakaran Angela Taddei Wolf-Dietrich Heyer Taekjip Ha Brian Bothner Edwin Antony |
| author_facet | Jaigeeth Deveryshetty Ayush Mistry Sushil Pangeni Mohamed Ghoneim Monica Tokmina-Lukaszewska Steven K. Gore Jie Liu Vikas Kaushik Simrithaa Karunakaran Angela Taddei Wolf-Dietrich Heyer Taekjip Ha Brian Bothner Edwin Antony |
| author_sort | Jaigeeth Deveryshetty |
| collection | DOAJ |
| description | Abstract Homologous recombination (HR) repairs double-stranded DNA breaks (DSBs) by generating single-stranded DNA (ssDNA), which is initially coated by Replication Protein A (Rpa). Rad51, a recombinase, catalyzes strand invasion but binds ssDNA with lower affinity than Rpa, necessitating mediator proteins like Rad52 (yeast) or BRCA2 (humans) for Rad51 loading. The mechanisms of this exchange remain unclear. We show that Saccharomyces cerevisiae Rad52 uses its disordered C-terminus to sort polydisperse Rad51 into discrete monomers. Using fluorescent-Rad51 and single-molecule optical tweezers, we visualize Rad52-mediated Rad51 filament formation on Rpa-coated ssDNA, preferentially at ssDNA–dsDNA junctions. Deleting the C-terminus of Rad52 disrupts Rad51 sorting and loading. Addition of the Rad51 paralog Rad55–Rad57 enhances Rad51 binding by ~60%. Despite structural differences, Rad52 and BRCA2 share conserved functional features. We propose a unified “Sort, Stack & Extend” (SSE) mechanism by which mediator proteins and paralogs coordinate Rad51 filament assembly during HR. |
| format | Article |
| id | doaj-art-50df6a168b37484a928ee4d2bcf0a41b |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-50df6a168b37484a928ee4d2bcf0a41b2025-08-20T03:46:28ZengNature PortfolioNature Communications2041-17232025-07-0116111710.1038/s41467-025-61811-0Mechanism of Rad51 filament formation by Rad52 and Rad55-Rad57 in homologous recombinationJaigeeth Deveryshetty0Ayush Mistry1Sushil Pangeni2Mohamed Ghoneim3Monica Tokmina-Lukaszewska4Steven K. Gore5Jie Liu6Vikas Kaushik7Simrithaa Karunakaran8Angela Taddei9Wolf-Dietrich Heyer10Taekjip Ha11Brian Bothner12Edwin Antony13Department of Biochemistry and Molecular Biology, St. Louis University School of MedicineDepartment of Biochemistry and Molecular Biology, St. Louis University School of MedicineDepartment of Biophysics, Johns Hopkins UniversityDepartment of Biochemistry and Molecular Biology, St. Louis University School of MedicineDepartment of Chemistry and Biochemistry, Montana State UniversityDepartment of Microbiology and Molecular Genetics, University of CaliforniaDepartment of Microbiology and Molecular Genetics, University of CaliforniaDepartment of Biochemistry and Molecular Biology, St. Louis University School of MedicineDepartment of Biochemistry and Molecular Biology, St. Louis University School of MedicineInstitut Curie, Université PSL, Sorbonne University, CNRS, Nuclear DynamicsDepartment of Microbiology and Molecular Genetics, University of CaliforniaProgram in Cellular and Molecular Medicine, Children’s HospitalDepartment of Chemistry and Biochemistry, Montana State UniversityDepartment of Biochemistry and Molecular Biology, St. Louis University School of MedicineAbstract Homologous recombination (HR) repairs double-stranded DNA breaks (DSBs) by generating single-stranded DNA (ssDNA), which is initially coated by Replication Protein A (Rpa). Rad51, a recombinase, catalyzes strand invasion but binds ssDNA with lower affinity than Rpa, necessitating mediator proteins like Rad52 (yeast) or BRCA2 (humans) for Rad51 loading. The mechanisms of this exchange remain unclear. We show that Saccharomyces cerevisiae Rad52 uses its disordered C-terminus to sort polydisperse Rad51 into discrete monomers. Using fluorescent-Rad51 and single-molecule optical tweezers, we visualize Rad52-mediated Rad51 filament formation on Rpa-coated ssDNA, preferentially at ssDNA–dsDNA junctions. Deleting the C-terminus of Rad52 disrupts Rad51 sorting and loading. Addition of the Rad51 paralog Rad55–Rad57 enhances Rad51 binding by ~60%. Despite structural differences, Rad52 and BRCA2 share conserved functional features. We propose a unified “Sort, Stack & Extend” (SSE) mechanism by which mediator proteins and paralogs coordinate Rad51 filament assembly during HR.https://doi.org/10.1038/s41467-025-61811-0 |
| spellingShingle | Jaigeeth Deveryshetty Ayush Mistry Sushil Pangeni Mohamed Ghoneim Monica Tokmina-Lukaszewska Steven K. Gore Jie Liu Vikas Kaushik Simrithaa Karunakaran Angela Taddei Wolf-Dietrich Heyer Taekjip Ha Brian Bothner Edwin Antony Mechanism of Rad51 filament formation by Rad52 and Rad55-Rad57 in homologous recombination Nature Communications |
| title | Mechanism of Rad51 filament formation by Rad52 and Rad55-Rad57 in homologous recombination |
| title_full | Mechanism of Rad51 filament formation by Rad52 and Rad55-Rad57 in homologous recombination |
| title_fullStr | Mechanism of Rad51 filament formation by Rad52 and Rad55-Rad57 in homologous recombination |
| title_full_unstemmed | Mechanism of Rad51 filament formation by Rad52 and Rad55-Rad57 in homologous recombination |
| title_short | Mechanism of Rad51 filament formation by Rad52 and Rad55-Rad57 in homologous recombination |
| title_sort | mechanism of rad51 filament formation by rad52 and rad55 rad57 in homologous recombination |
| url | https://doi.org/10.1038/s41467-025-61811-0 |
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