A RAD51–ADP double filament structure unveils the mechanism of filament dynamics in homologous recombination
Abstract ATP-dependent RAD51 recombinases play an essential role in eukaryotic homologous recombination by catalyzing a four-step process: 1) formation of a RAD51 single-filament assembly on ssDNA in the presence of ATP, 2) complementary DNA strand-exchange, 3) ATP hydrolysis transforming the RAD51...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-08-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-40672-5 |
| _version_ | 1797558696424767488 |
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| author | Shih-Chi Luo Min-Chi Yeh Yu-Hsiang Lien Hsin-Yi Yeh Huei-Lun Siao I-Ping Tu Peter Chi Meng-Chiao Ho |
| author_facet | Shih-Chi Luo Min-Chi Yeh Yu-Hsiang Lien Hsin-Yi Yeh Huei-Lun Siao I-Ping Tu Peter Chi Meng-Chiao Ho |
| author_sort | Shih-Chi Luo |
| collection | DOAJ |
| description | Abstract ATP-dependent RAD51 recombinases play an essential role in eukaryotic homologous recombination by catalyzing a four-step process: 1) formation of a RAD51 single-filament assembly on ssDNA in the presence of ATP, 2) complementary DNA strand-exchange, 3) ATP hydrolysis transforming the RAD51 filament into an ADP-bound disassembly-competent state, and 4) RAD51 disassembly to provide access for DNA repairing enzymes. Of these steps, filament dynamics between the ATP- and ADP-bound states, and the RAD51 disassembly mechanism, are poorly understood due to the lack of near-atomic-resolution information of the ADP-bound RAD51–DNA filament structure. We report the cryo-EM structure of ADP-bound RAD51–DNA filaments at 3.1 Å resolution, revealing a unique RAD51 double-filament that wraps around ssDNA. Structural analysis, supported by ATP-chase and time-resolved cryo-EM experiments, reveals a collapsing mechanism involving two four-protomer movements along ssDNA for mechanical transition between RAD51 single- and double-filament without RAD51 dissociation. This mechanism enables elastic change of RAD51 filament length during structural transitions between ATP- and ADP-states. |
| first_indexed | 2024-03-10T17:35:06Z |
| format | Article |
| id | doaj.art-fbde83d679064376b4c993bf26f916fa |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-10T17:35:06Z |
| publishDate | 2023-08-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-fbde83d679064376b4c993bf26f916fa2023-11-20T09:53:53ZengNature PortfolioNature Communications2041-17232023-08-0114111110.1038/s41467-023-40672-5A RAD51–ADP double filament structure unveils the mechanism of filament dynamics in homologous recombinationShih-Chi Luo0Min-Chi Yeh1Yu-Hsiang Lien2Hsin-Yi Yeh3Huei-Lun Siao4I-Ping Tu5Peter Chi6Meng-Chiao Ho7Institute of Biological Chemistry, Academia SinicaInstitute of Biological Chemistry, Academia SinicaInstitute of Statistical Science, Academia SinicaInstitute of Biochemical Sciences, National Taiwan UniversityInstitute of Statistical Science, Academia SinicaInstitute of Statistical Science, Academia SinicaInstitute of Biological Chemistry, Academia SinicaInstitute of Biological Chemistry, Academia SinicaAbstract ATP-dependent RAD51 recombinases play an essential role in eukaryotic homologous recombination by catalyzing a four-step process: 1) formation of a RAD51 single-filament assembly on ssDNA in the presence of ATP, 2) complementary DNA strand-exchange, 3) ATP hydrolysis transforming the RAD51 filament into an ADP-bound disassembly-competent state, and 4) RAD51 disassembly to provide access for DNA repairing enzymes. Of these steps, filament dynamics between the ATP- and ADP-bound states, and the RAD51 disassembly mechanism, are poorly understood due to the lack of near-atomic-resolution information of the ADP-bound RAD51–DNA filament structure. We report the cryo-EM structure of ADP-bound RAD51–DNA filaments at 3.1 Å resolution, revealing a unique RAD51 double-filament that wraps around ssDNA. Structural analysis, supported by ATP-chase and time-resolved cryo-EM experiments, reveals a collapsing mechanism involving two four-protomer movements along ssDNA for mechanical transition between RAD51 single- and double-filament without RAD51 dissociation. This mechanism enables elastic change of RAD51 filament length during structural transitions between ATP- and ADP-states.https://doi.org/10.1038/s41467-023-40672-5 |
| spellingShingle | Shih-Chi Luo Min-Chi Yeh Yu-Hsiang Lien Hsin-Yi Yeh Huei-Lun Siao I-Ping Tu Peter Chi Meng-Chiao Ho A RAD51–ADP double filament structure unveils the mechanism of filament dynamics in homologous recombination Nature Communications |
| title | A RAD51–ADP double filament structure unveils the mechanism of filament dynamics in homologous recombination |
| title_full | A RAD51–ADP double filament structure unveils the mechanism of filament dynamics in homologous recombination |
| title_fullStr | A RAD51–ADP double filament structure unveils the mechanism of filament dynamics in homologous recombination |
| title_full_unstemmed | A RAD51–ADP double filament structure unveils the mechanism of filament dynamics in homologous recombination |
| title_short | A RAD51–ADP double filament structure unveils the mechanism of filament dynamics in homologous recombination |
| title_sort | rad51 adp double filament structure unveils the mechanism of filament dynamics in homologous recombination |
| url | https://doi.org/10.1038/s41467-023-40672-5 |
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