Rad52 Oligomeric N-Terminal Domain Stabilizes Rad51 Nucleoprotein Filaments and Contributes to Their Protection against Srs2
Homologous recombination (HR) depends on the formation of a nucleoprotein filament of the recombinase Rad51 to scan the genome and invade the homologous sequence used as a template for DNA repair synthesis. Therefore, HR is highly accurate and crucial for genome stability. Rad51 filament formation i...
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doaj-80f55cb50bae4836b609b16118aa67732021-06-30T23:54:52ZengMDPI AGCells2073-44092021-06-01101467146710.3390/cells10061467Rad52 Oligomeric N-Terminal Domain Stabilizes Rad51 Nucleoprotein Filaments and Contributes to Their Protection against Srs2Emilie Ma0Laurent Maloisel1Léa Le Falher2Raphaël Guérois3Eric Coïc4Université de Paris and Université Paris-Saclay, Inserm, LGRM/iRCM/IBFJ-CEA, UMR Stabilité Génétique Cellules Souches et Radiations, F-92265 Fontenay-Aux-Roses, FranceUniversité de Paris and Université Paris-Saclay, Inserm, LGRM/iRCM/IBFJ-CEA, UMR Stabilité Génétique Cellules Souches et Radiations, F-92265 Fontenay-Aux-Roses, FrancePresent address: Precision Oncology Genomics, Oncology Therapeutic Area, Sanofi R&D, F-94403 Vitry-Sur-Seine, FranceUniversité Paris Saclay, CNRS, LBSR/i2BC-CEA, Institute for Integrative Biology of the Cell (I2BC), F-91198 Gif-Sur-Yvette, FranceUniversité de Paris and Université Paris-Saclay, Inserm, LGRM/iRCM/IBFJ-CEA, UMR Stabilité Génétique Cellules Souches et Radiations, F-92265 Fontenay-Aux-Roses, FranceHomologous recombination (HR) depends on the formation of a nucleoprotein filament of the recombinase Rad51 to scan the genome and invade the homologous sequence used as a template for DNA repair synthesis. Therefore, HR is highly accurate and crucial for genome stability. Rad51 filament formation is controlled by positive and negative factors. In <i>Saccharomyces cerevisiae,</i> the mediator protein Rad52 catalyzes Rad51 filament formation and stabilizes them, mostly by counteracting the disruptive activity of the translocase Srs2. Srs2 activity is essential to avoid the formation of toxic Rad51 filaments, as revealed by Srs2-deficient cells. We previously reported that Rad52 SUMOylation or mutations disrupting the Rad52–Rad51 interaction suppress Rad51 filament toxicity because they disengage Rad52 from Rad51 filaments and reduce their stability. Here, we found that mutations in Rad52 N-terminal domain also suppress the DNA damage sensitivity of Srs2-deficient cells. Structural studies showed that these mutations affect the Rad52 oligomeric ring structure. Overall, <i>in vivo</i> and <i>in vitro</i> analyzes of these mutants indicate that Rad52 ring structure is important for protecting Rad51 filaments from Srs2, but can increase Rad51 filament stability and toxicity in Srs2-deficient cells. This stabilization function is distinct from Rad52 mediator and annealing activities.https://www.mdpi.com/2073-4409/10/6/1467genome stabilityDNA repairHomologous RecombinationRad52Rad51Srs2 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Emilie Ma Laurent Maloisel Léa Le Falher Raphaël Guérois Eric Coïc |
spellingShingle |
Emilie Ma Laurent Maloisel Léa Le Falher Raphaël Guérois Eric Coïc Rad52 Oligomeric N-Terminal Domain Stabilizes Rad51 Nucleoprotein Filaments and Contributes to Their Protection against Srs2 Cells genome stability DNA repair Homologous Recombination Rad52 Rad51 Srs2 |
author_facet |
Emilie Ma Laurent Maloisel Léa Le Falher Raphaël Guérois Eric Coïc |
author_sort |
Emilie Ma |
title |
Rad52 Oligomeric N-Terminal Domain Stabilizes Rad51 Nucleoprotein Filaments and Contributes to Their Protection against Srs2 |
title_short |
Rad52 Oligomeric N-Terminal Domain Stabilizes Rad51 Nucleoprotein Filaments and Contributes to Their Protection against Srs2 |
title_full |
Rad52 Oligomeric N-Terminal Domain Stabilizes Rad51 Nucleoprotein Filaments and Contributes to Their Protection against Srs2 |
title_fullStr |
Rad52 Oligomeric N-Terminal Domain Stabilizes Rad51 Nucleoprotein Filaments and Contributes to Their Protection against Srs2 |
title_full_unstemmed |
Rad52 Oligomeric N-Terminal Domain Stabilizes Rad51 Nucleoprotein Filaments and Contributes to Their Protection against Srs2 |
title_sort |
rad52 oligomeric n-terminal domain stabilizes rad51 nucleoprotein filaments and contributes to their protection against srs2 |
publisher |
MDPI AG |
series |
Cells |
issn |
2073-4409 |
publishDate |
2021-06-01 |
description |
Homologous recombination (HR) depends on the formation of a nucleoprotein filament of the recombinase Rad51 to scan the genome and invade the homologous sequence used as a template for DNA repair synthesis. Therefore, HR is highly accurate and crucial for genome stability. Rad51 filament formation is controlled by positive and negative factors. In <i>Saccharomyces cerevisiae,</i> the mediator protein Rad52 catalyzes Rad51 filament formation and stabilizes them, mostly by counteracting the disruptive activity of the translocase Srs2. Srs2 activity is essential to avoid the formation of toxic Rad51 filaments, as revealed by Srs2-deficient cells. We previously reported that Rad52 SUMOylation or mutations disrupting the Rad52–Rad51 interaction suppress Rad51 filament toxicity because they disengage Rad52 from Rad51 filaments and reduce their stability. Here, we found that mutations in Rad52 N-terminal domain also suppress the DNA damage sensitivity of Srs2-deficient cells. Structural studies showed that these mutations affect the Rad52 oligomeric ring structure. Overall, <i>in vivo</i> and <i>in vitro</i> analyzes of these mutants indicate that Rad52 ring structure is important for protecting Rad51 filaments from Srs2, but can increase Rad51 filament stability and toxicity in Srs2-deficient cells. This stabilization function is distinct from Rad52 mediator and annealing activities. |
topic |
genome stability DNA repair Homologous Recombination Rad52 Rad51 Srs2 |
url |
https://www.mdpi.com/2073-4409/10/6/1467 |
work_keys_str_mv |
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