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PreprintCited by 2DNA Repair MechanismsFungal and yeast genetics researchGenomics and Chromatin Dynamics

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Recruitment of Mre11 to recombination sites during meiosis

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Bouuaert, Corentin Claeys, Priyadarshini, Priyanka, Survi, Mahesh, Mouloud, Wael El Yazidi, Bohn, Regina, Ballet, Steven, Hunter, Neil, Volkov, Alexander·2025·Nature Communications
DOI 10.21203/rs.3.rs-7215871/v1PMC PMC12265730

Preprint — published as 10.1038/s41467-026-71310-5

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Abstract

The Mre11 nuclease is part of the highly conserved MRX complex involved in the repair of DNA double-strand breaks (DSBs). During meiosis in budding yeast, MRX is also required for the programmed induction of DSBs by Spo11, thereby initiating homologous recombination to promote accurate chromosome segregation. Recruitment of Mre11 to meiotic DSB sites depends on Rec114-Mei4 and Mer2 (RMM), which are thought to organize the meiotic DSB machinery by a mechanism involving biomolecular condensation. Here, we explored the role of Mre11 during meiosis and its relationship to RMM condensation. We show that both Mre11 and MRX complexes form DNA-dependent, hexanediol sensitive condensates in vitro . In vivo , Mre11 assembles into DNA damage-dependent foci in vegetative cells and DSB-independent foci in meiotic cells. In vitro condensates and in vivo foci both depend on the C-terminal intrinsically-disordered region (IDR) of Mre11. Importantly, while the Mre11 IDR is dispensable for vegetative DNA repair it is essential during meiosis. The C-terminal region of Mre11 forms a short α-helix that binds a conserved region of Mer2, and mutating residues within this interface reduces Mre11 foci and DSB formation. Finally, we identified a SUMO-interacting motif within the Mre11 IDR that enhances recruitment of Mre11 during meiosis and facilitates DSB formation. Our results provide new insights into the biophysical properties of Mre11 and its role in initiating meiotic recombination.

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Figure 1: Mre11 and the MRX complex assemble DNA-dependent condensates in vitro. (A, C) Effect of the presence of plasmid DNA (pUC19) on condensation of (A) Alexa488-labelled Mre11 and (C) eGFP-tagged MRX. (B, D) Effect of DNase I on (B) Mre11 and (D) MRX condensates. Condensates were pre-assembled for 20 minutes with 5.7 nM pUC19 prior to challenge. Quantification represents total foci intensity (green) in a field of view and total number of foci per 1000 μm2 (magenta). Foci intensity is normalized to the mean intensity of samples with DNA for A, C and untreated samples for B, D. Error bars represent mean ± SD from 8–12 fields of view. (E, F) Effect of the presence of 5% PEG-3350 on (E) Mre11 and (F) MRX condensate formation. Quantification represents total foci intensity (green) in a field of view (normalized to the mean of +PEG images) and total number of foci per 1000 μm2 (magenta). Error bars represent mean ± SD from 10–12 fields of view. (G, I) Effect of challenge with 5% 1,6-hexanediol and (H, J) 0.5 M NaCl on (G, H) Mre11 and (H, J) MRX condensates. Condensates were assembled for 20 minutes prior to challenge and incubated for 15 minutes and 10 minutes, respectively, after challenge. In the control reaction, an equivalent volume (1/20) of water was added instead of hexanediol. Quantification shows foci intensity (normalized to the mean of untreated samples) and total number of foci per 1000 μm2 (magenta). Error bars represent mean ± SD from 10–12 fields of view.
Figure 1: Mre11 and the MRX complex assemble DNA-dependent condensates in vitro. (A, C) Effect of the presence of plasmid DNA (pUC19) on condensation of (A) Alexa488-labelled Mre11 and (C) eGFP-tagged MRX. (B, D) Effect of DNase I on (B) Mre11 and (D) MRX condensates. Condensates were pre-assembled for 20 minutes with 5.7 nM pUC19 prior to challenge. Quantification represents total foci intensity (green) in a field of view and total number of foci per 1000 μm2 (magenta). Foci intensity is normalized to the mean intensity of samples with DNA for A, C and untreated samples for B, D. Error bars represent mean ± SD from 8–12 fields of view. (E, F) Effect of the presence of 5% PEG-3350 on (E) Mre11 and (F) MRX condensate formation. Quantification represents total foci intensity (green) in a field of view (normalized to the mean of +PEG images) and total number of foci per 1000 μm2 (magenta). Error bars represent mean ± SD from 10–12 fields of view. (G, I) Effect of challenge with 5% 1,6-hexanediol and (H, J) 0.5 M NaCl on (G, H) Mre11 and (H, J) MRX condensates. Condensates were assembled for 20 minutes prior to challenge and incubated for 15 minutes and 10 minutes, respectively, after challenge. In the control reaction, an equivalent volume (1/20) of water was added instead of hexanediol. Quantification shows foci intensity (normalized to the mean of untreated samples) and total number of foci per 1000 μm2 (magenta). Error bars represent mean ± SD from 10–12 fields of view.CC BY · PubMed Central
Figure 2: The C-terminal IDR of Mre11 is required for condensation. (A) AlphaFold2 predicted model of Mre11 (AF-P32829-F1-v4). (B) Domain structure of Mre11 and protein disorder prediction using IUPred server73. The ANCHOR score predicts the transition probability from a disordered to an ordered structure dependent on a binding partner. (C) Effect of deleting most of the phosphodiesterase domain (Δ10–270), or the capping domain and Rad50-binding domain (Δ290–472), (D) deleting the disordered region (ΔIDR, residues 524–677) and C-terminus (ΔC49, residues 644–692) on Mre11 condensation, and (E) deleting the C-terminus (ΔC49) on MRX condensation. Reactions in (C) and (D) contained 200 nM Mre11. Quantification represents total fluorescence intensity (green) in a field of view and total number of foci per 1000 μm2 (magenta). Foci intensity is normalized to the mean foci intensity of wild type Mre11 or MRX. Error bars represent mean ± SD from 10–14 fields of view for (C) and (D), and 21 fields of view for (E).
Figure 2: The C-terminal IDR of Mre11 is required for condensation. (A) AlphaFold2 predicted model of Mre11 (AF-P32829-F1-v4). (B) Domain structure of Mre11 and protein disorder prediction using IUPred server73. The ANCHOR score predicts the transition probability from a disordered to an ordered structure dependent on a binding partner. (C) Effect of deleting most of the phosphodiesterase domain (Δ10–270), or the capping domain and Rad50-binding domain (Δ290–472), (D) deleting the disordered region (ΔIDR, residues 524–677) and C-terminus (ΔC49, residues 644–692) on Mre11 condensation, and (E) deleting the C-terminus (ΔC49) on MRX condensation. Reactions in (C) and (D) contained 200 nM Mre11. Quantification represents total fluorescence intensity (green) in a field of view and total number of foci per 1000 μm2 (magenta). Foci intensity is normalized to the mean foci intensity of wild type Mre11 or MRX. Error bars represent mean ± SD from 10–14 fields of view for (C) and (D), and 21 fields of view for (E).CC BY · PubMed Central
Figure 3: The Mre11 IDR is required for vegetative foci formation but dispensable for DNA repair. (A) Effect of 1,6-hexanediol treatment on MMS-induced Mre11myc foci visualized by immunofluorescence analysis of vegetatively growing yeast nuclear spreads (B) Quantification of number of Mre11 foci (left) and Mre11 aggregates (right) before and after 1,6-hexanediol treatment. Error bar shows mean ± SD for at least 25 cells per strain. (C) Effect of Mre11myc truncations on MMS-induced foci formation as visualized by immunofluorescence. Error bars show mean ± SD of Mre11myc foci for WT (n = 51), mre11-ΔIDR (n = 52), and mre11-ΔC49 (n = 45). (D) Sensitivity of wild-type and truncated Mre11myc strains to methylmethanesulfonate (MMS) and camptothecin (CPT). Ten-fold serial dilutions from saturated cultures are shown, with dilutions on YPD plates as control.
Figure 3: The Mre11 IDR is required for vegetative foci formation but dispensable for DNA repair. (A) Effect of 1,6-hexanediol treatment on MMS-induced Mre11myc foci visualized by immunofluorescence analysis of vegetatively growing yeast nuclear spreads (B) Quantification of number of Mre11 foci (left) and Mre11 aggregates (right) before and after 1,6-hexanediol treatment. Error bar shows mean ± SD for at least 25 cells per strain. (C) Effect of Mre11myc truncations on MMS-induced foci formation as visualized by immunofluorescence. Error bars show mean ± SD of Mre11myc foci for WT (n = 51), mre11-ΔIDR (n = 52), and mre11-ΔC49 (n = 45). (D) Sensitivity of wild-type and truncated Mre11myc strains to methylmethanesulfonate (MMS) and camptothecin (CPT). Ten-fold serial dilutions from saturated cultures are shown, with dilutions on YPD plates as control.CC BY · PubMed Central
Figure 4: Mer2 recruits Mre11 during meiosis. (A) Effect of 1,6-hexanediol treatment on meiotic Mre11myc foci visualized by immunofluorescence analysis of yeast nuclear spreads 4 hours after transfer to SPM. Quantification shows mean and SD for untreated (n = 53) and 1,6-hexanediol treated cells (n = 73). (B, D) Immunofluorescence on meiotic nuclear spreads for myc-tagged (B) MRE11-WT, mre11-ΔC49, mre11-ΔIDR and (D) MRE11-WT in MER2, mer2Δ, and mer2-KRRR strains. Quantification shows mean and SD of Mre11myc foci for WT (n = 55), mre11-ΔC49 (n = 60), mre11-ΔIDR (n = 53), MER2 (n = 55), mer2Δ (n = 33), and mer2-KRRR (n = 57). (C) Spore viabilities of strains expressing wild-type or truncated Mre11myc. At least 22 tetrads were dissected for each strain (n ≥ 88 spores). (E) Quantification of sub-population of cells showing discrete Mre11 foci and aggregated Mre11 foci in indicated strains. Images used for analysis were from the same experiments in (B) and (D). (F) Colocalization of fluorescently-labelled Mer2 with wild-type or truncated Mre11. Reactions containing 200 nM of Alexa594Mer2 and eGFPMre11 were assembled separately for 10 minutes then mixed at 1:1 ratio for 30 minutes prior to imaging. Quantification shows the fraction of Mre11 foci overlapping Mer2 foci (white) in a field of view, normalized to Mre11-WT. Error bars represent mean ± SD from 10–15 fields of view. (G) Colocalization of fluorescently-labelled Mre11-C49 and Mer2. Condensates were assembled by mixing 800 nM of mScarletMre11-C49 and 200 nM of eGFPMer2 for 30 minutes prior to imaging. Controls reactions contained the same concentration but lacked either eGFPMer2 or mScarletMre11-C49, respectively.
Figure 4: Mer2 recruits Mre11 during meiosis. (A) Effect of 1,6-hexanediol treatment on meiotic Mre11myc foci visualized by immunofluorescence analysis of yeast nuclear spreads 4 hours after transfer to SPM. Quantification shows mean and SD for untreated (n = 53) and 1,6-hexanediol treated cells (n = 73). (B, D) Immunofluorescence on meiotic nuclear spreads for myc-tagged (B) MRE11-WT, mre11-ΔC49, mre11-ΔIDR and (D) MRE11-WT in MER2, mer2Δ, and mer2-KRRR strains. Quantification shows mean and SD of Mre11myc foci for WT (n = 55), mre11-ΔC49 (n = 60), mre11-ΔIDR (n = 53), MER2 (n = 55), mer2Δ (n = 33), and mer2-KRRR (n = 57). (C) Spore viabilities of strains expressing wild-type or truncated Mre11myc. At least 22 tetrads were dissected for each strain (n ≥ 88 spores). (E) Quantification of sub-population of cells showing discrete Mre11 foci and aggregated Mre11 foci in indicated strains. Images used for analysis were from the same experiments in (B) and (D). (F) Colocalization of fluorescently-labelled Mer2 with wild-type or truncated Mre11. Reactions containing 200 nM of Alexa594Mer2 and eGFPMre11 were assembled separately for 10 minutes then mixed at 1:1 ratio for 30 minutes prior to imaging. Quantification shows the fraction of Mre11 foci overlapping Mer2 foci (white) in a field of view, normalized to Mre11-WT. Error bars represent mean ± SD from 10–15 fields of view. (G) Colocalization of fluorescently-labelled Mre11-C49 and Mer2. Condensates were assembled by mixing 800 nM of mScarletMre11-C49 and 200 nM of eGFPMer2 for 30 minutes prior to imaging. Controls reactions contained the same concentration but lacked either eGFPMer2 or mScarletMre11-C49, respectively.CC BY · PubMed Central
Figure 5: The C-terminus of Mre11 binds a conserved motif of Mer2. (A) AlphaFold2 predicted model of Mre11 C-terminal residues (644–692, Mre11-C49) (orange) and Mer2 (41–100) (blue), aligned to the Mer2 coiled-coil domain (residues 41–247) (grey). Magnified-view shows amino acid residues at the predicted interaction surface, Mre11 residues L683, L686, and K690 (LLK) and Mer2 residues E50, Q54, E57, and K61 (EQEK). (B) Co-expression based pulldown assay between wild type and mutant MBPMre11-C49 (prey) and HisSUMOMer2 (bait). The LLK and EQEK mutants have the respective residues mutated to alanine. Asterisk (*) denotes free MBP. (C) Quantification of anti-MBP elution/input signal from pulldown in panel B, normalized to the wild type. Error bars represent mean ± SD from at least seven independent samples. (D) Immunofluorescence on meiotic nuclear spreads of Mre11myc in WT, mer2-EQEK, and mre11-ΔC15 strains. Quantification of Mre11myc foci in WT (n = 54), mer2-EQEK (n = 36), and mre11-ΔC15 (n = 38). (E, F) Southern blot analysis of meiotic DSB formation at the GAT1 hotspot. Quantification of panel F show mean and range from n = 2 experiments. (G) Spore viabilities of wild-type and mutant strains (n ≥ 88 spores).
Figure 5: The C-terminus of Mre11 binds a conserved motif of Mer2. (A) AlphaFold2 predicted model of Mre11 C-terminal residues (644–692, Mre11-C49) (orange) and Mer2 (41–100) (blue), aligned to the Mer2 coiled-coil domain (residues 41–247) (grey). Magnified-view shows amino acid residues at the predicted interaction surface, Mre11 residues L683, L686, and K690 (LLK) and Mer2 residues E50, Q54, E57, and K61 (EQEK). (B) Co-expression based pulldown assay between wild type and mutant MBPMre11-C49 (prey) and HisSUMOMer2 (bait). The LLK and EQEK mutants have the respective residues mutated to alanine. Asterisk (*) denotes free MBP. (C) Quantification of anti-MBP elution/input signal from pulldown in panel B, normalized to the wild type. Error bars represent mean ± SD from at least seven independent samples. (D) Immunofluorescence on meiotic nuclear spreads of Mre11myc in WT, mer2-EQEK, and mre11-ΔC15 strains. Quantification of Mre11myc foci in WT (n = 54), mer2-EQEK (n = 36), and mre11-ΔC15 (n = 38). (E, F) Southern blot analysis of meiotic DSB formation at the GAT1 hotspot. Quantification of panel F show mean and range from n = 2 experiments. (G) Spore viabilities of wild-type and mutant strains (n ≥ 88 spores).CC BY · PubMed Central
Figure 6: The Mre11 C-terminus contains a novel SUMO-interaction motif. (A) Schematic representation of SUMO-interacting motif (SIMs) in Mre11. (B, C) Average backbone amide chemical shift perturbations (Δδavg) of Smt3 in the presence of 1.2 molar equivalents of (B) wild type or (C) mutant SIM peptide (sequences indicated). The secondary structure of Smt3 is shown above the plots. The horizontal lines in (B) correspond to the average Δδavg (avg) plus one or two standard deviations (stdev). (D) Overlaid [1H,15N] HSQC spectra of Smt3 in its free form (black) or with 1.2 molar equivalents of wild type (green) or mutant (pink) SIM3 peptides. Blue arrows indicate residues that experience highest chemical shift. (E) Chemical shift mapping of the wild-type SIM peptide binding. Smt3 is shown as (top) cartoon and (bottom) molecular surface coloured by the Δδavg values (pink: Δδavg > avg + stdev; violet: Δδavg > avg + 2*stdev). The bound SIM peptide is in green. The disordered regions at Smt3 N- and C-termini are omitted for clarity. (F) Normalized heat per peak ΔH (kcal/mol) as a function of molar ratio (peptide/protein concentration) measured by isothermal titration calorimetry upon titration of wild-type (green) or mutant (pink) SIM peptide on Smt3. The solid line shows the best fit to a single binding site model for the wild-type peptide.
Figure 6: The Mre11 C-terminus contains a novel SUMO-interaction motif. (A) Schematic representation of SUMO-interacting motif (SIMs) in Mre11. (B, C) Average backbone amide chemical shift perturbations (Δδavg) of Smt3 in the presence of 1.2 molar equivalents of (B) wild type or (C) mutant SIM peptide (sequences indicated). The secondary structure of Smt3 is shown above the plots. The horizontal lines in (B) correspond to the average Δδavg (avg) plus one or two standard deviations (stdev). (D) Overlaid [1H,15N] HSQC spectra of Smt3 in its free form (black) or with 1.2 molar equivalents of wild type (green) or mutant (pink) SIM3 peptides. Blue arrows indicate residues that experience highest chemical shift. (E) Chemical shift mapping of the wild-type SIM peptide binding. Smt3 is shown as (top) cartoon and (bottom) molecular surface coloured by the Δδavg values (pink: Δδavg > avg + stdev; violet: Δδavg > avg + 2*stdev). The bound SIM peptide is in green. The disordered regions at Smt3 N- and C-termini are omitted for clarity. (F) Normalized heat per peak ΔH (kcal/mol) as a function of molar ratio (peptide/protein concentration) measured by isothermal titration calorimetry upon titration of wild-type (green) or mutant (pink) SIM peptide on Smt3. The solid line shows the best fit to a single binding site model for the wild-type peptide.CC BY · PubMed Central
Figure 7: A SUMO-SIM interaction fosters the recruitment of Mre11 during meiosis. (A) Immunofluorescence on meiotic nuclear spreads of myc-tagged MRE11-WT, mre11-SIM3, and mre11-SIM3+ΔC15 cells. Quantification of Mre11myc foci show mean ± SD of n = 50 cells per strain. (B) Southern blot analysis of meiotic DSB formation at GAT1 hotspot. Quantifications show mean ± range of n = 2 independent experiments. (C) Spore viabilities of wild-type and mutant MRE11 strains (n ≥ 88 spores). (D) Schematic model of the mechanisms that drive Mre11 recruitment during meiosis. Mre11 undergoes DNA-dependent condensation, driven by its IDR, and can mingle with Mer2 to form joint condensates. Mre11 is recruited to the DSB machinery via its C-terminus through a SUMO-SIM interaction and direct binding with Spo11 and Mer2.
Figure 7: A SUMO-SIM interaction fosters the recruitment of Mre11 during meiosis. (A) Immunofluorescence on meiotic nuclear spreads of myc-tagged MRE11-WT, mre11-SIM3, and mre11-SIM3+ΔC15 cells. Quantification of Mre11myc foci show mean ± SD of n = 50 cells per strain. (B) Southern blot analysis of meiotic DSB formation at GAT1 hotspot. Quantifications show mean ± range of n = 2 independent experiments. (C) Spore viabilities of wild-type and mutant MRE11 strains (n ≥ 88 spores). (D) Schematic model of the mechanisms that drive Mre11 recruitment during meiosis. Mre11 undergoes DNA-dependent condensation, driven by its IDR, and can mingle with Mer2 to form joint condensates. Mre11 is recruited to the DSB machinery via its C-terminus through a SUMO-SIM interaction and direct binding with Spo11 and Mer2.CC BY · PubMed Central

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Cites

Research works printed but not linked: 72

  1. Cooke MS, Evans MD, Dizdaroglu M, Lunec J, Oxidative (2003) DNA damage: mechanisms, mutation, and disease. FASEB j 17(10):1195–1214. 10.1096/fj.02-0752rev · DOI · Paper page
  2. De Almeida LC, Calil FA, Machado-Neto JA, Costa-Lotufo LV (2021) DNA damaging agents and DNA repair: From carcinogenesis to cancer therapy. Cancer Genet 252–253:6–24. 10.1016/j.cancergen.2020.12.002 · DOI
  3. Tubbs A, Nussenzweig A (2017) Endogenous DNA Damage as a Source of Genomic Instability in Cancer. Cell 168(4):644–656. 10.1016/j.cell.2017.01.002 · DOI · Paper page
  4. Oster S, Aqeilan RI, Programmed DNA (2020) Damage and Physiological DSBs: Mapping, Biological Significance and Perturbations in Disease States. Cells 9(8):1870. 10.3390/cells9081870 · DOI
  5. Börner GV, Hochwagen A, MacQueen AJ (2023) Meiosis in budding yeast. Genetics 225(2):iyad125. 10.1093/genetics/iyad125 · DOI
  6. Zickler D, Kleckner N, Meiosis (2023) Dances Between Homologs. Annu Rev Genet 57(1):1–63. 10.1146/annurev-genet-061323-044915 · DOI
  7. Lam I, Keeney S (2015) Mechanism and Regulation of Meiotic Recombination Initiation. Cold Spring Harb Perspect Biol 7(1):a016634. 10.1101/cshperspect.a016634 · DOI
  8. Yadav VK, Claeys Bouuaert C (2021) Mechanism and Control of Meiotic DNA Double-Strand Break Formation in S. cerevisiae. Front Cell Dev Biol 9:642737. 10.3389/fcell.2021.642737 · DOI
  9. Claeys Bouuaert C, Pu S, Wang J et al (2021) DNA-driven condensation assembles the meiotic DNA break machinery. Nature 592(7852):144–149. 10.1038/s41586-021-03374-w · DOI
  10. Claeys Bouuaert C, Tischfield SE, Pu S et al (2021) Structural and functional characterization of the Spo11 core complex. Nat Struct Mol Biol 28(1):92–102. 10.1038/s41594-020-00534-w · DOI · Paper page
  11. Johnson D, Crawford M, Cooper T et al (2021) Concerted cutting by Spo11 illuminates meiotic DNA break mechanics. Nature 594(7864):572–576. 10.1038/s41586-021-03389-3 · DOI
  12. Oger C, Claeys Bouuaert C (2025) SPO11 dimers are sufficient to catalyse DNA double-strand breaks in vitro. Nature 639(8055):792–799. 10.1038/s41586-024-08574-8 · DOI
  13. Kleckner N (2006) Chiasma formation: chromatin/axis interplay and the role(s) of the synaptonemal complex. Chromosoma 115(3):175–194. 10.1007/s00412-006-0055-7 · DOI
  14. Panizza S, Mendoza MA, Berlinger M et al (2011) Spo11-Accessory Proteins Link Double-Strand Break Sites to the Chromosome Axis in Early Meiotic Recombination. Cell 146(3):372–383. 10.1016/j.cell.2011.07.003 · DOI
  15. Alani E, Padmore R, Kleckner N (1990) Analysis of Wild-Type and rad50 mutants of Yeast Suggests an Intimate Relationship between Meiotic Chromosome Synapsis and Recombination. Cell 61(3):419–436. 10.1016/0092-8674(90)90524-I · DOI
  16. Ivanov EL, Korolevt VG, Fabre F (1992) Xrs2, a DNA Repair Gene of Saccharomyces Cerevisiae, Is Needed for Meiotic Recombination. Genetics 132(3):651–664. 10.1093/genetics/132.3.651 · DOI
  17. Nairz K, Klein F (1997) mre11S —a yeast mutation that blocks double-strand-break processing and permits nonhomologous synapsis in meiosis. Genes Dev. ;11(17):2272–2290. 10.1101/gad.11.17.2272 · DOI
  18. Keeney S (2001) Mechanism and control of meiotic recombination initiation. Curr Top Dev Biol 52:1–53. 10.1016/S0070-2153(01)52008-6 · DOI
  19. Moore JK, Haber JE (1996) Cell Cycle and Genetic Requirements of Two Pathways of Nonhomologous End-Joining Repair of Double-Strand Breaks in Saccharomyces cerevisiae. Mol Cell Biol 16(5):2164–2173. 10.1128/MCB.16.5.2164 · DOI · Paper page
  20. Paull TT, Gellert M (1998) The 3′ to 5′ Exonuclease Activity of Mre11 Facilitates Repair of DNA Double-Strand Breaks. Mol Cell 1(7):969–979. 10.1016/S1097-2765(00)80097-0 · DOI
  21. Garcia V, Phelps SEL, Gray S, Neale MJ (2011) Bidirectional resection of DNA double-strand breaks by Mre11 and Exo1. Nature 479(7372):241–244. 10.1038/nature10515 · DOI
  22. Casari E, Rinaldi C, Marsella A et al (2019) Processing of DNA Double-Strand Breaks by the MRX Complex in a Chromatin Context. Front Mol Biosci 6:43. 10.3389/fmolb.2019.00043 · DOI
  23. Gobbini E, Cassani C, Villa M, Bonetti D, Longhese M (2016) Functions and regulation of the MRX complex at DNA double-strand breaks. Microb Cell 3(8):329–337. 10.15698/mic2016.08.517 · DOI
  24. Paull TT (2018) 20 Years of Mre11 Biology: No End in Sight. Mol Cell 71(3):419–427. 10.1016/j.molcel.2018.06.033 · DOI
  25. Tisi R, Vertemara J, Zampella G, Longhese MP (2020) Functional and structural insights into the MRX/MRN complex, a key player in recognition and repair of DNA double-strand breaks. Comput Struct Biotechnol J 18:1137–1152. 10.1016/j.csbj.2020.05.013 · DOI
  26. Lamarche BJ, Orazio NI, Weitzman MD (2010) The MRN complex in double-strand break repair and telomere maintenance. FEBS Lett 584(17):3682–3695. 10.1016/j.febslet.2010.07.029 · DOI · Paper page
  27. Lu R, Zhang H, Jiang YN, Wang ZQ, Sun L, Zhou ZW (2021) Post-Translational Modification of MRE11: Its Implication in DDR and Diseases. Genes 12(8):1158. 10.3390/genes12081158 · DOI
  28. Oh J, Symington LS (2018) Role of the Mre11 Complex in Preserving Genome Integrity. Genes 9(12):589. 10.3390/genes9120589 · DOI
  29. Furuse M, Nagase Y, Tsubouchi H, Murakami-Murofushi K, Shibata T, Ohta K (1998) Distinct roles of two separable in vitro activities of yeast Mre11 in mitotic and meiotic recombination. EMBO J 17(21):6412–6425. 10.1093/emboj/17.21.6412 · DOI
  30. Usui T, Ohta T, Oshiumi H, Tomizawa J ichi, Ogawa H, Ogawa T (1998) Complex Formation and Functional Versatility of Mre11 of Budding Yeast in Recombination. Cell. ;95(5):705–716. 10.1016/S0092-8674(00)81640-2 · DOI
  31. Aithal R, Nangalia K, Spirek M, Chen D, Klein F, Krejci L (2024) Physical interaction with Spo11 mediates the localisation of Mre11 to chromatin in meiosis and promotes its nuclease activity. Nucleic Acids Res 52(8):4328–4343. 10.1093/nar/gkae111 · DOI
  32. Wen Q, Scorah J, Phear G, Rodgers G, Rodgers S, Meuth M (2008) A Mutant Allele of MRE11 Found in Mismatch Repair-deficient Tumor Cells Suppresses the Cellular Response to DNA Replication Fork Stress in a Dominant Negative Manner. Cohen-Fix O, ed. MBoC. ;19(4):1693–1705. 10.1091/mbc.e07-09-0975 · DOI
  33. Tamai T, Reginato G, Ojiri R et al (2024) Sae2 controls Mre11 endo- and exonuclease activities by different mechanisms. Nat Commun 15(1):7221. 10.1038/s41467-024-51493-5 · DOI
  34. Kissling VM, Reginato G, Bianco E et al (2022) Mre11-Rad50 oligomerization promotes DNA double-strand break repair. Nat Commun 13(1):2374. 10.1038/s41467-022-29841-0 · DOI
  35. Kroschwald S, Maharana S, Simon A (2017) Hexanediol: a chemical probe to investigate the material properties of membrane-less compartments. Matters Published online May 22. 10.19185/matters.201702000010 · DOI
  36. Lammens K, Bemeleit DJ, Möckel C et al (2011) The Mre11:Rad50 Structure Shows an ATP-Dependent Molecular Clamp in DNA Double-Strand Break Repair. Cell 145(1):54–66. 10.1016/j.cell.2011.02.038 · DOI
  37. Rotheneder M, Stakyte K, Van De Logt E et al (2023) Cryo-EM structure of the Mre11-Rad50-Nbs1 complex reveals the molecular mechanism of scaffolding functions. Mol Cell 83(2):167–185e9. 10.1016/j.molcel.2022.12.003 · DOI
  38. Petrini J, Hohl M, Yu Y, Kuryavyi V, Patel D (2024) Structure guided functional analysis of the S. cerevisiae Mre11 complex. Published online Dec 9. 10.21203/rs.3.rs-5390974/v1 · DOI
  39. Bergerat A, De Massy B, Gadelle D, Varoutas PC, Nicolas A, Forterre P (1997) An atypical topoisomerase II from archaea with implications for meiotic recombination. Nature 386(6623):414–417. 10.1038/386414a0 · DOI
  40. Borde V, Lin W, Novikov E, Petrini JH, Lichten M, Nicolas A (2004) Association of Mre11p with Double-Strand Break Sites during Yeast Meiosis. Mol Cell 13(3):389–401. 10.1016/S1097-2765(04)00034-6 · DOI
  41. Rousová D, Nivsarkar V, Altmannova V et al (2021) Novel mechanistic insights into the role of Mer2 as the keystone of meiotic DNA break formation. eLife 10:e72330. 10.7554/eLife.72330 · DOI
  42. Daccache D, De Jonge E, Liloku P et al (2023) Evolutionary conservation of the structure and function of meiotic Rec114 – Mei4 and Mer2 complexes. Genes Dev 37(11–12):535–553. 10.1101/gad.350462.123 · DOI
  43. Tessé S, Bourbon HM, Debuchy R et al (2017) Asy2/Mer2: an evolutionarily conserved mediator of meiotic recombination, pairing, and global chromosome compaction. Genes Dev 31(18):1880–1893. 10.1101/gad.304543.117 · DOI
  44. Bhagwat NR, Owens SN, Ito M et al (2021) SUMO is a pervasive regulator of meiosis. eLife 10:e57720. 10.7554/eLife.57720 · DOI
  45. Chen YJ, Chuang YC, Chuang CN et al (2016) S. cerevisiae Mre11 recruits conjugated SUMO moieties to facilitate the assembly and function of the Mre11-Rad50-Xrs2 complex. Nucleic Acids Res 44(5):2199–2213. 10.1093/nar/gkv1523 · DOI
  46. Zhao Q, Xie Y, Zheng Y et al (2014) GPS-SUMO: a tool for the prediction of sumoylation sites and SUMO-interaction motifs. Nucleic Acids Res 42(W1):W325–W330. 10.1093/nar/gku383 · DOI
  47. Lascorz J, Codina-Fabra J, Reverter D, Torres-Rosell J (2022) SUMO-SIM interactions: From structure to biological functions. Semin Cell Dev Biol 132:193–202. 10.1016/j.semcdb.2021.11.007 · DOI
  48. Song J, Zhang Z, Hu W, Chen Y (2005) Small Ubiquitin-like Modifier (SUMO) Recognition of a SUMO Binding Motif. J Biol Chem 280(48):40122–40129. 10.1074/jbc.M507059200 · DOI
  49. Keusekotten K, Bade VN, Meyer-Teschendorf K et al (2014) Multivalent interactions of the SUMO-interaction motifs in RING finger protein 4 determine the specificity for chains of the SUMO. Biochem J 457(1):207–214. 10.1042/BJ20130753 · DOI
  50. Symington LS (2016) Mechanism and regulation of DNA end resection in eukaryotes. Crit Rev Biochem Mol Biol 51(3):195–212. 10.3109/10409238.2016.1172552 · DOI
  51. Gnügge R, Symington LS (2017) Keeping it real: MRX–Sae2 clipping of natural substrates. Genes Dev 31(23–24):2311–2312. 10.1101/gad.310771.117 · DOI
  52. Chin GM, Villeneuve AM (2001) C. elegans mre-11 is required for meiotic recombination and DNA repair but is dispensable for the meiotic G2 DNA damage checkpoint. Genes Dev 15(5):522–534. 10.1101/gad.864101 · DOI
  53. Puizina J, Siroky J, Mokros P, Schweizer D, Riha K (2004) Mre11 deficiency in Arabidopsis is associated with chromosomal instability in somatic cells and Spo11-dependent genome fragmentation during meiosis. Plant Cell 16(8):1968–1978. 10.1105/tpc.104.022749 · DOI
  54. Young JA, Hyppa RW, Smith GR (2004) Conserved and nonconserved proteins for meiotic DNA breakage and repair in yeasts. Genetics 167(2):593–605. 10.1534/genetics.103.023762 · DOI
  55. Liu Y, Lin Z, Yan J, Zhang X, Tong MH (2024) A Rad50-null mutation in mouse germ cells causes reduced DSB formation, abnormal DSB end resection and complete loss of germ cells. Development 151(8). 10.1242/dev.202312 · DOI
  56. Kim S, Yamada S, Li T et al (2025) Mouse MRE11-RAD50-NBS1 is needed to start and extend meiotic DNA end resection. Nat Commun 16(1). 10.1038/s41467-025-57928-x · DOI
  57. Hayashi M, Chin GM, Villeneuve AM (2007) C. elegans germ cells switch between distinct modes of double-strand break repair during meiotic prophase progression. PLoS Genet 3(11). https://doi.org/10.1371/journal.pgen.0030191 · DOI
  58. Girard C, Roelens B, Zawadzki KA, Villeneuve AM (2018) Interdependent and separable functions of Caenorhabditis elegans MRN-C complex members couple formation and repair of meiotic DSBs. Proc Natl Acad Sci USA 115(19). 10.1073/pnas.1719029115 · DOI
  59. Bishop K, Kleckner N, DMCI (1992) A meiosis-specific yeast homolog of E. coli recA required for recombination, synaptonemal complex formation, and cell cycle progression. Cell 69(3):439–456. 10.1016/0092-8674(92)90446-j · DOI · Paper page
  60. Tran PT, Erdeniz N, Dudley S, Liskay RM (2002) Characterization of nuclease-dependent functions of Exo1p in Saccharomyces cerevisiae. DNA Repair 1(11):895–912. 10.1016/s1568-7864(02)00114-3 · DOI · Paper page
  61. Joshi N, Brown MS, Bishop DK, Börner GV (2015) Gradual implementation of the meiotic recombination program via checkpoint pathways controlled by global DSB levels. Mol Cell 57(5):797–811. 10.1016/j.molcel.2014.12.027 · DOI
  62. Mimitou EP, Yamada S, Keeney S (2017) A global view of meiotic double-strand break end resection. Science 335(6320):40–45. 10.1126/science.aak9704 · DOI
  63. Li J, Hooker GW, Roeder GS (2006) Saccharomyces cerevisiae Mer2, Mei4 and Rec114 form a complex required for meiotic double-strand break formation. Genetics 173(4):1969–1981. 10.1534/genetics.106.058768 · DOI
  64. Henderson KA, Kee K, Maleki S, Santini PA, Keeney S (2006) Cyclin-Dependent Kinase directly regulates initiation of meiotic recombination. Cell 125(7):1321–1332. 10.1016/j.cell.2006.04.039 · DOI
  65. Maleki S, Neale MJ, Arora C, Henderson KA, Keeney S (2007) Interactions between Mei4, Rec114, and other proteins required for meiotic DNA double-strand break formation in Saccharomyces cerevisiae. Chromosoma 116(5):471–486. 10.1007/s00412-007-0111-y · DOI
  66. Yunus AA, Lima CD (2009) Purification of SUMO Conjugating Enzymes and Kinetic Analysis of Substrate Conjugation. In: Ulrich HD (ed) SUMO Protocols. Methods in Molecular Biology, vol 497. Humana, pp 167–186. doi:10.1007/978-1-59745-566-4_11 · DOI
  67. Cannavo E, Cejka P, Kowalczykowski SC (2013) Relationship of DNA degradation by Saccharomyces cerevisiae Exonuclease 1 and its stimulation by RPA and Mre11-Rad50-Xrs2 to DNA end resection. Proc Natl Acad Sci USA 110(18). 10.1073/pnas.1305166110 · DOI
  68. Murakami H, Borde V, Nicolas A, Keeney S (2009) Gel electrophoresis assays for analyzing DNA double-strand breaks in Saccharomyces cerevisiae at various spatial resolutions. In: Keeney S (ed) Meiosis. Methods in Molecular Biology, vol 557. Humana, pp 117–142. doi:10.1007/978-1-59745-527-5_9 · DOI
  69. Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A, NMRPipe (1995) A multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6(3). 10.1007/BF00197809 · DOI
  70. Vranken WF, Boucher W, Stevens TJ et al (2005) The CCPN data model for NMR spectroscopy: Development of a software pipeline. Proteins 59(4):687–696. 10.1002/prot.20449 · DOI
  71. Sheng W, Liao X (2002) Solution structure of a yeast ubiquitin-like protein Smt3: The role of structurally less defined sequences in protein–protein recognitions. Protein Sci 11(6):1482–1491. 10.1110/ps.0201602 · DOI
  72. Dosztanyi Z, Csizmok V, Tompa P, Simon I (2005) IUPred: web server for the prediction of intrinsically unstructured regions of proteins based on estimated energy content. Bioinformatics 21(16):3433–3434. 10.1093/bioinformatics/bti541 · DOI

Metadata

Authors
Bouuaert, C. C., Priyadarshini, P., Survi, M., Mouloud, W. E. Y., Bohn, R., Ballet, S., Hunter, N., & Volkov, A.
Affiliations
  • UCLouvain (BE)
  • Vrije Universiteit Brussel (BE)
  • Howard Hughes Medical Institute / UC Davis
  • Howard Hughes Medical Institute (US)
Corresponding author
  • Bouuaert, Corentin Claeys
  • Priyadarshini, Priyanka
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2025-08-19
Volume
17
Issue
1
ISSN
2041-1723 (ISSN-L)
DOI
10.21203/rs.3.rs-7215871/v1
PMC
PMC12265730
OpenAlex
W4412172758
Cited by
2
References
1
Citations by year
2026: 2
FWCI
4.13
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • Research Square (acceptedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed Central (submittedVersion, cc-by)
  • DIAL (Catholic University of Leuven) (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Université Catholique de Louvain
  • European Commission (HORIZON2020)
  • Fonds De La Recherche Scientifique - FNRS
  • Vrije Universiteit Brussel
  • National Institutes of Health
  • National Institute of General Medical Sciences (R01GM074223)
  • H2020 European Research Council (802525)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Keywords
MRE11, MRX complex, meiotic recombination, DNA double-strand break repair, Spo11, biomolecular condensates, intrinsically disordered regions, SUMO-interacting motif, DNA damage foci, Saccharomyces cerevisiae

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