FIGL1 attenuates meiotic interhomolog repair and is counteracted by RAD51 paralog XRCC2 and chromosome axis protein ASY1 during meiosis DOI

Côme Emmenecker,

Simine Pakzad,

Fatou Ture

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: April 28, 2024

Abstract Two recombinases, RAD51 and DMC1, catalyze meiotic break repair to ensure crossovers (COs) between homologous chromosomes (interhomolog) rather than sisters (intersister). FIDGETIN-LIKE-1 (FIGL1) downregulates both recombinases. However, the understanding of FIGL1 functions in remains limited. Here, we discover new genetic interactions Arabidopsis thaliana that are important vivo determinants outcome. In figl1 , compromising RAD51-dependent by either losing paralogs (RAD51B or XRCC2) RAD54 inhibiting RAD51’s catalytic activity results unrepaired breaks CO defects. Further, XRCC2 physically interacts with partially counteracts for focus formation. Our data support RAD51-mediated mechanisms compensate dysfunction. is dispensable intersister dmc1 but essential completion mutants impaired DMC1 interhomolog bias such as asy1 . We show attenuates repair, ASY1 promote recombination.

Language: Английский

Promotion of DNA end resection by BRCA1–BARD1 in homologous recombination DOI
Sameer Salunkhe, James M. Daley, Hardeep Kaur

et al.

Nature, Journal Year: 2024, Volume and Issue: 634(8033), P. 482 - 491

Published: Sept. 11, 2024

Language: Английский

Citations

8
Positive and negative regulators of RAD51/DMC1 in homologous recombination and DNA replication DOI
Masaru Ito, Yurika Fujita, Akira Shinohara

et al.

DNA repair, Journal Year: 2023, Volume and Issue: 134, P. 103613 - 103613

Published: Dec. 13, 2023

Language: Английский

Citations

14
The human Shu complex promotes RAD51 activity by modulating RPA dynamics on ssDNA DOI Creative Commons
Sarah R Hengel, Katherine G. Oppenheimer, Chelsea M. Smith

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Aug. 21, 2024

Templated DNA repair that occurs during homologous recombination and replication stress relies on RAD51. RAD51 activity is positively regulated by BRCA2 the paralogs. The Shu complex a paralog-containing consisting of SWSAP1, SWS1, SPIDR. We demonstrate SWSAP1-SWS1 binds RAD51, maintains filament stability, enables strand exchange. Using single-molecule confocal fluorescence microscopy combined with optical tweezers, we show decorates filaments proficient for recombination. also find enhances RPA diffusion ssDNA. Importantly, human sgSWSAP1 sgSWS1 knockout cells are sensitive to pharmacological inhibition PARP APE1. Lastly, identify cancer variants in SWSAP1 alter formation. Together, stimulates RAD51-dependent high-fidelity may be an important new therapeutic target. promotes regulating Here authors reveal proteins, SWSAP1-SWS1, decorate ssDNA facilitate its exchange reaction stimulating knockouts Olaparib sensitive.

Language: Английский

Citations

4
RAD51 paralogs synergize with RAD51 to protect reversed forks from cellular nucleases DOI Creative Commons

Chia-Lun Guh,

Kai‐Hang Lei, Yi‐An Chen

et al.

Nucleic Acids Research, Journal Year: 2023, Volume and Issue: 51(21), P. 11717 - 11731

Published: Oct. 16, 2023

Abstract Fork reversal is a conserved mechanism to prevent stalled replication forks from collapsing. Formation and protection of reversed are two crucial steps in ensuring fork integrity stability. Five RAD51 paralogs, namely, RAD51B, RAD51C, RAD51D, XRCC2 XRCC3, which share sequence structural similarity the recombinase RAD51, play poorly defined mechanistic roles these processes. Here, using purified BCDX2 (RAD51BCD-XRCC2) CX3 (RAD51C-XRCC3) complexes vitro reconstituted biochemical systems, we mechanistically dissect their functions forming protecting forks. We show that both paralog lack activities. Whereas exhibits modest activity, significantly synergizes with protect DNA against attack by nucleases MRE11 EXO1. contingent upon ability form functional nucleoprotein filament on DNA. Collectively, our results provide evidence for hitherto unknown function paralogs synergizing degradation stressed

Language: Английский

Citations

8
Human RAD52 stimulates the RAD51-mediated homology search DOI Creative Commons

Ali Akbar Muhammad,

Clara Basto,

Thibaut Peterlini

et al.

Life Science Alliance, Journal Year: 2023, Volume and Issue: 7(3), P. e202201751 - e202201751

Published: Dec. 11, 2023

Homologous recombination (HR) is a DNA repair mechanism of double-strand breaks and blocked replication forks, involving process homology search leading to the formation synaptic intermediates that are regulated ensure genome integrity. RAD51 recombinase plays central role in this mechanism, supported by its RAD52 BRCA2 partners. If mediator function load on RPA-ssDNA well established, HR still far from understood. We used transmission electron microscopy combined with biochemistry characterize sequential participation RPA, RAD52, assembly filament activity. Although our results confirm lacks activity, can tightly bind RPA-coated ssDNA, inhibit activity BRCA2, form shorter RAD51-RAD52 mixed filaments more efficient complexes D-loops, resulting frequent multi-invasions as well. situ interaction between after break induction vivo. This study provides new molecular insights into regulation presynaptic during human HR.

Language: Английский

Citations

8
The human Shu complex promotes RAD51 activity by modulating RPA dynamics on ssDNA DOI Open Access
Sarah R Hengel,

Katherine Oppenheimer,

Chelsea M. Smith

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Feb. 15, 2024

Abstract Templated DNA repair that occurs during homologous recombination and replication stress relies on RAD51. RAD51 activity is positively regulated by BRCA2 the paralogs. The Shu complex a paralog-containing consisting of SWSAP1 SWS1. We demonstrate SWSAP1-SWS1 binds RAD51, maintains filament stability, enables strand exchange. Using single molecule confocal fluorescence microscopy combined with optical tweezers, we show decorates filaments proficient for recombination. also find enhances RPA diffusion ssDNA. Importantly, human sgSWSAP1 sgSWS1 knockout cells are sensitive to pharmacological inhibition PARP APE1. Lastly, identify cancer variants in alter SWS1 formation. Together, stimulates RAD51-dependent high-fidelity may be an important new therapeutic target.

Language: Английский

Citations

2
High-resolution functional mapping of RAD51C by saturation genome editing DOI Creative Commons

Rebeca Olvera-León,

Fang Zhang, Victoria Offord

et al.

Cell, Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 1, 2024

Pathogenic variants in RAD51C confer an elevated risk of breast and ovarian cancer, while individuals homozygous for specific alleles may develop Fanconi anemia. Using saturation genome editing (SGE), we functionally assess 9,188 unique variants, including >99.5% all possible coding sequence single-nucleotide alterations. By computing changes variant abundance Gaussian mixture modeling (GMM), classify 3,094 to be disruptive use clinical truth sets reveal accuracy/concordance classification >99.9%. Cell fitness was the primary assay readout allowing us observe a phenomenon where missense exhibit distinct depletion kinetics potentially suggesting that they represent hypomorphic alleles. We further explored our exhaustive functional map, revealing critical residues on structure resolving found cancer-segregating kindred. Furthermore, through interrogation UK Biobank large multi-center cancer cohort, find significant associations between SGE-depleted diagnoses.

Language: Английский

Citations

2
Molecular basis of FIGNL1 in dissociating RAD51 from DNA and chromatin DOI
Alexander Carver, Tai-Yuan Yu, Luke A. Yates

et al.

Science, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 5, 2024

Maintaining genome integrity is an essential and challenging process. RAD51 recombinase, the central player of several crucial processes in repairing DNA protecting integrity, forms filaments on DNA, which are tightly regulated. One these regulators FIGNL1, that prevents persistent foci without or after damage genotoxic chromatin association cells. The cryogenic electron microscopy structure FIGNL1 complex with reveals a non-planar hexamer N terminus enclosure pore. Mutations pore loop catalytic residues render it defective filament disassembly lethal mouse embryonic stem Our study unique mechanism for removing from bound substrates provides molecular basis maintaining stability.

Language: Английский

Citations

2
Photonic Crystal Surface Mode Real-Time Imaging of RAD51 DNA Repair Protein Interaction with the ssDNA Substrate DOI Creative Commons
Galina Nifontova,

Cathy Charlier,

Nizar Ayadi

et al.

Biosensors, Journal Year: 2024, Volume and Issue: 14(1), P. 43 - 43

Published: Jan. 14, 2024

Photonic crystals (PCs) are promising tools for label-free sensing in drug discovery screening, diagnostics, and analysis of ligand–receptor interactions. Imaging PC surface modes has emerged as a novel approach to the detection multiple binding events at sensor surface. modification decoration with recognition units yield an interface providing highly sensitive cancer biomarkers, antibodies, oligonucleotides. The RAD51 protein plays central role DNA repair via homologous recombination pathway. This recombinase is essential genome stability its overexpression often correlated aggressive cancer. therefore potential target therapeutic strategy Here, we report designing PC-based array real-time monitoring oligonucleotide–RAD51 recruitment by means mode imaging validation concept this approach. Our data demonstrate that designed biosensor ensures multiplexed association–dissociation biomarker damage using microfluidic array. obtained results highlight developed technique testing functionality candidate drugs, discovering new molecular targets entities. paves way further adaption bioanalytical use high-content screening identify inhibitor drugs targeting nucleoprotein filament or discover targets.

Language: Английский

Citations

1
All who wander are not lost: the search for homology during homologous recombination DOI Creative Commons
Jingyi Hu, J. Brooks Crickard

Biochemical Society Transactions, Journal Year: 2024, Volume and Issue: 52(1), P. 367 - 377

Published: Feb. 7, 2024

Homologous recombination (HR) is a template-based DNA double-strand break repair pathway that functions to maintain genomic integrity. A vital component of the HR reaction identification template be used during repair. This occurs through mechanism known as homology search. The search in two steps: collision step which pieces are forced collide and selection results homologous pairing between matching sequences. Selection facilitated by recombinases RecA/Rad51 family proteins cooperation with helicases, translocases, topoisomerases determine overall fidelity match. menagerie molecular machines acts regulate critical intermediates These include recombinase filaments probe for short stretches early strand invasion form displacement loops (D-loops) stabilize paired DNA. Here, we will discuss recent advances understanding how these specific regulated on level reaction. We also stability influences ultimate outcomes Finally, physiological models developed explain protects genome.

Language: Английский

Citations

1