Rad52 Acts as an Assembly Chaperone to Form and Stabilize Rad51 Filaments Through a Large C-Terminus 85-Residue Segment DOI Open Access
Émilie Ma,

Fadma Lakhal,

Eleni Litsardaki

et al.

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

Published: Dec. 5, 2024

Abstract Homologous recombination (HR) is essential for the repair of DNA double-strand breaks and restart stalled replication forks. A critical step in HR formation Rad51 nucleofilaments, which perform homology search strand invasion a homologous sequence required synthesis. In yeast Saccharomyces cerevisiae , Rad52 facilitates nucleofilament by mediating loading onto ssDNA counteracting dissociation filaments translocase Srs2. The molecular basis these two functions remains unclear. Our integrative structural analyses Rad51-Rad52 interaction, combining NMR, SAXS, modeling, reveal that an 85-residue segment Rad52, conserved fungi, folds upon binding to broad surface monomer. Notably, it includes FxxA motif BRC repeats BRCA2 at Rad51-Rad51 interface. This mode was validated through extensive set mutations. Using vivo assays functional fluorescent GFP-Rad51 fusion protein, we demonstrated this entire filament formation. These findings highlight how as assembly chaperone, preventing oligomerization, promoting nucleation nucleofilaments on ssDNA, protecting from destabilization

Language: Английский

Single-molecule fluorescence imaging of DNA maintenance protein binding dynamics and activities on extended DNA DOI
Elizabeth Marie Irvin, Hong Wang

Current Opinion in Structural Biology, Journal Year: 2024, Volume and Issue: 87, P. 102863 - 102863

Published: June 15, 2024

Language: Английский

Citations

2
Mei5–Sae3 stabilizes Dmc1 nucleating clusters for efficient Dmc1 assembly on RPA-coated single-stranded DNA DOI Creative Commons

Chin-Dian Wei,

Hao-Yen Chang,

Chia-Hua Lu

et al.

Nucleic Acids Research, Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 14, 2024

Abstract Interhomolog recombination in meiosis requires a meiosis-specific recombinase, Dmc1. In Saccharomyces cerevisiae, the Mei5–Sae3 complex facilitates loading of Dmc1 onto replication protein A (RPA)-coated single-stranded DNA (ssDNA) to form nucleoprotein filaments. vivo, and are interdependent their colocalization on chromosomes. However, mechanistic role mediating activity remains unclear. We used single-molecule fluorescence resonance energy transfer spectroscopy experiments elucidate how stimulates assembly ssDNA RPA-coated ssDNA. showed that stabilized nucleating clusters with two three molecules naked by preferentially reducing dissociation rates. also stimulated DNA. Using green fluorescent protein-labeled RPA, we coexistence an intermediate RPA before dissociation. Moreover, displacement efficiency depended concentration, its dependence was positively correlated stability short These findings suggest molecular model mediates binding stabilizing clusters, thus altering dynamics promote

Language: Английский

Citations

0
The role of RAD51 regulators and variants in primary ovarian insufficiency, endometriosis, and polycystic ovary syndrome DOI Creative Commons
Maggie Witham, Sarah R Hengel

Deleted Journal, Journal Year: 2024, Volume and Issue: 1(4)

Published: Sept. 28, 2024

Abstract The study of RAD51 regulators in female reproductive diseases has novel biomarker potential and implications for therapeutic advancement. Regulators play important roles maintaining genome integrity variations these genes have been identified including primary ovarian insufficiency (POI), endometriosis, polycystic ovary syndrome (PCOS). modulators change activity homologous recombination, replication stress, template switching pathways. However, molecular proteins insufficiency, understudied. For each disease, we provide its definition, current diagnostic treatment strategies, associated genetic variations. Variants were discovered RAD51, DMC1, RAD51B, SWS1, SPIDR, XRCC2 BRCA2 linked with POI. Endometriosis is variants XRCC3, BRCA1 CSB genes. PCOS. Our analysis biomarkers POI (DMC1 RAD51B) PCOS (BRCA1). Further biochemical cellular analyses regulator functions disorders will advance our understanding the pathogenesis diseases.

Language: Английский

Citations

0
Rad52 Acts as an Assembly Chaperone to Form and Stabilize Rad51 Filaments Through a Large C-Terminus 85-Residue Segment DOI Open Access
Émilie Ma,

Fadma Lakhal,

Eleni Litsardaki

et al.

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

Published: Dec. 5, 2024

Abstract Homologous recombination (HR) is essential for the repair of DNA double-strand breaks and restart stalled replication forks. A critical step in HR formation Rad51 nucleofilaments, which perform homology search strand invasion a homologous sequence required synthesis. In yeast Saccharomyces cerevisiae , Rad52 facilitates nucleofilament by mediating loading onto ssDNA counteracting dissociation filaments translocase Srs2. The molecular basis these two functions remains unclear. Our integrative structural analyses Rad51-Rad52 interaction, combining NMR, SAXS, modeling, reveal that an 85-residue segment Rad52, conserved fungi, folds upon binding to broad surface monomer. Notably, it includes FxxA motif BRC repeats BRCA2 at Rad51-Rad51 interface. This mode was validated through extensive set mutations. Using vivo assays functional fluorescent GFP-Rad51 fusion protein, we demonstrated this entire filament formation. These findings highlight how as assembly chaperone, preventing oligomerization, promoting nucleation nucleofilaments on ssDNA, protecting from destabilization

Language: Английский

Citations

0