Analysis of rad-51 separation of function allele suggests
divergence of the synthesis-dependent strand annealing and
double Holliday junction pathways prior to RAD-51 filament
disassembly

J 2025

Analysis of rad-51 separation of function allele suggests divergence of the synthesis-dependent strand annealing and double Holliday junction pathways prior to RAD-51 filament disassembly

OBERLITNER, Joseph; Maggie TINMAN; Aasthika DAS; Emily KOURY; Nicola SILVA et al.

Základní údaje

Originální název

Analysis of rad-51 separation of function allele suggests divergence of the synthesis-dependent strand annealing and double Holliday junction pathways prior to RAD-51 filament disassembly

Autoři

OBERLITNER, Joseph; Maggie TINMAN; Aasthika DAS; Emily KOURY; Nicola SILVA a Sarit SMOLIKOVE

Vydání

Genetics, CARY, OXFORD UNIV PRESS, 2025, 0016-6731

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10603 Genetics and heredity

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 5.100 v roce 2024

Označené pro přenos do RIV

Ano

Organizační jednotka

Lékařská fakulta

Klíčová slova anglicky

C. elegans; meiosis; RAD51; crossover

Štítky

14110513, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 6. 8. 2025 10:40, Mgr. Tereza Miškechová

Anotace

V originále

DNA double-strand breaks (DSBs) are formed in meiosis, so their repair in the homologous recombination (HR) pathway will lead to crossover formation, which is essential for successful chromosome segregation. HR contains 2 subpathways: synthesis-dependent strand annealing (SDSA) that creates noncrossover and double Holliday junction (dHJ) that generates crossovers. RAD-51 is a protein essential to the formation of all products of HR, as it assembles on the processed DSB, allowing the invasion of the single-stranded DNA into a region of homology. RAD-51 is removed by RAD-54.L after invasion to allow for repair to occur. Here, we investigate a separation of function allele of rad-51, rad-51::FLAG, as compared to 2 other RAD-51 alleles: rad-51::degron and GFP::rad-51. rad-51::FLAG displays slowed repair kinetics, resulting in an accumulation of RAD-51 foci. rad-51::FLAG worms also activate the DSB checkpoint, but to a less extant than that of rad-51 null mutants. In a proximity ligation assay, RAD-54.L and RAD-51 show enriched colocalization in rad-51::FLAG germlines (but not in rad-51::degron), consistent with stalling at the strand invasion step in HR. The defects in RAD-51 disassembly in rad-51::FLAG mutants lead to formation of chromosomal fragments, similar in their magnitude to ones observed in rad-51 or rad-54.L null mutants. However, rad-51::FLAG mutants (unlike a rad-51 null, GFP::rad-51 or rad-54.L null mutants) displayed no defects in the formation of crossover-designated sites (via GFP::COSA-1 localization). Given that rad-51::FLAG worms show checkpoint activation and chromosomal fragments, these results suggest that crossover repair concludes normally, while the noncrossover pathway is perturbed. This is strikingly different from rad-51::degron and GFP::rad-51 strains, which are proficient or deficient in both pathways, respectively. These results suggest that noncrossovers vs crossovers have distinct recombination intermediates and diverge prior to RAD-51 disassembly.

Návaznosti

GA23-04918S, projekt VaV

Název: Mechanismus založený na modifikaci chromatinu osvětluje novou dráhu pro vytvoření meiotické synapse chromozomů

Investor: Grantová agentura ČR, Mechanismus založený na modifikaci chromatinu osvětluje novou dráhu pro vytvoření meiotické synapse chromozomů

Citovat

OBERLITNER, Joseph; Maggie TINMAN; Aasthika DAS; Emily KOURY; Nicola SILVA a Sarit SMOLIKOVE. Analysis of rad-51 separation of function allele suggests divergence of the synthesis-dependent strand annealing and double Holliday junction pathways prior to RAD-51 filament disassembly. Genetics. CARY: OXFORD UNIV PRESS, 2025, roč. 230, č. 2, s. 1-16. ISSN 0016-6731. Dostupné z: https://doi.org/10.1093/genetics/iyaf063.

@article{2511225,
   author = {Oberlitner, Joseph and Tinman, Maggie and Das, Aasthika and Koury, Emily and Silva, Nicola and Smolikove, Sarit},
   article_location = {CARY},
   article_number = {2},
   doi = {https://doi.org/10.1093/genetics/iyaf063},
   keywords = {C. elegans; meiosis; RAD51; crossover},
   language = {eng},
   issn = {0016-6731},
   journal = {Genetics},
   title = {Analysis of rad-51 separation of function allele suggests divergence of the synthesis-dependent strand annealing and double Holliday junction pathways prior to RAD-51 filament disassembly},
   url = {https://academic.oup.com/genetics/article/230/2/iyaf063/8107673?login=true},
   volume = {230},
   year = {2025}
}

TY  - JOUR
ID  - 2511225
AU  - Oberlitner, Joseph - Tinman, Maggie - Das, Aasthika - Koury, Emily - Silva, Nicola - Smolikove, Sarit
PY  - 2025
TI  - Analysis of rad-51 separation of function allele suggests divergence of the synthesis-dependent strand annealing and double Holliday junction pathways prior to RAD-51 filament disassembly
JF  - Genetics
VL  - 230
IS  - 2
SP  - 1-16
EP  - 1-16
PB  - OXFORD UNIV PRESS
SN  - 00166731
KW  - C. elegans
KW  - meiosis
KW  - RAD51
KW  - crossover
UR  - https://academic.oup.com/genetics/article/230/2/iyaf063/8107673?login=true
N2  - DNA double-strand breaks (DSBs) are formed in meiosis, so their repair in the homologous recombination (HR) pathway will lead to crossover formation, which is essential for successful chromosome segregation. HR contains 2 subpathways: synthesis-dependent strand annealing (SDSA) that creates noncrossover and double Holliday junction (dHJ) that generates crossovers. RAD-51 is a protein essential to the formation of all products of HR, as it assembles on the processed DSB, allowing the invasion of the single-stranded DNA into a region of homology. RAD-51 is removed by RAD-54.L after invasion to allow for repair to occur. Here, we investigate a separation of function allele of rad-51, rad-51::FLAG, as compared to 2 other RAD-51 alleles: rad-51::degron and GFP::rad-51. rad-51::FLAG displays slowed repair kinetics, resulting in an accumulation of RAD-51 foci. rad-51::FLAG worms also activate the DSB checkpoint, but to a less extant than that of rad-51 null mutants. In a proximity ligation assay, RAD-54.L and RAD-51 show enriched colocalization in rad-51::FLAG germlines (but not in rad-51::degron), consistent with stalling at the strand invasion step in HR. The defects in RAD-51 disassembly in rad-51::FLAG mutants lead to formation of chromosomal fragments, similar in their magnitude to ones observed in rad-51 or rad-54.L null mutants. However, rad-51::FLAG mutants (unlike a rad-51 null, GFP::rad-51 or rad-54.L null mutants) displayed no defects in the formation of crossover-designated sites (via GFP::COSA-1 localization). Given that rad-51::FLAG worms show checkpoint activation and chromosomal fragments, these results suggest that crossover repair concludes normally, while the noncrossover pathway is perturbed. This is strikingly different from rad-51::degron and GFP::rad-51 strains, which are proficient or deficient in both pathways, respectively. These results suggest that noncrossovers vs crossovers have distinct recombination intermediates and diverge prior to RAD-51 disassembly.
ER  -

OBERLITNER, Joseph; Maggie TINMAN; Aasthika DAS; Emily KOURY; Nicola SILVA a Sarit SMOLIKOVE. Analysis of rad-51 separation of function allele suggests divergence of the synthesis-dependent strand annealing and double Holliday junction pathways prior to RAD-51 filament disassembly. \textit{Genetics}. CARY: OXFORD UNIV PRESS, 2025, roč.~230, č.~2, s.~1-16. ISSN~0016-6731. Dostupné z: https://doi.org/10.1093/genetics/iyaf063.

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