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

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14110%2F25%3A00144389" target="_blank" >RIV/00216224:14110/25:00144389 - isvavai.cz</a>

Result on the web

<a href="https://academic.oup.com/genetics/article/230/2/iyaf063/8107673?login=true" target="_blank" >https://academic.oup.com/genetics/article/230/2/iyaf063/8107673?login=true</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1093/genetics/iyaf063" target="_blank" >10.1093/genetics/iyaf063</a>

Result language

angličtina

Original language name

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

Original language description

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.

Czech name

—

Czech description

—

Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10603 - Genetics and heredity (medical genetics to be 3)

Project

<a href="/en/project/GA23-04918S" target="_blank" >GA23-04918S: A Chromatin Modification-mediated Mechanism Illuminates a Novel Pathway for the Establishment of Meiotic Chromosome Synapsis</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2025

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Genetics

ISSN

0016-6731

e-ISSN

1943-2631

Volume of the periodical

230

Issue of the periodical within the volume

2

Country of publishing house

US - UNITED STATES

Number of pages

16

Pages from-to

1-16

UT code for WoS article

001478853700001

EID of the result in the Scopus database

2-s2.0-105008357992