{"doi":"10.1101/2024.06.14.599120","title":"Rad51 determines pathway usage in post-replication repair","abstract":"ABSTRACT Stalled replication forks can be processed by several distinct mechanisms collectively called post-replication repair which includes homologous recombination, fork regression, and translesion DNA synthesis. However, the regulation of the usage between these pathways is not fully understood. The Rad51 protein plays a pivotal role in maintaining genomic stability through its roles in HR and in protecting stalled replication forks from degradation. We report the isolation of separation-of-function mutations in Saccharomyces cerevisiae Rad51 that retain their recombination function but display a defect in fork protection leading to a shift in post-replication repair pathway usage from HR to alternate pathways including mutagenic translesion synthesis. Rad51-E135D and Rad51-K305N show normal in vivo and in vitro recombination despite changes in their DNA binding profiles, in particular to dsDNA, with a resulting effect on their ATPase activities. The mutants lead to a defect in Rad51 recruitment to stalled forks in vivo as well as a defect in the protection of dsDNA from degradation by Dna2-Sgs1 and Exo1 in vitro . A high-resolution cryo-electron microscopy structure of the Rad51-ssDNA filament at 2.4 Å resolution provides a structural basis for a mechanistic understanding of the mutant phenotypes. Together, the evidence suggests a model in which Rad51 binding to duplex DNA is critical to control pathway usage at stalled replication forks.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2024,"id":488623,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":2,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9516,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1333587,"name":"Shannon J. Ceballos","orcid":null,"position":1,"is_corresponding":false},{"id":1333588,"name":"Steven K Gore","orcid":null,"position":2,"is_corresponding":false},{"id":478808,"name":"Jie Liu","orcid":"0000-0001-5817-9144","position":3,"is_corresponding":false},{"id":824081,"name":"Giordano Reginato","orcid":"0009-0002-5284-4339","position":4,"is_corresponding":false},{"id":1333589,"name":"María I. Cano-Linares","orcid":null,"position":5,"is_corresponding":false},{"id":1333232,"name":"Katarzyna H. Masłowska","orcid":"0000-0003-3393-8225","position":6,"is_corresponding":false},{"id":1333233,"name":"Florencia Villafañez","orcid":"0000-0001-9835-2343","position":7,"is_corresponding":false},{"id":1333590,"name":"Christopher Ede","orcid":null,"position":8,"is_corresponding":false},{"id":562234,"name":"Vincent Pagès","orcid":"0000-0002-3059-2903","position":9,"is_corresponding":false},{"id":1333234,"name":"Félix Prado","orcid":"0000-0001-9805-782X","position":10,"is_corresponding":false},{"id":316515,"name":"Petr Ćejka","orcid":"0000-0002-9087-032X","position":11,"is_corresponding":false},{"id":168557,"name":"Wolf-Dietrich Heyer","orcid":"0000-0002-7774-1953","position":12,"is_corresponding":false},{"id":542276,"name":"Damon Meyer","orcid":"0000-0002-1889-4335","position":0,"is_corresponding":true}],"reference_count":108,"raw_metadata":null,"created_at":"2026-07-19T02:08:19.655720Z","pmid":"38915629","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}