{"doi":"10.1101/2023.02.05.527205","title":"Homodecameric Rad52 promotes single-position Rad51 nucleation in homologous recombination","abstract":"Abstract Homologous recombination (HR) is a pathway for the accurate repair of double-stranded DNA breaks. These breaks are resected to yield single-stranded DNA (ssDNA) that are coated by Replication Protein A (RPA). Saccharomyces cerevisiae Rad52 is a mediator protein that promotes HR by facilitating formation of Rad51 nucleoprotein filaments on RPA-coated ssDNA. Canonically, Rad52 has been described to function by displacing RPA to promote Rad51 binding. However, in vitro , Rad51 readily forms a filament by displacing RPA in the absence of Rad52. Yet, in vivo , Rad52 is essential for HR. Here, we resolve how Rad52 functions as a mediator using single-particle cryo-electron microscopy and biophysical approaches. We show that Rad52 functions as a homodecamer and catalyzes single-position nucleation of Rad51. The N-terminal half of Rad52 is a well-ordered ring, while the C-terminal half is disordered. An intrinsic asymmetry within Rad52 is observed, where one or a few of the C-terminal halves interact with the ordered N-terminal ring. Within the C-terminal half, we identify two conserved charged patches that harbor the Rad51 and RPA interacting motifs. Interactions between these two charged patches regulate a ssDNA binding. These features drive Rad51 binding to a single position on the Rad52 decameric ring. We propose a Rad52 catalyzed single-position nucleation model for the formation of pre-synaptic Rad51 filaments in HR.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2023,"id":391653,"datarank":0.29188652235829704,"base_score":1.9459101490553132,"endowment":1.9459101490553132,"self_citation_contribution":0.29188652235829704,"citation_network_contribution":0.0,"self_endowment_contribution":0.29188652235829704,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":6,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9558,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":576227,"name":"Rahul Chadda","orcid":"0000-0001-6710-3985","position":1,"is_corresponding":false},{"id":453767,"name":"Jenna R. Mattice","orcid":null,"position":2,"is_corresponding":false},{"id":1050066,"name":"Simrithaa Karunakaran","orcid":null,"position":3,"is_corresponding":false},{"id":585484,"name":"Michael Rau","orcid":"0000-0001-5170-2000","position":4,"is_corresponding":false},{"id":272060,"name":"Katherine Basore","orcid":"0000-0001-5974-0968","position":5,"is_corresponding":false},{"id":477331,"name":"Nilisha Pokhrel","orcid":"0000-0002-2957-4046","position":6,"is_corresponding":false},{"id":1050067,"name":"Noah Englander","orcid":null,"position":7,"is_corresponding":false},{"id":420916,"name":"James A. J. Fitzpatrick","orcid":"0000-0002-7907-3602","position":8,"is_corresponding":false},{"id":316553,"name":"Brian Bothner","orcid":"0000-0003-1295-9609","position":9,"is_corresponding":false},{"id":452634,"name":"Edwin Antony","orcid":"0000-0002-1888-054X","position":10,"is_corresponding":false},{"id":477330,"name":"Jaigeeth Deveryshetty","orcid":"0000-0003-1979-4085","position":0,"is_corresponding":true}],"reference_count":77,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T01:18:47.408273Z","pmid":"36778491","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":[]}