{"doi":"10.1101/2024.03.28.587234","title":"Wrapping of single-stranded DNA by Replication Protein A and modulation through phosphorylation","abstract":"Single-stranded DNA (ssDNA) intermediates, which emerge during DNA metabolic processes are shielded by Replication Protein A (RPA). RPA binds to ssDNA and acts as a gatekeeper, directing the ssDNA towards downstream DNA metabolic pathways with exceptional specificity. Understanding the mechanistic basis for such RPA-dependent specificity requires a comprehensive understanding of the structural conformation of ssDNA when bound to RPA. Previous studies suggested a stretching of ssDNA by RPA. However, structural investigations uncovered a partial wrapping of ssDNA around RPA. Therefore, to reconcile the models, in this study, we measured the end-to-end distances of free ssDNA and RPA-ssDNA complexes using single-molecule FRET and Double Electron-Electron Resonance (DEER) spectroscopy and found only a small systematic increase in the end-to-end distance of ssDNA upon RPA binding. This change does not align with a linear stretching model but rather supports partial wrapping of ssDNA around the contour of DNA binding domains of RPA. Furthermore, we reveal how phosphorylation at the key Ser-384 site in the RPA70 subunit provides access to the wrapped ssDNA by remodeling the DNA-binding domains. These findings establish a precise structural model for RPA-bound ssDNA, providing valuable insights into how RPA facilitates the remodeling of ssDNA for subsequent downstream processes.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2024,"id":496107,"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":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9565,"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":911093,"name":"Vikas Kaushik","orcid":"0000-0001-6629-7693","position":1,"is_corresponding":false},{"id":1229888,"name":"Iram Munir Ahmad","orcid":null,"position":2,"is_corresponding":false},{"id":477330,"name":"Jaigeeth Deveryshetty","orcid":"0000-0003-1979-4085","position":3,"is_corresponding":false},{"id":107330,"name":"Alex S. Holehouse","orcid":"0000-0002-4155-5729","position":4,"is_corresponding":false},{"id":1343017,"name":"Snorri Th.d Sigurdsson","orcid":null,"position":5,"is_corresponding":false},{"id":316553,"name":"Brian Bothner","orcid":"0000-0003-1295-9609","position":6,"is_corresponding":false},{"id":890183,"name":"Reza Dastvan","orcid":"0000-0002-4779-275X","position":7,"is_corresponding":false},{"id":452632,"name":"Sofia Origanti","orcid":"0000-0002-6953-0160","position":8,"is_corresponding":false},{"id":452634,"name":"Edwin Antony","orcid":"0000-0002-1888-054X","position":9,"is_corresponding":false},{"id":576227,"name":"Rahul Chadda","orcid":"0000-0001-6710-3985","position":0,"is_corresponding":true}],"reference_count":42,"raw_metadata":null,"created_at":"2026-07-19T02:09:23.333150Z","pmid":"38585962","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":[]}