{"doi":"10.64898/2025.12.23.696259","title":"Mechanisms of HSV-1 helicase–primase inhibition and replication fork complex assembly","abstract":"Herpesviruses are widespread double-stranded DNA viruses that establish lifelong latency and cause various diseases. Although DNA polymerase-targeting antivirals are effective, increasing drug resistance underscores the need for alternatives. Helicase-primase inhibitors (HPIs) are promising antivirals, but their mechanisms of action are poorly defined. Furthermore, how the helicase-primase (H/P) complex and DNA polymerase coordinate genome replication is not well understood for herpesviruses. Here, we report cryo-EM structures of the herpes simplex virus (HSV) H/P complex bound to HPIs, showing that these lock the helicase-primase complex in an inactive conformation. Single-molecule assays reveal that HPIs cause helicase-primase complexes to pause in unwinding activity on DNA. The structure of an HPI-bound replication fork complex, comprising the H/P complex (UL5, UL52, and UL8) and polymerase holoenzyme (UL30 and UL42), reveals a previously uncharacterized interface bridging these complexes. These findings provide a structural framework for understanding herpesvirus replisome assembly and advancing inhibitor development.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":586936,"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":0.9536,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":521790,"name":"Pradeep Sathyanarayana","orcid":"0000-0002-6195-2430","position":1,"is_corresponding":false},{"id":769448,"name":"Cong Liu","orcid":"0000-0002-1687-2220","position":2,"is_corresponding":false},{"id":1405849,"name":"Pan pan Yang","orcid":null,"position":3,"is_corresponding":false},{"id":146710,"name":"Sandra K. Weller","orcid":"0000-0002-4519-6276","position":4,"is_corresponding":false},{"id":275515,"name":"Mrinal Shekhar","orcid":"0000-0001-8089-8858","position":5,"is_corresponding":false},{"id":339733,"name":"Donald M. Coen","orcid":"0000-0002-2148-5671","position":6,"is_corresponding":false},{"id":471099,"name":"Joseph J. Loparo","orcid":"0000-0003-4941-4696","position":7,"is_corresponding":false},{"id":553557,"name":"Jonathan Abraham","orcid":"0000-0002-7937-3920","position":8,"is_corresponding":false},{"id":1219683,"name":"Zishuo Yu","orcid":"0000-0001-6713-3113","position":0,"is_corresponding":true}],"reference_count":72,"raw_metadata":null,"created_at":"2026-07-19T02:59:32.191237Z","pmid":"41509398","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":[]}