{"doi":"10.1093/nar/gkae256","title":"Assembly of SARS-CoV-2 nucleocapsid protein with nucleic acid","abstract":"The viral genome of SARS-CoV-2 is packaged by the nucleocapsid (N-)protein into ribonucleoprotein particles (RNPs), 38 ± 10 of which are contained in each virion. Their architecture has remained unclear due to the pleomorphism of RNPs, the high flexibility of N-protein intrinsically disordered regions, and highly multivalent interactions between viral RNA and N-protein binding sites in both N-terminal (NTD) and C-terminal domain (CTD). Here we explore critical interaction motifs of RNPs by applying a combination of biophysical techniques to ancestral and mutant proteins binding different nucleic acids in an in vitro assay for RNP formation, and by examining nucleocapsid protein variants in a viral assembly assay. We find that nucleic acid-bound N-protein dimers oligomerize via a recently described protein-protein interface presented by a transient helix in its long disordered linker region between NTD and CTD. The resulting hexameric complexes are stabilized by multivalent protein-nucleic acid interactions that establish crosslinks between dimeric subunits. Assemblies are stabilized by the dimeric CTD of N-protein offering more than one binding site for stem-loop RNA. Our study suggests a model for RNP assembly where N-protein scaffolding at high density on viral RNA is followed by cooperative multimerization through protein-protein interactions in the disordered linker.","journal":"Nucleic Acids Research","year":2024,"id":418982,"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":44,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9549,"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":2371,"name":"Abdullah M. Syed","orcid":"0000-0001-9156-9662","position":1,"is_corresponding":false},{"id":568701,"name":"Mir M. Khalid","orcid":"0000-0002-1862-6795","position":2,"is_corresponding":false},{"id":398409,"name":"Ai Nguyen","orcid":null,"position":3,"is_corresponding":false},{"id":2372,"name":"Alison Ciling","orcid":"0000-0002-8393-6829","position":4,"is_corresponding":false},{"id":313756,"name":"Di Wu","orcid":"0000-0002-3440-1860","position":5,"is_corresponding":false},{"id":337240,"name":"Wai‐Ming Yau","orcid":"0000-0002-7668-7780","position":6,"is_corresponding":false},{"id":261725,"name":"Sanjana Srinivasan","orcid":"0000-0002-1439-1052","position":7,"is_corresponding":false},{"id":267809,"name":"Dominic Esposito","orcid":"0000-0002-9987-1687","position":8,"is_corresponding":false},{"id":2407,"name":"Jennifer A. Doudna","orcid":"0000-0001-9161-999X","position":9,"is_corresponding":false},{"id":313757,"name":"Grzegorz Piszczek","orcid":"0000-0002-5270-3678","position":10,"is_corresponding":false},{"id":103701,"name":"Mélanie Ott","orcid":"0000-0002-5697-1274","position":11,"is_corresponding":false},{"id":396759,"name":"Peter Schuck","orcid":"0000-0002-8859-6966","position":12,"is_corresponding":false},{"id":396757,"name":"Huaying Zhao","orcid":"0000-0002-8827-6639","position":0,"is_corresponding":true}],"reference_count":66,"raw_metadata":null,"created_at":"2026-07-19T01:57:09.067930Z","pmid":"38587193","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":[]}