{"doi":"10.1101/2023.01.18.524314","title":"The BR-body proteome contains a complex network of protein-protein and protein-RNA interactions","abstract":"Abstract Bacterial RNP bodies (BR-bodies) are non-membrane-bound structures that facilitate mRNA decay by concentrating mRNA substrates with RNase E and the associated RNA degradosome machinery. However, the full complement of proteins enriched in BR-bodies has not been defined. Here we define the protein components of BR-bodies through enrichment of the bodies followed by mass spectrometry-based proteomic analysis. We found 111 BR-body enriched proteins, including several RNA binding proteins, many of which are also recruited directly to in vitro reconstituted RNase E droplets, showing BR-bodies are more complex than previously assumed. While most BR-body enriched proteins that were tested cannot phase separate, we identified five that undergo RNA-dependent phase separation in vitro , showing other RNP condensates interface with BR-bodies. RNA degradosome protein clients are recruited more strongly to RNase E droplets than droplets of other RNP condensates, implying that client specificity is largely achieved through direct protein-protein interactions. We observe that some RNP condensates assemble with preferred directionally, suggesting that RNA may be trafficked through RNP condensates in an ordered manner to facilitate mRNA processing/decay, and that some BR-body associated proteins have the capacity to dissolve the condensate. Finally, we find that RNA dramatically stimulates the rate of RNase E phase separation in vitro , explaining the dissolution of BR-bodies after cellular mRNA depletion observed previously. Altogether, these results suggest that a complex network of protein-protein and protein-RNA interactions controls BR-body phase separation and RNA processing. Highlights BR-body proteomics identified 111 proteins enriched in BR-bodies. BR-bodies associate with an interconnected network of RNP condensates. BR-body condensation is modulated by its interaction network. RNA is required for rapid BR-body condensation. Graphical Abstract Summary of the BR-body protein interactome. Lines between two protein circles represent a direct interaction.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2023,"id":395667,"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":0.9467,"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":1045247,"name":"Imalka W. Rathnayaka-Mudiyanselage","orcid":"0000-0002-8636-2525","position":1,"is_corresponding":false},{"id":274189,"name":"Nisansala S. Muthunayake","orcid":"0000-0003-4916-7453","position":2,"is_corresponding":false},{"id":1045248,"name":"Ali Hatami","orcid":"0000-0002-5891-4900","position":3,"is_corresponding":false},{"id":1171268,"name":"Mousseau C.B.","orcid":null,"position":4,"is_corresponding":false},{"id":1171269,"name":"Ortiz-Rodríguez L.A.","orcid":null,"position":5,"is_corresponding":false},{"id":1045871,"name":"Jamuna Vaishnav","orcid":null,"position":6,"is_corresponding":false},{"id":1045249,"name":"Michael Collins","orcid":"0000-0002-0272-9968","position":7,"is_corresponding":false},{"id":1171270,"name":"Arjet Gega","orcid":null,"position":8,"is_corresponding":false},{"id":1171271,"name":"M. S","orcid":null,"position":9,"is_corresponding":false},{"id":347384,"name":"Hadi M. Yassine","orcid":"0000-0001-7592-2788","position":10,"is_corresponding":false},{"id":731234,"name":"Agnidipta Ghosh","orcid":"0000-0002-7753-0240","position":11,"is_corresponding":false},{"id":242841,"name":"Julie S. Biteen","orcid":"0000-0003-2038-6484","position":12,"is_corresponding":false},{"id":1170904,"name":"Ye Zhu","orcid":"0000-0002-7086-0925","position":13,"is_corresponding":false},{"id":1171272,"name":"Cline Marcella M.","orcid":null,"position":14,"is_corresponding":false},{"id":1171273,"name":"Childers W.S.","orcid":null,"position":15,"is_corresponding":false},{"id":1171274,"name":"Schrader J.M.","orcid":null,"position":16,"is_corresponding":false},{"id":657237,"name":"Vidhyadhar Nandana","orcid":"0000-0003-0564-8343","position":0,"is_corresponding":true}],"reference_count":75,"raw_metadata":null,"created_at":"2026-07-19T01:19:22.857556Z","pmid":"36712072","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":[]}