{"doi":"10.1093/nar/gkab284","title":"<i>E. coli</i> RNase I exhibits a strong Ca2+-dependent inherent double-stranded RNase activity","abstract":"<jats:title>Abstract</jats:title>\n               <jats:p>Since its initial characterization, Escherichia coli RNase I has been described as a single-strand specific RNA endonuclease that cleaves its substrate in a largely sequence independent manner. Here, we describe a strong calcium (Ca2+)-dependent activity of RNase I on double-stranded RNA (dsRNA), and a Ca2+-dependent novel hybridase activity, digesting the RNA strand in a DNA:RNA hybrid. Surprisingly, Ca2+ does not affect the activity of RNase I on single stranded RNA (ssRNA), suggesting a specific role for Ca2+ in the modulation of RNase I activity. Mutation of a previously overlooked Ca2+ binding site on RNase I resulted in a gain-of-function enzyme that is highly active on dsRNA and could no longer be stimulated by the metal. In summary, our data imply that native RNase I contains a bound Ca2+, allowing it to target both single- and double-stranded RNAs, thus having a broader substrate specificity than originally proposed for this traditional enzyme. In addition, the finding that the dsRNase activity, and not the ssRNase activity, is associated with the Ca2+-dependency of RNase I may be useful as a tool in applied molecular biology.</jats:p>","journal":"Nucleic Acids Research","year":2021,"id":16483,"datarank":0.8665702328202206,"base_score":2.995732273553991,"endowment":2.995732273553991,"self_citation_contribution":0.4493598410330987,"citation_network_contribution":0.41721039178712194,"self_endowment_contribution":0.4493598410330987,"citer_contribution":0.41721039178712194,"corpus_percentile":null,"corpus_rank":null,"citation_count":19,"citer_count":18,"citers_with_citation_signal":15,"citers_with_endowment":15,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":121500,"name":"Baptiste Coxam","orcid":null,"position":1,"is_corresponding":false},{"id":121501,"name":"Tien-Hao Chen","orcid":null,"position":2,"is_corresponding":false},{"id":121502,"name":"Nan Dai","orcid":null,"position":3,"is_corresponding":false},{"id":121503,"name":"Lana Saleh","orcid":null,"position":4,"is_corresponding":false},{"id":121504,"name":"Ivan R Corrêa","orcid":"0000-0002-3169-6878","position":5,"is_corresponding":false},{"id":121505,"name":"Nicole M Nichols","orcid":null,"position":6,"is_corresponding":false},{"id":121506,"name":"Erbay Yigit","orcid":"0000-0003-2637-4994","position":7,"is_corresponding":false},{"id":121499,"name":"Sebastian Grünberg","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":2.995732273553991,"endowment":2.995732273553991,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"33885787","pmcid":"PMC8136782","openalex_id":"https://openalex.org/W3153561079","authors":[],"funders":[{"funder_name":"New England Biolabs","grant_id":"","title":null}],"total_grants":1,"fwci":1.2212,"citation_percentile":0.78417537,"influential_citations":1,"citation_trend":[{"year":2021,"count":2},{"year":2022,"count":3},{"year":2023,"count":5},{"year":2024,"count":5},{"year":2025,"count":3},{"year":2026,"count":1}],"oa_status":"gold","license":"cc-by-nc","oa_locations":[{"url":"https://academic.oup.com/nar/article-pdf/49/9/5265/38001519/gkab284.pdf","host_type":"journal"},{"url":"https://academic.oup.com/nar/article-pdf/49/9/5265/38001519/gkab284.pdf","host_type":"GOLD"},{"url":"https://academic.oup.com/nar/article-pdf/49/9/5265/38001519/gkab284.pdf","host_type":"publisher"},{"url":"http://academic.oup.com/nar/article-pdf/49/9/5265/38001519/gkab284.pdf","host_type":"publisher"},{"url":"https://doi.org/10.1093/nar/gkab284","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/33885787","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/8136782","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC8136782","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC8136782?pdf=render","host_type":"Europe_PMC"}],"fields_of_study":["RNA and protein synthesis mechanisms","Bacteriophages and microbial interactions","Bacterial Genetics and Biotechnology","Biology","Medicine","Chemistry","Amino Acid Substitution","Calcium","DNA","Endoribonucleases","Metals","RNA","RNA, Double-Stranded","Ribonucleases","Substrate Specificity"],"mesh_terms":["Calcium","DNA","Endoribonucleases","Metals","Ribonucleases","RNA","RNA, Double-Stranded","Substrate Specificity","Amino Acid Substitution"],"keywords":["RNase P","RNase MRP","Biology","RNase H","RNA","RNase PH","Endonuclease","Ribonuclease III","Degradosome","RNA silencing","Molecular biology","Biochemistry","DNA","RNA interference","Gene"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-06-02T09:25:32.890674Z","pmid":null,"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":[]}