{"doi":"10.1101/2022.08.23.504870","title":"CasDinG is an ATP-dependent 5’-3’ DNA helicase with accessory domains essential for type IV CRISPR immunity","abstract":"ABSTRACT CRISPR-associated DinG protein (CasDinG) is essential to type IV-A CRISPR function. However, the enzymatic activities of CasDinG are unknown. Here we demonstrate that CasDinG from Pseudomonas aeruginosa strain 83 is an ATP- and metal-dependent 5’-3’ DNA helicase. The crystal structure of CasDinG reveals a helicase core of two RecA-like domains with three accessory domains (N-terminal, arch, and vestigial FeS). To examine the in vivo function of these CasDinG domains, we first identified the preferred PAM sequence (5’-GNAWN-3’ on the 5’-side of the target) with a plasmid library containing all combinations of the five nucleotides upstream of the target sequence. Plasmid clearance assays (using a 5’-GGAAA-3’ PAM) with CasDinG domain mutants demonstrated the vFeS and arch accessory domains are both essential for type IV immunity. These results provide a needed structural and biochemical framework for understanding the type IV-A CRISPR system.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2022,"id":300369,"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":3,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.962,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":990847,"name":"Christian Cahoon","orcid":"0009-0009-4894-4637","position":1,"is_corresponding":false},{"id":991395,"name":"Matthew J. Armbrust","orcid":null,"position":2,"is_corresponding":false},{"id":991396,"name":"Olive Redman","orcid":null,"position":3,"is_corresponding":false},{"id":990848,"name":"Aaron Thomas","orcid":"0000-0003-3071-084X","position":4,"is_corresponding":false},{"id":567564,"name":"Ryan N. Jackson","orcid":"0000-0001-7403-3794","position":5,"is_corresponding":false},{"id":990846,"name":"Hannah Domgaard","orcid":"0000-0001-9359-8352","position":0,"is_corresponding":true}],"reference_count":49,"raw_metadata":null,"created_at":"2026-07-19T00:31:53.559757Z","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":[]}