{"doi":"10.1016/j.cell.2024.03.022","title":"Positive selection CRISPR screens reveal a druggable pocket in an oligosaccharyltransferase required for inflammatory signaling to NF-κB","abstract":"Nuclear factor κB (NF-κB) plays roles in various diseases. Many inflammatory signals, such as circulating lipopolysaccharides (LPSs), activate NF-κB via specific receptors. Using whole-genome CRISPR-Cas9 screens of LPS-treated cells that express an NF-κB-driven suicide gene, we discovered that the LPS receptor Toll-like receptor 4 (TLR4) is specifically dependent on the oligosaccharyltransferase complex OST-A for N -glycosylation and cell-surface localization. The tool compound NGI-1 inhibits OST complexes in vivo , but the underlying molecular mechanism remained unknown. We did a CRISPR base-editor screen for NGI-1-resistant variants of STT3A, the catalytic subunit of OST-A. These variants, in conjunction with cryoelectron microscopy studies, revealed that NGI-1 binds the catalytic site of STT3A, where it traps a molecule of the donor substrate dolichyl-PP-GlcNAc 2 -Man 9 -Glc 3 , suggesting an uncompetitive inhibition mechanism. Our results provide a rationale for and an initial step toward the development of STT3A-specific inhibitors and illustrate the power of contemporaneous base-editor and structural studies to define drug mechanism of action.","journal":"Cell","year":2024,"id":418623,"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":48,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9464,"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":1021334,"name":"Ana S. Ramίrez","orcid":"0000-0002-1619-0721","position":1,"is_corresponding":false},{"id":623097,"name":"Marta Baro","orcid":"0000-0002-6019-0534","position":2,"is_corresponding":false},{"id":704444,"name":"Lixia He","orcid":null,"position":3,"is_corresponding":false},{"id":80464,"name":"Mudra Hegde","orcid":"0000-0003-4530-4633","position":4,"is_corresponding":false},{"id":679649,"name":"Vidyasagar Koduri","orcid":"0000-0002-9700-8169","position":5,"is_corresponding":false},{"id":1207810,"name":"Jamie L. Pfaff","orcid":null,"position":6,"is_corresponding":false},{"id":110476,"name":"Ruth E. Hanna","orcid":"0000-0001-5586-2300","position":7,"is_corresponding":false},{"id":632978,"name":"Julia Kowal","orcid":"0000-0003-4694-1896","position":8,"is_corresponding":false},{"id":1049701,"name":"Nitin H. Shirole","orcid":"0000-0002-2579-456X","position":9,"is_corresponding":false},{"id":785065,"name":"Yanfeng He","orcid":"0000-0003-1707-2353","position":10,"is_corresponding":false},{"id":54229,"name":"John G. Doench","orcid":"0000-0002-3707-9889","position":11,"is_corresponding":false},{"id":413470,"name":"Joseph N. Contessa","orcid":"0000-0001-9408-1865","position":12,"is_corresponding":false},{"id":292313,"name":"Kaspar P. Locher","orcid":"0000-0002-8207-2889","position":13,"is_corresponding":false},{"id":63522,"name":"William G. Kaelin","orcid":"0000-0002-0574-4856","position":14,"is_corresponding":false},{"id":636325,"name":"Benjamin L. Lampson","orcid":"0000-0002-6033-3363","position":0,"is_corresponding":true}],"reference_count":81,"raw_metadata":null,"created_at":"2026-07-19T01:57:09.067930Z","pmid":"38670073","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":[]}