{"doi":"10.1101/2023.04.18.537104","title":"A single inactivating amino acid change in the SARS-CoV-2 NSP3 Mac1 domain attenuates viral replication and pathogenesis <i>in vivo</i>","abstract":"Abstract Despite unprecedented efforts, our therapeutic arsenal against SARS-CoV-2 remains limited. The conserved macrodomain 1 (Mac1) in NSP3 is an enzyme exhibiting ADP-ribosylhydrolase activity and a possible drug target. To determine the therapeutic potential of Mac1 inhibition, we generated recombinant viruses and replicons encoding a catalytically inactive NSP3 Mac1 domain by mutating a critical asparagine in the active site. While substitution to alanine (N40A) reduced catalytic activity by ∼10-fold, mutations to aspartic acid (N40D) reduced activity by ∼100-fold relative to wildtype. Importantly, the N40A mutation rendered Mac1 unstable in vitro and lowered expression levels in bacterial and mammalian cells. When incorporated into SARS-CoV-2 molecular clones, the N40D mutant only modestly affected viral fitness in immortalized cell lines, but reduced viral replication in human airway organoids by 10-fold. In mice, N40D replicated at &gt;1000-fold lower levels compared to the wildtype virus while inducing a robust interferon response; all animals infected with the mutant virus survived infection and showed no signs of lung pathology. Our data validate the SARS-CoV-2 NSP3 Mac1 domain as a critical viral pathogenesis factor and a promising target to develop antivirals.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2023,"id":394449,"datarank":0.2550781624460599,"base_score":1.3862943611198906,"endowment":1.3862943611198906,"self_citation_contribution":0.20794415416798362,"citation_network_contribution":0.047134008278076275,"self_endowment_contribution":0.20794415416798362,"citer_contribution":0.047134008278076275,"corpus_percentile":null,"corpus_rank":null,"citation_count":3,"citer_count":2,"citers_with_citation_signal":1,"citers_with_endowment":1,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9598,"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":286245,"name":"Rahul K. Suryawanshi","orcid":"0000-0001-8374-669X","position":1,"is_corresponding":false},{"id":562758,"name":"Irene P. Chen","orcid":"0000-0002-5766-9253","position":2,"is_corresponding":false},{"id":552145,"name":"G.J. Correy","orcid":"0000-0001-5155-7325","position":3,"is_corresponding":false},{"id":842015,"name":"Patrick C. O’Leary","orcid":"0000-0002-2919-5943","position":4,"is_corresponding":false},{"id":739776,"name":"Manasi P. Jogalekar","orcid":"0000-0003-1307-4829","position":5,"is_corresponding":false},{"id":931762,"name":"Maria McCavitt-Malvido","orcid":"0009-0006-4731-6047","position":6,"is_corresponding":false},{"id":842014,"name":"Morgan E. Diolaiti","orcid":"0000-0001-5900-3060","position":7,"is_corresponding":false},{"id":892691,"name":"Gabriella R. Kimmerly","orcid":"0000-0002-2920-2218","position":8,"is_corresponding":false},{"id":550948,"name":"Chia-Lin Tsou","orcid":"0000-0002-1205-2970","position":9,"is_corresponding":false},{"id":2423,"name":"Luis Martínez‐Sobrido","orcid":"0000-0001-7084-0804","position":10,"is_corresponding":false},{"id":1363,"name":"Nevan J. Krogan","orcid":"0000-0003-4902-337X","position":11,"is_corresponding":false},{"id":7895,"name":"Alan Ashworth","orcid":"0000-0003-1446-7878","position":12,"is_corresponding":false},{"id":63530,"name":"James S. Fraser","orcid":"0000-0002-5080-2859","position":13,"is_corresponding":false},{"id":103701,"name":"Mélanie Ott","orcid":"0000-0002-5697-1274","position":14,"is_corresponding":false},{"id":385627,"name":"Taha Y. Taha","orcid":"0000-0002-7344-7490","position":0,"is_corresponding":true}],"reference_count":70,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T01:19:14.590635Z","pmid":"37131711","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":[]}