{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T10:27:28Z","timestamp":1789036048373,"version":"build-2803163510"},"reference-count":34,"publisher":"National Academy of Sciences","issue":"32","license":[{"start":{"date-parts":[[2022,7,26]],"date-time":"2022-07-26T00:00:00Z","timestamp":1658793600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2022,8,9]]},"abstract":"<jats:p>The furin cleavage site (FCS), an unusual feature in the SARS-CoV-2 spike protein, has been spotlighted as a factor key to facilitating infection and pathogenesis by increasing spike processing. Similarly, the QTQTN motif directly upstream of the FCS is also an unusual feature for group 2B coronaviruses (CoVs). The QTQTN deletion has consistently been observed in in\u00a0vitro cultured virus stocks and some clinical isolates. To determine whether the QTQTN motif is critical to SARS-CoV-2 replication and pathogenesis, we generated a mutant deleting the QTQTN motif (\u0394QTQTN). Here, we report that the QTQTN deletion attenuates viral replication in respiratory cells in\u00a0vitro and attenuates disease in\u00a0vivo. The deletion results in a shortened, more rigid peptide loop that contains the FCS and is less accessible to host proteases, such as TMPRSS2. Thus, the deletion reduced the efficiency of spike processing and attenuates SARS-CoV-2 infection. Importantly, the QTQTN motif also contains residues that are glycosylated, and disruption of its glycosylation also attenuates virus replication in a TMPRSS2-dependent manner. Together, our results reveal that three aspects of the S1\/S2 cleavage site\u2014the FCS, loop length, and glycosylation\u2014are required for efficient SARS-CoV-2 replication and pathogenesis.<\/jats:p>","DOI":"10.1073\/pnas.2205690119","type":"journal-article","created":{"date-parts":[[2022,7,26]],"date-time":"2022-07-26T14:37:21Z","timestamp":1658846241000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":45,"title":["QTQTN motif upstream of the furin-cleavage site plays a key role in SARS-CoV-2 infection and pathogenesis"],"prefix":"10.1073","volume":"119","author":[{"given":"Michelle N.","family":"Vu","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kumari G.","family":"Lokugamage","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4768-7458","authenticated-orcid":false,"given":"Jessica A.","family":"Plante","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555"},{"name":"Institute for Human Infection and Immunity, University of Texas Medical Branch, Galveston, TX 77555"},{"name":"World Reference Center of Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, TX 77555"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dionna","family":"Scharton","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555"},{"name":"Institute for Human Infection and Immunity, University of Texas Medical Branch, Galveston, TX 77555"},{"name":"World Reference Center of Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, TX 77555"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8063-3032","authenticated-orcid":false,"given":"Aaron O.","family":"Bailey","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephanea","family":"Sotcheff","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniele M.","family":"Swetnam","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8999-1255","authenticated-orcid":false,"given":"Bryan A.","family":"Johnson","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9804-2246","authenticated-orcid":false,"given":"Craig","family":"Schindewolf","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R. Elias","family":"Alvarado","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555"},{"name":"Institute for Translational Sciences, University of Texas Medical Branch, Galveston, TX"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2079-0076","authenticated-orcid":false,"given":"Patricia A.","family":"Crocquet-Valdes","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7242-7845","authenticated-orcid":false,"given":"Kari","family":"Debbink","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 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Gao SARS-CoV-2 spike protein interacts with multiple innate immune receptors. bioRxiv [Preprint] (2020). https:\/\/doi.org\/10.1101\/2020.07.29.227462.","DOI":"10.1101\/2020.07.29.227462"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2021.08.017"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.02.058"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1009500"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid2606.200516"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00165-13"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2015.06.011"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00818-08"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.03571-13"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkx286"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymeth.2019.01.002"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1101\/gr.229102"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0007960"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1002\/0471250953.bi1320s40"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jproteome.8b00318"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.3389\/fchem.2021.767448"},{"key":"e_1_3_4_32_2","unstructured":"M. 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Deposited 6 June 2022."}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.2205690119","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,7,26]],"date-time":"2022-07-26T14:37:24Z","timestamp":1658846244000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.2205690119"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,26]]},"references-count":34,"journal-issue":{"issue":"32","published-print":{"date-parts":[[2022,8,9]]}},"alternative-id":["10.1073\/pnas.2205690119"],"URL":"https:\/\/doi.org\/10.1073\/pnas.2205690119","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2021.12.15.472450","asserted-by":"object"}]},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,26]]},"assertion":[{"value":"2022-03-31","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-05-26","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-07-26","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e2205690119"}}