@article{5278f420bdc9417abefc1476fb87f5ff,
title = "QTQTN motif upstream of the furin-cleavage site plays a key role in SARS-CoV-2 infection and pathogenesis",
abstract = "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 vitro 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 (ΔQTQTN). Here, we report that the QTQTN deletion attenuates viral replication in respiratory cells in vitro and attenuates disease in vivo. 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 SARSCoV- 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-the FCS, loop length, and glycosylation-are required for efficient SARS-CoV-2 replication and pathogenesis.",
keywords = "QTQTN, SARS-CoV-2, furin cleavage site, glycosylation, spike",
author = "Vu, {Michelle N.} and Lokugamage, {Kumari G.} and Plante, {Jessica A.} and Dionna Scharton and Bailey, {Aaron O.} and Stephanea Sotcheff and Swetnam, {Daniele M.} and Johnson, {Bryan A.} and Craig Schindewolf and {Elias Alvarado}, R. and Crocquet-Valdes, {Patricia A.} and Kari Debbink and Weaver, {Scott C.} and Walker, {David H.} and Russell, {William K.} and Routh, {Andrew L.} and Plante, {Kenneth S.} and Menachery, {Vineet D.}",
note = "Funding Information: Data Availability. The raw [Transcriptomics] data have been deposited with links to NCBI BioProject databased under BioProject ID PRJNA856125 (32). The mass spectrometry proteomics data have been deposited to the ProteomeX-change Consortium via the PRIDE partner repository with the dataset identifier PXD034332 (33) and 10.6019/PXD034332 (34) ACKNOWLEDGMENTS. Research was supported by grants from National Institute of Allergy and Infectious Diseases (NIAID) of the NIH (R01-AI153602 and R21-AI145400 to V.D.M.; R24-AI120942 [WRCEVA] to S.C.W.). W.K.R. was supported by the Cancer Prevention Research Institute of Texas (RP190682). A.L.R. was supported by an Institute of Human Infection and Immunity at UTMB COVID-19 Research Fund. Research was also supported by STARs Award provided by the University of Texas System to V.D.M. Trainee funding provided by NIAID of the NIH to M.N.V. (T32-AI060549). We thank Shinji Makino for gifting the nucleocapsid antibody. Figures were created with BioRender.com. Publisher Copyright: {\textcopyright} 2022 National Academy of Sciences. All rights reserved.",
year = "2022",
month = aug,
day = "9",
doi = "10.1073/pnas.2205690119",
language = "English (US)",
volume = "119",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "National Academy of Sciences",
number = "32",
}