@article{f6f5766c49a94841a73ab85842e081e3,
title = "The D614G mutation redirects SARS-CoV-2 spike to lysosomes and suppresses deleterious traits of the furin cleavage site insertion mutation",
abstract = "Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) egress occurs by lysosomal exocytosis. We show that the Spike D614G mutation enhances Spike trafficking to lysosomes, drives Spike-mediated reprogramming of lysosomes, and reduces cell surface Spike expression by ~3-fold. D614G is not a human-specific adaptation. Rather, it is an adaptation to the earlier furin cleavage site insertion (FCSI) mutation that occurred at the genesis of SARS-CoV-2. While advantageous to the virus, furin cleavage of spike has deleterious effects on spike structure and function, inhibiting its trafficking to lysosomes and impairing its infectivity by the transmembrane serine protease 2(TMPRSS2)-independent, endolysosomal pathway. D614G restores spike trafficking to lysosomes and enhances the earliest events in SARS-CoV-2 infectivity, while spike mutations that restore SARS-CoV-2{\textquoteright}s TMPRSS2-independent infectivity restore spike{\textquoteright}s trafficking to lysosomes. Together, these and other results show that D614G is an intragenic suppressor of deleterious traits linked to the FCSI and lend additional support to the endolysosomal model of SARS-CoV-2 egress and entry.",
author = "Chenxu Guo and Tsai, {Shang Jui} and Yiwei Ai and Maggie Li and Eduardo Anaya and Andrew Pekosz and Andrea Cox and Gould, {Stephen J.}",
note = "Funding Information: Acknowledgments:W ethankC.MachamerandF .Hochbergforinsightsduringthecourseof thisstudy,L.A.RokerandB.SmithoftheJohnsHopkinsSchoolofMedicineMicroscopeFacility forassistancewithconfocalimageacquisitionandelectronmicroscopy,J.Morrellforassistance withplasmidassembly,G.Massaccesiforassistanceinplasmasamplehandling,andJ.Franklin andtheJHUsynthesisandsequencingfacilityforDNAsequenceanalysis.W ealsothankthe NationalInstituteofInfectiousDiseases,Japan,forpro viding VeroE6TMPRSS2cells.The specimensusedforthispublicationwerepartoftheJohnsHopkinsBiospecimenRepository, whichisbased on thecontributionof many patients, research teams,and clinicians. Funding: ThisworkwassupportedbygrantsfromtheNationalInstitutesofHealth(NIH)NationalCancer Institute(UG3CA241687andR35HL150807)toS.J.G.,grantfromtheNIHCenterofExcellencein InfluenzaResearchandSurveillance(HHSN272201400007C)toA.P ., andgrantfromtheNIH/ NationalInstituteofAllergyandInfectiousDiseases(U19A1088791)toA.C.Thisworkwasalso supported bya sponsored research agreement from Capricor Inc. to Johns Hopkins University (S.J.G.)andbyfundsfromAstraZenecaandJohnsHopkinsUniversity-Astr aZeneca Scholarship programtoC.G.,aJohnsHopkinsUniversity-AstraZenecaScholar.Authorcontributions: Conceptualization:C.G.,S.-J.T ., A.P ., andS.J.G.Methodology:C.G.,S.-J.T ., E.A.,A.P ., A.C.,andS.J.G. Investiga tion: C.G.,Y .A., M.L.,E.A.,A.P ., andS.J.G.Visualization:C.G.,Y .A., andS.J.G.Supervision: A.P ., A.C.,andS.J.G.Writing(originaldraft):C.G.andS.J.G.Writing(reviewandediting):C.G.,S.-J.T ., Y .A., M.L.,E.A.,A.P ., A.C.,andS.J.G.Competinginterests:S.J.G.,C.G.,andS.-J.T .are beneficiariesofintellectualpropertiesheldbyJohnsHopkinsUniversity,someofwhichare relevant to this paper. Capricor has licensed intellectual property from Johns Hopkins University that is related to these studies, and S.J.G. is a paid consultant for Capricor, holds equityinCapricor,andiscoinventorofintellectualpropertylicensedbyCapricor.C.G.,S.-J.T ., andS.J.G.arecoinventorsofintellectualpropertylicensedbyCapricor.Theauthorsdeclarethat theyhavenoothercompetinginterests.Dataandmaterialsavailability:Alldataneededto evaluatetheconclusionsinthepaperarepresentinthepaperand/ortheSupplementary Materials. Funding Information: We thank C. Machamer and F. Hochberg for insights during the course of this study, L. A. Roker and B. Smith of the Johns Hopkins School of Medicine Microscope Facility for assistance with confocal image acquisition and electron microscopy, J. Morrell for assistance with plasmid assembly, G. Massaccesi for assistance in plasma sample handling, and J. Franklin and the JHU synthesis and sequencing facility for DNA sequence analysis. We also thank the National Institute of Infectious Diseases, Japan, for providing VeroE6TMPRSS2 cells. The specimens used for this publication were part of the Johns Hopkins Biospecimen Repository, which is based on the contribution of many patients, research teams, and clinicians. This work was supported by grants from the National Institutes of Health (NIH) National Cancer Institute (UG3CA241687 and R35HL150807) to S.J.G., grant from the NIH Center of Excellence in Influenza Research and Surveillance (HHSN272201400007C) to A.P., and grant from the NIH/ National Institute of Allergy and Infectious Diseases (U19A1088791) to A.C. This work was also supported by a sponsored research agreement from Capricor Inc. to Johns Hopkins University (S.J.G.) and by funds from AstraZeneca and Johns Hopkins University-AstraZeneca Scholarship program to C.G., a Johns Hopkins University-AstraZeneca Scholar. Publisher Copyright: Copyright {\textcopyright} 2022 The Authors, some rights reserved.",
year = "2022",
month = dec,
doi = "10.1126/sciadv.ade5085",
language = "English (US)",
volume = "8",
journal = "Science Advances",
issn = "2375-2548",
publisher = "American Association for the Advancement of Science",
number = "51",
}