TMEM106B is a receptor mediating ACE2-independent SARS-CoV-2 cell entry

e-pub ahead of print date: 3 July 2023

Published date: 3 August 2023

Additional Information: Funding Information: We thank Niels Willems, Yanicka Smolders, Joni Punjwani, Nathalie van Winkel, Lotte Bral, Jitte Mollekens, and Kristien Minner for performing infections, qPCR, immunofluorescence, cloning, and cell line production and thank Katharina Coun, Jolien Timmermans, and Nathalie Thys for making pseudoviruses. We thank Elisabeth Heylen, Steven Carmans, and Dominique Schols for organizing central lab facilities. We thank John Skehel and David Bauer for suggestions to improve the manuscript; Kai Dallmeier and Johan Neyts for providing hamster anti-SARS-CoV-2 serum; Phil Walker and Andrew Purkiss (Francis Crick Institute) for computer, software, and synchrotron access support; and the staff of beamline I24 of the Diamond Light Source (Oxfordshire, UK) for assistance with data collection. The D.D. laboratory was supported by the Research Foundation – Flanders (FWO; G067423N), Alector LCC, and patent income. The P.C. laboratory and the Structural Biology of Disease Processes Laboratory (lead by Steve Gamblin) were supported by the Francis Crick Institute, funded by Cancer Research UK (FC001061 and FC001078), the UK Medical Research Council (FC001061 and FC001078), and the Wellcome Trust (FC001061 and FC001078). V.C. was supported by a post-doctoral position funded by King's College London, and E.R. was supported by a UKRI Future Leaders fellowship (MR/S015426/1). Cryo-EM data were collected at the London Consortium Electron Microscopy Facility at the Francis Crick Institute funded by the Wellcome Trust (206175/Z/17/Z to Prof. X. Zhang). J.B. L.P. and E.V. conducted infection assays. M.J. made DNA constructs and genetically modified cell lines. P.C. and C.R. produced recombinant proteins. V.E.P. and P.C. crystallized TMEM106BLD. V.E.P. performed X-ray crystallography and prepared cryo-EM samples. N.B.C. and P.C. acquired cryo-EM data. P.C. processed cryo-EM data. V.C. and E.R. conducted HDX-MS. A.G.W. and S.R.M. conducted biolayer interferometry. T.V. made pseudoviruses, which was supervised by H.J.T. P.M. isolated and sequenced SARS-CoV-2 strains. F.D.S. provided patient-derived glioblastoma cells. A.Y. T.N. and E.B. made anti-TMEM106B antibodies. A.Y. M.R. and N.F.-H. characterized anti-TMEM106B antibodies. H.R. A.A.M. M.A.Y.-C. and E.B. conceptualized and designed anti-TMEM106B antibody production and characterization. J.B. M.J. L.P. E.V. P.C. and D.D. designed the project. J.B. P.C. and D.D. wrote initial text drafts. M.J. L.P. E.V. V.E.P. A.G.W. V.C. H.J.T. A.Y. M.R. A.A.M. M.A.Y.-C. and E.B. reviewed and edited the manuscript. P.C. and D.D. supervised the project. A.A.M. A.Y. E.B. M.R. M.A.Y.-C. N.F.-H. and T.N. are employees of Alector LLC, and H.R. was an Alector employee at the time of manuscript conception and may have an equity interest in Alector, Inc. Several authors have patents related to TMEM106B-specific antibodies. Work in the D.D. laboratory was partially funded by Alector LCC. Funding Information: We thank Niels Willems, Yanicka Smolders, Joni Punjwani, Nathalie van Winkel, Lotte Bral, Jitte Mollekens, and Kristien Minner for performing infections, qPCR, immunofluorescence, cloning, and cell line production and thank Katharina Coun, Jolien Timmermans, and Nathalie Thys for making pseudoviruses. We thank Elisabeth Heylen, Steven Carmans, and Dominique Schols for organizing central lab facilities. We thank John Skehel and David Bauer for suggestions to improve the manuscript; Kai Dallmeier and Johan Neyts for providing hamster anti- SARS-CoV-2 serum; Phil Walker and Andrew Purkiss (Francis Crick Institute) for computer, software, and synchrotron access support; and the staff of beamline I24 of the Diamond Light Source (Oxfordshire, UK) for assistance with data collection. The D.D. laboratory was supported by the Research Foundation – Flanders ( FWO ; G067423N ), Alector LCC , and patent income. The P.C. laboratory and the Structural Biology of Disease Processes Laboratory (lead by Steve Gamblin) were supported by the Francis Crick Institute , funded by Cancer Research UK ( FC001061 and FC001078 ), the UK Medical Research Council ( FC001061 and FC001078 ), and the Wellcome Trust ( FC001061 and FC001078 ). V.C. was supported by a post-doctoral position funded by King’s College London , and E.R. was supported by a UKRI Future Leaders fellowship ( MR/S015426/1 ). Cryo-EM data were collected at the London Consortium Electron Microscopy Facility at the Francis Crick Institute funded by the Wellcome Trust ( 206175/Z/17/Z to Prof. X. Zhang). Publisher Copyright: © 2023 The Author(s)

Keywords: ACE2-independent entry, SARS-CoV-2, TMEM106B, TMEM106B crystal structure, antibody neutralization, coronavirus, cryo-EM, entry receptor

Local EPrints ID: 479995

URI: http://eprints.soton.ac.uk/id/eprint/479995

DOI: doi:10.1016/j.cell.2023.06.005

ISSN: 0092-8674

PURE UUID: 32ae4b31-5253-49fb-b3ee-656c79b22e11

ORCID for Eamonn Reading: orcid.org/0000-0001-8219-0052

Date deposited: 31 Jul 2023 17:09

Last modified: 17 Mar 2024 04:19