Spike protein cleavage-activation in the context of the SARS-CoV-2 P681R mutation: an analysis from its first appearance in lineage A.23.1 identified in Uganda

Lubinski, B., Frazier, L. E., Phan, M. V. T., Bugembe, D. L., Cunningham, J. L., Tang, T., Daniel, S., Cotten, M. , Jaimes, J. A. and Whittaker, G. R. (2022) Spike protein cleavage-activation in the context of the SARS-CoV-2 P681R mutation: an analysis from its first appearance in lineage A.23.1 identified in Uganda. Microbiology Spectrum, 10(4), e0151422. (doi: 10.1128/spectrum.01514-22) (PMID:35766497) (PMCID:PMC9430374)

Abstract

Based on its predicted ability to affect transmissibility and pathogenesis, surveillance studies have highlighted the role of a specific mutation (P681R) in the S1/S2 furin cleavage site of the SARS-CoV-2 spike protein. Here we analyzed A.23.1, first identified in Uganda, as a P681R-containing virus several months prior to the emergence of B.1.617.2 (Delta variant). We performed assays using peptides mimicking the S1/S2 from A.23.1 and B.1.617 and observed significantly increased cleavability with furin compared to both an original B lineage (Wuhan-Hu1) and B.1.1.7 (Alpha variant). We also performed cell–cell fusion and functional infectivity assays using pseudotyped particles and observed an increase in activity for A.23.1 compared to an original B lineage spike. However, these changes in activity were not reproduced in the B lineage spike bearing only the P681R substitution. Our findings suggest that while A.23.1 has increased furin-mediated cleavage linked to the P681R substitution, this substitution needs to occur on the background of other spike protein changes to enable its functional consequences.

• Item Type: Articles
• Additional Information: This work was funded in part by the National Institute of Health research grant R01AI35270 (to G.R.W. and S.D.). We thank the global SARS-CoV-2 sequencing groups for their open and rapid sharing of sequence data and GISAID for providing an effective platform to make these data available. D.L.B., M.V.T.P., and M.C. were funded by the UK Medical Research Council (MRC/UK Research and Innovation) and the UK Department for International Development (DFID) under the MRC/DFID Concordat agreement (grant agreement no. MC_PC_20010) and Wellcome Trust, UK FCDO—Wellcome Epidemic Preparedness—Coronavirus (grant agreement no. 220977/Z/20/Z). T.T. was supported by the National Science Foundation Graduate Research Fellowship Program under grant no. DGE1650441 and the Samuel C. Fleming Family Graduate Fellowship.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Cotten, Professor Matthew

Creator Roles

Cotten, M.: Methodology, Writing – review and editing, Supervision, Funding acquisition

• Authors: Lubinski, B., Frazier, L. E., Phan, M. V. T., Bugembe, D. L., Cunningham, J. L., Tang, T., Daniel, S., Cotten, M., Jaimes, J. A., and Whittaker, G. R.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research
• Journal Name: Microbiology Spectrum
• Publisher: American Society for Microbiology
• ISSN: 2165-0497
• ISSN (Online): 2165-0497
• Published Online: 29 June 2022
• Copyright Holders: Copyright © 2022 Lubinski et al.
• First Published: First published in Microbiology Spectrum 10(4): e0151422
• Publisher Policy: Reproduced under a Creative Commons License