The Spike D614G mutation increases SARS-CoV-2 infection of multiple human cell types
Abstract
A novel variant of the SARS-CoV-2 virus carrying a point mutation in the Spike protein (D614G) has recently emerged and rapidly surpassed others in prevalence. This mutation is in linkage disequilibrium with an ORF1b protein variant (P314L), making it difficult to discern the functional significance of the Spike D614G mutation from population genetics alone. Here, we perform site-directed mutagenesis on wild-type human codon optimized Spike to introduce the D614G variant. Using multiple human cell lines, including human lung epithelial cells, we found that the lentiviral particles pseudotyped with Spike D614G are more effective at transducing cells than ones pseudotyped with wild-type Spike. The increased transduction with Spike D614G ranged from 1.3 to 2.4-fold in Caco-2 and Calu-3 cells expressing endogenous ACE2, and 1.5 to 7.7-fold in A549ACE2 and Huh7.5ACE2 overexpressing ACE2. Furthermore, trans-complementation of SARS-CoV-2 virus with Spike D614G showed an increased infectivity of human cells. Although there is minimal difference in ACE2 receptor binding between the D614 and G614 Spike variants, we show that the G614 variant is more resistant to proteolytic cleavage in human cells, suggesting a possible mechanism for the increased transduction.
Data availability
All data generated or analyzed in this study are included in this published article and its supplementary information files. The Spike D614G expression plasmid has been deposited to Addgene (#166850).
Article and author information
Author details
Funding
American Heart Association (20POST35220040)
- Zharko Daniloski
Sidney Kimmel Foundation
- Neville E Sanjana
National Institute of Allergy and Infectious Diseases (R01AI123155)
- Tristan X Jordan
Pew Charitable Trusts (PEW-00033055)
- Gira Bhabha
Searle Scholars Program (SSP-2018-2737)
- Gira Bhabha
National Institute of Allergy and Infectious Diseases (R01AI147131)
- Gira Bhabha
Defense Advanced Research Projects Agency (HR0011-20-2-0040)
- Benjamin R tenOever
National Human Genome Research Institute (DP2HG010099)
- Neville E Sanjana
National Cancer Institute (R01CA218668)
- Neville E Sanjana
Defense Advanced Research Projects Agency (D18AP00053)
- Neville E Sanjana
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Frank Kirchhoff, Ulm University Medical Center, Germany
Version history
- Received: December 2, 2020
- Accepted: February 10, 2021
- Accepted Manuscript published: February 11, 2021 (version 1)
- Version of Record published: February 18, 2021 (version 2)
Copyright
© 2021, Daniloski et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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