OmniVira

Loss of furin cleavage site attenuates SARS-CoV-2 pathogenesis.

Bryan A Johnson1 Xuping Xie2 Adam L Bailey3 Birte Kalveram4 Kumari G Lokugamage1 Antonio Muruato1 Jing Zou2 Xianwen Zhang2 Terry Juelich4 Jennifer K Smith4 Lihong Zhang4 Nathen Bopp4 Craig Schindewolf1 Michelle Vu1 Abigail Vanderheiden5,6 Emma S Winkler3,7 Daniele Swetnam2 Jessica A Plante1 Patricia Aguilar4 Kenneth S Plante1 Vsevolod Popov4 Benhur Lee8 Scott C Weaver1,9 Mehul S Suthar5,6,10 Andrew L Routh2 Ping Ren4 Zhiqiang Ku11 Zhiqiang An11 Kari Debbink12 Michael S Diamond3,7,13 Pei-Yong Shi2,9 Alexander N Freiberg4,9 Vineet D Menachery14,15
Affiliations 15 institutions
  1. Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA.
  2. Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX, USA.
  3. Department of Pathology and Immunology, Washington University School of Medicine, St Louis, MO, USA.
  4. Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA.
  5. Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA.
  6. Emory Vaccine Center, Emory University School of Medicine, Atlanta, GA, USA.
  7. Department of Medicine, Washington University School of Medicine, St Louis, MO, USA.
  8. Icahn School of Medicine at Mount Sinai, New York, NY, USA.
  9. Institute for Human Infection and Immunity, University of Texas Medical Branch, Galveston, TX, USA.
  10. Yerkes National Primate Research Center, Atlanta, GA, USA.
  11. Texas Therapeutics Institute, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center at Houston, Houston, USA.
  12. Department of Natural Sciences Bowie State University, Bowie, MD, USA.
  13. Department of Molecular Microbiology, Washington University School of Medicine, St Louis, MO, USA.
  14. Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA. [email protected].
  15. Institute for Human Infection and Immunity, University of Texas Medical Branch, Galveston, TX, USA. [email protected].
PMID33494095 DOI10.1038/s41586-021-03237-4 Year2021 Journal Nature Languageeng Statusppublish

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-a new coronavirus that has led to a worldwide pandemic1-has a furin cleavage site (PRRAR) in its spike protein that is absent in other group-2B coronaviruses2. To explore whether the furin cleavage site contributes to infection and pathogenesis in this virus, we generated a mutant SARS-CoV-2 that lacks the furin cleavage site (ΔPRRA). Here we report that replicates of ΔPRRA SARS-CoV-2 had faster kinetics, improved fitness in Vero E6 cells and reduced spike protein processing, as compared to parental SARS-CoV-2. However, the ΔPRRA mutant had reduced replication in a human respiratory cell line and was attenuated in both hamster and K18-hACE2 transgenic mouse models of SARS-CoV-2 pathogenesis. Despite reduced disease, the ΔPRRA mutant conferred protection against rechallenge with the parental SARS-CoV-2. Importantly, the neutralization values of sera from patients with coronavirus disease 2019 (COVID-19) and monoclonal antibodies against the receptor-binding domain of SARS-CoV-2 were lower against the ΔPRRA mutant than against parental SARS-CoV-2, probably owing to an increased ratio of particles to plaque-forming units in infections with the former. Together, our results demonstrate a critical role for the furin cleavage site in infection with SARS-CoV-2 and highlight the importance of this site for evaluating the neutralization activities of antibodies.

Supporting text Virus Host Location
Mutation 209 Amino Acid Sequence 128 Animals 1948 Antibodies, Neutralizing 80 Cell Line 158 Chlorocebus aethiops 70 COVID-19 425 Cricetinae 41 Female 289 Furin 14 Humans 1440 Lung Diseases 2 Male 224 Mice 253 Mice, Transgenic 13 Models, Molecular 99 Mutant Proteins 15 Proteolysis 5 SARS-CoV-2 453 Serine Endopeptidases 12 Spike Glycoprotein, Coronavirus 274 Vero Cells 55 Virus Replication 191 spike protein, SARS-CoV-2 157

Evidence records

1 total
Experimental Infection
1 records · 1 evidence types
Evidence type Pathogenicity Experiment
1 records
OVE4457 Pathogenicity Experiment
Key finding

ΔPRRA SARS-CoV-2 lacking the furin cleavage site was attenuated compared to parental SARS-CoV-2 in hamster and K18-hACE2 transgenic mouse models.

Virus
Severe acute respiratory syndrome coronavirus 2
Host
hamsters Cricetinae Experimental system house mouse Mus musculus Experimental system
Location
Not specified
Supporting text

The ΔPRRA mutant had reduced replication in a human respiratory cell line and was attenuated in both hamster and K18-hACE2 transgenic mouse models of SARS-CoV-2 pathogenesis.

Method
in vivo infection experiments in hamster and K18-hACE2 transgenic mouse | viral replication assays in cell culture | pathogenesis evaluation comparing mutant and parental SARS-CoV-2
Experimental system
hamster and K18-hACE2 transgenic mouse models