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Literature detail

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].
PMID 33494095 2021 Nature eng ppublish
PubMed DOI Browse context

Article

Publication summary

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-a new coronavirus that has led to a worldwide pandemic<sup>1</sup>-has a furin cleavage site (PRRAR) in its spike protein that is absent in other group-2B coronaviruses<sup>2</sup>. 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.

Mutation Amino Acid Sequence Animals Antibodies, Neutralizing Cell Line Chlorocebus aethiops COVID-19 Cricetinae Female Furin Humans Lung Diseases Male Mice Mice, Transgenic Models, Molecular Mutant Proteins Proteolysis

Structured evidence records

Evidence records

6 total
Host Range Experiment 4 records
Host Range Experiment Extraction confidence 0.95
Key finding

ΔPRRA mutant SARS-CoV-2 replicated more efficiently than parental virus in Vero E6 cells.

Virus
SARS-CoV-2
Location
Not specified
Supporting text

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.

Method
replication assay
Experimental system
in vitro cell culture
Host Range Experiment Extraction confidence 0.95
Key finding

ΔPRRA mutant SARS-CoV-2 showed reduced replication in a human respiratory cell line compared to parental virus.

Virus
SARS-CoV-2
Location
Not specified
Supporting text

The ΔPRRA mutant had reduced replication in a human respiratory cell line.

Method
replication assay
Experimental system
in vitro cell culture
Host Range Experiment Extraction confidence 0.95
Key finding

In vivo experiments showed that the ΔPRRA SARS-CoV-2 mutant was attenuated in hamsters and K18-hACE2 transgenic mice.

Virus
SARS-CoV-2
Host
Cricetinae
Location
Not specified
Supporting text

The ΔPRRA mutant was attenuated in both hamster and K18-hACE2 transgenic mouse models of SARS-CoV-2 pathogenesis.

Method
experimental infection
Experimental system
in vivo animal experiment
Host Range Experiment Extraction confidence 0.95
Key finding

In vivo experiments demonstrated attenuation of ΔPRRA SARS-CoV-2 infection in K18-hACE2 transgenic mice compared to the parental strain.

Virus
SARS-CoV-2
Location
Not specified
Supporting text

The ΔPRRA mutant was attenuated in both hamster and K18-hACE2 transgenic mouse models of SARS-CoV-2 pathogenesis.

Method
experimental infection
Experimental system
in vivo animal experiment
Molecular Adaptation 1 records
Molecular Adaptation Extraction confidence 0.95
Key finding

Deletion of the furin cleavage site (PRRAR) in the SARS-CoV-2 spike protein reduces replication efficiency in human cells and attenuates pathogenesis, indicating this site is a molecular adaptation contributing to viral infectivity and virulence.

Virus
SARS-CoV-2
Host
Not specified
Location
Not specified
Supporting text

SARS-CoV-2 has a furin cleavage site (PRRAR) in its spike protein that is absent in other group-2B coronaviruses. A mutant SARS-CoV-2 lacking the furin cleavage site (ΔPRRA) showed faster kinetics in Vero E6 cells but reduced replication in a human respiratory cell line and was attenuated in hamster and K18-hACE2 mouse models, demonstrating a critical role for the furin cleavage site in infection and pathogenesis.

Genes or proteins
spike protein
Mutations
ΔPRRA
Mechanism types
pathogenicity; replication_efficiency
Receptor Usage 1 records
Receptor Usage Extraction confidence 0.80
Key finding

Deletion of the spike protein furin cleavage site in SARS-CoV-2 reduces spike processing and viral entry efficiency in human respiratory cells, demonstrating the cleavage site's importance in receptor-mediated infection.

Virus
SARS-CoV-2
Location
Not specified
Supporting text

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has a furin cleavage site (PRRAR) in its spike protein; a ΔPRRA SARS-CoV-2 mutant lacking this site had reduced spike protein processing and reduced replication in a human respiratory cell line, indicating the furin cleavage site's role in infection.

Receptors
furin cleavage site