Projects per year
Abstract
Background.
SARS-CoV-2 is a recently emerged respiratory pathogen that has significantly impacted global human health. We wanted to rapidly characterise the transcriptomic, proteomic and phosphoproteomic landscape of this novel coronavirus to provide a fundamental description of the virus’s genomic and proteomic potential.
Methods.
We used direct RNA sequencing to determine the transcriptome of SARS-CoV-2 grown in Vero E6 cells which is widely used to propagate the novel coronavirus. The viral transcriptome was analysed using a recently developed ORF-centric pipeline. Allied to this we used tandem mass spectrometry to investigate the proteome and phosphoproteome of the same virally infected cells.
Results.
Our integrated analysis revealed that the viral transcripts (i.e. subgenomic mRNAs), generally fitted the expected transcription model for coronaviruses. Importantly, a 24 nt in-frame deletion was detected in over half of the subgenomic mRNAs encoding the spike (S) glycoprotein and was predicted to remove a proposed furin cleavage site from the S glycoprotein. Tandem mass spectrometry identified over 500 viral peptides and 44 phosphopeptides in virus infected cells, covering almost all proteins predicted to be encoded by the SARS-CoV-2 genome, including peptides unique to the deleted variant of the S glycoprotein.
Conclusions.
Detection of an apparently viable deletion in the furin cleavage site of the S glycoprotein, a leading vaccine target, shows that this and other regions of SARS-CoV-2 proteins may readily mutate. The furin site directs cleavage of the S glycoprotein into functional subunits during virus entry or exit and likely contributes strongly to the pathogenesis and zoonosis of this virus. Our data emphasises that the viral genome sequence should be carefully monitored during the growth of viral stocks for research, animal challenge models and, potentially, in clinical samples. Such variations may result in different levels of virulence, morbidity and mortality.
SARS-CoV-2 is a recently emerged respiratory pathogen that has significantly impacted global human health. We wanted to rapidly characterise the transcriptomic, proteomic and phosphoproteomic landscape of this novel coronavirus to provide a fundamental description of the virus’s genomic and proteomic potential.
Methods.
We used direct RNA sequencing to determine the transcriptome of SARS-CoV-2 grown in Vero E6 cells which is widely used to propagate the novel coronavirus. The viral transcriptome was analysed using a recently developed ORF-centric pipeline. Allied to this we used tandem mass spectrometry to investigate the proteome and phosphoproteome of the same virally infected cells.
Results.
Our integrated analysis revealed that the viral transcripts (i.e. subgenomic mRNAs), generally fitted the expected transcription model for coronaviruses. Importantly, a 24 nt in-frame deletion was detected in over half of the subgenomic mRNAs encoding the spike (S) glycoprotein and was predicted to remove a proposed furin cleavage site from the S glycoprotein. Tandem mass spectrometry identified over 500 viral peptides and 44 phosphopeptides in virus infected cells, covering almost all proteins predicted to be encoded by the SARS-CoV-2 genome, including peptides unique to the deleted variant of the S glycoprotein.
Conclusions.
Detection of an apparently viable deletion in the furin cleavage site of the S glycoprotein, a leading vaccine target, shows that this and other regions of SARS-CoV-2 proteins may readily mutate. The furin site directs cleavage of the S glycoprotein into functional subunits during virus entry or exit and likely contributes strongly to the pathogenesis and zoonosis of this virus. Our data emphasises that the viral genome sequence should be carefully monitored during the growth of viral stocks for research, animal challenge models and, potentially, in clinical samples. Such variations may result in different levels of virulence, morbidity and mortality.
Original language | English |
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Article number | 68 |
Number of pages | 15 |
Journal | Genome Medicine |
Volume | 12 |
DOIs | |
Publication status | Published - 28 Jul 2020 |
Structured keywords
- Covid19
- UNCOVER
- BrisSynBio
- Bristol BioDesign Institute
Fingerprint
Dive into the research topics of 'Characterisation of the transcriptome and proteome of SARS-CoV-2 reveals a cell passage induced in-frame deletion of the furin-like cleavage site from the spike glycoprotein'. Together they form a unique fingerprint.Projects
- 3 Finished
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Analysis of flavivirus infection on the cellular lipidome: implications for virus particle production and replication.
1/04/18 → 31/03/22
Project: Research
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UK-Indonesian Consortium to Identify Biomarkers Predictive of Dengue Disease Severity
14/11/17 → 31/03/22
Project: Research
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Adapting high throughput technologies to understand the process of viral zoonosis
4/01/16 → 3/01/20
Project: Research
Equipment
-
HPC (High Performance Computing) Facility
Sadaf R Alam (Manager), Steven A Chapman (Manager), Polly E Eccleston (Other), Simon H Atack (Other) & D A G Williams (Manager)
Facility/equipment: Facility
Profiles
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Professor Andrew D Davidson
- School of Cellular and Molecular Medicine - Professor of Systems Virology
- Infection and Immunity
Person: Academic , Member
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Dr Kate J Heesom
- Life Sciences Faculty Office - Senior Research Fellow (Proteomics Facility Director)
Person: Academic