Genome-wide characterization of SARS-CoV-2 cytopathogenic proteins in the search of antiviral targets.
Author
Zhang, JiantaoLi, Qi
Cruz Cosme, Ruth S
Gerzanich, Vladimir
Tang, Qiyi
Simard, J Marc
Zhao, Richard Y
Date
2021-11-24Journal
bioRxiv : the Preprint Server for BiologyPublisher
Cold Spring Harbor LaboratoryType
Article
Metadata
Show full item recordAbstract
Therapeutic inhibition of critical viral functions is important for curtailing coronavirus disease-2019 (COVID-19). We sought to identify antiviral targets through genome-wide characterization of SARS-CoV-2 proteins that are crucial for viral pathogenesis and that cause harmful cytopathic effects. All twenty-nine viral proteins were tested in a fission yeast cell-based system using inducible gene expression. Twelve proteins including eight non-structural proteins (NSP1, NSP3, NSP4, NSP5, NSP6, NSP13, NSP14 and NSP15) and four accessory proteins (ORF3a, ORF6, ORF7a and ORF7b) were identified that altered cellular proliferation and integrity, and induced cell death. Cell death correlated with the activation of cellular oxidative stress. Of the twelve proteins, ORF3a was chosen for further study in mammalian cells. In human pulmonary and kidney epithelial cells, ORF3a induced cellular oxidative stress associated with apoptosis and necrosis, and caused activation of pro-inflammatory response with production of the cytokines TNF-α, IL-6, and IFN-β1, possibly through the activation of NF-κB. To further characterize the mechanism, we tested a natural ORF3a Beta variant, Q57H, and a mutant with deletion of the highly conserved residue, ΔG188. Compared to wild type ORF3a, the ΔG188 variant yielded more robust activation of cellular oxidative stress, cell death, and innate immune response. Since cellular oxidative stress and inflammation contribute to cell death and tissue damage linked to the severity of COVID-19, our findings suggest that ORF3a is a promising, novel therapeutic target against COVID-19.Keyword
ORF3aSARS-CoV-2
Schizosaccharomyces pombe
apoptosis and necrosis
cellular pro-inflammatory response
fission yeast
oxidative stress
viral therapeutic target
Identifier to cite or link to this item
http://hdl.handle.net/10713/17569ae974a485f413a2113503eed53cd6c53
10.1101/2021.11.23.469747