Results of the Expression of ORF3a Protein of SARS-CoV2 on the levels of Activated Caspase-3 in Vero E6 cells

Analysis using flow cytometry determined an increased presence of activated caspase-3, which plays a role in caspase-dependent apoptosis, was observed in Vero E6 cells with expressed ORF3a, indicating that ORF3a of SARS-CoV2 might be able to play a role in inducing apoptosis by increasing caspase-3 levels.

Ohio State University, Columbus

The presence of specific molecules that play a role in triggering apoptosis was analyzed based on the expression of ORF3a in transfected HEK293T cells. 

Method of Determining Role of ORF3a of SARS-CoV2 in apoptosis

Ren, Y., Shu, T., Wu, D. et al. The ORF3a protein of SARS-CoV-2 induces apoptosis in cells. Cell Mol Immunol 17, 881–883 (2020). https://doi.org/10.1038/s41423-020-0485-9

The ORF3a protein of SARS-CoV-2 induces apoptosis in cells

Cleaved/activated caspase-8 was observed in the transfected cells but there was not expression of Bcl-2 indicated that ORF3a may have a pro-apoptotic function by affecting the extrinsic and not the intrinsic apoptotic pathway. 

The Role of ORF3a protein in apoptosis of host cells: Caspase-8 and Bcl-2

Cytochrome C, t-Bid, and cleaved/activated caspase-9 were observed in cells with expressed ORF3a and that if an inhibitor was used that inhibited the function of either caspase-8 or caspase-9, there was a drastic reduction in apoptosis of cells with the ORF3a protein. This further cements the proposition that ORF3a plays a role in triggering the extrinsic apoptotic pathway as well as the mechanism that links the death receptor signaling and mitochondrion-dependent apoptosis.

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