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. 2023 Feb 8;299(3):102990. doi: 10.1016/j.jbc.2023.102990

Figure 5.

Figure 5

Mprocleaves ISG DCP1A at residue Q343.A, HEK-293T cells were transfected with N-terminal flag-tagged IFIT1, IFIT2, IFIT3, IFIT5, OSAL, OAS1, Usp18, MIX, GBP1, or DCP1A plasmids along with SARS-CoV-2 Mpro or empty vector, after 30 h of transfection, followed by Western blotting analysis. B, schematic representation of DCP1A with the position of the cleavage site. C, HEK-293T cells were separately transfected with pDCP1A-Q330A, pDCP1A-Q343A, or pDCP1A-Q351A plasmid together with SARS-CoV-2 Mpro or empty vector, protein extracts were analyzed by Western blotting with anti-flag antibody 24 h posttransfection. D, HEK-293T cells were transfected with Mpro or empty vector. Cells were lysed at 24 h after transfection and analyzed by Western blotting with anti-DCP1A antibody. DCP1A, mRNA-decapping enzyme 1a; GBP1, guanylate binding protein 1; HDAC, histone deacetylase; IFIT, interferon-induced protein with tetratricopeptide repeats; ISG, interferon-stimulated gene; MIX, mix paired-like homeobox; Mpro, main protease; OAS, 2′,5′-oligoadenylate synthetase; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; Usp18, ubiquitin specific peptidase 18.