Figure 3.

Proximity-dependent labeling and pull down identify the association between mutant nsp1 and cellular proteins.A, BioID2-fused nsp1 proteins carrying mutations in the middle region of nsp1 were expressed, treated with 100 μM biotin for 16 h, and biotinylated proteins were isolated with streptavidin-coated magnetic bead. These proteins were separated in SDS gel followed by the analysis of specific protein association using immunoblot. B, the aforementioned experiment was repeated with nsp1 carrying mutations in the structurally flexible C terminus of the protein followed by immunoblot. C-terminal mutations weaken association by CPSF1, NUDT21, and eIF4G. nsp1, nonstructural protein 1.