Exploiting bacterial effector proteins to uncover evolutionarily conserved antiviral host machinery
Fig 3
Validation and characterization of top hits from bacterial effector screens.
A. AlphaFold predicted structures, and known or predicted enzymatic functions for indicated effector proteins identified as hits in arbovirus screens. Effector catalytic residues where substitution mutations were made are highlighted in red in AlphaFold structures. B. Representative immunoblots of Flag-tagged bacterial effector expression in LD652 cells 48 h post transfection. C-F. Fold-change in normalized viral GFP signal relative to empty vector (EV) controls 72 hpi with after transfection with indicated Flag-tagged effector constructs. Wild-type (WT) effectors are compared to their mutants (E/A or C/S). Cells were stained 72 hpi with CellTracker dye and imaged to calculate fold-change in normalized GFP signal over signals in EV treatments. Data in C-F are means ± SD; n = 3. Statistical significance was determined with unpaired student’s t-test; ns = P>0.1234, * = P<0.0332, ** = P<0.0021, *** = P<0.0002, **** = P<0.0001.