Fig. 5

DYNLT1 is an anti-ZIKV factor that binds to ZIKV 3′UTRs. a Top, Location along DYNLT1 that binds to ZIKV. Bottom, Location along ZIKV that binds to DYNLT1. b Schematic and predicted RNA structures on ZIKV before and after DYNLT1 binding to the 3′UTR. The DYNLT1 sequence is in blue and the ZIKV sequence is in black and red. c Structure model of DYNLT1-ZIKV interaction using RNAcofold program. DYNLT1 RNA is in blue while the ZIKV genome is in red. The mutations on DYNLT1 3′UTR RNA are indicated in purple above the original bases. d EMSA data showing ZIKV and DYNLT1 RNA-RNA interactions using different amounts of WT and MT DYNLT1 3′UTR RNA. e Barplots showing the effect of DYNLT1 knockdown on ZIKV replication. Huh7 cells were transfected with 20 nM of DYNLT1 siRNA or a control siRNA. At 24 h post-transfection, cells were infected with recombinant ZIKV strain PRVABC59 at a multiplicity of infection (MOI) 0.5. The infectivities of the supernatants 1 day after infection were determined by plaque assay. Means and standard deviations (SDs) from six independent experiments are presented. f DYNLT1 knockdown reduces the replication of the ZIKV African lineage. Huh7 cells were transfected with 20 nM of DYNLT1 siRNA or a control siRNA. At 24 h post-transfection, cells were infected with a ZIKV strain Dakar containing a nanoluciferase gene. At 24 h post-infection, luciferase signals were measured. The relative luciferase signals were obtained by normalizing the luciferase readouts of the DYNLT1 siRNA-treated groups to those of the siRNA control. Means and SDs from five independent experiments are presented. g Overexpression of DYNLT1 3′UTR suppresses ZIKV replication. Huh7 cells were transfected with DYNLT 3′UTR, DYNLT1 3′UTR with mutation, or pXJ vector (100, 200, or 400 ng per well in a 24-well plate). At 24 h post-transfection, cells were infected with recombinant ZIKV strain PRVABC59 containing a nanoluciferase gene at an MOI of 0.5. At 24 h post-infection, luciferase signals were measured. The relative luciferase signals were obtained by normalizing the luciferase readouts of each group to those of the pXJ vector control. Means and SDs from four independent experiments are presented. h DYNLT1 3′UTR inhibits ZIKV replication. Huh7 cells were transfected with 200 ng of DYNLT 3′UTR, DYNLT1 3′UTR with mutation, or pXJ vector. At 24 h post-transfection, cells were infected with recombinant ZIKV strain PRVABC59 containing a nanoluciferase gene at an MOI of 0.5. At given time points, cells were harvested. The relative luciferase signals were obtained by normalizing the luciferase readouts of each group to those of control at 2 h post-infection. Means and SDs from four independent experiments are presented. i Venn diagram showing the amount of overlap between RNAs that interact with all 5 DENV and ZIKV viruses and RNAs that interact with SARS-CoV-2