Murine cytomegaloviruses m139 targets DDX3 to curtail interferon production and promote viral replication
Fig 5
Rescue of the MCMV m139stop replication defect in DDX3- and in UBR5-deficient SVEC4-10 cells.
(A, B) Ddx3x and Ubr5 ko SVEC4-10 cells were generated by CRISPR/Cas9 gene editing. DDX3 (A) and UBR5 (B) expression was verified by immunoblot analysis. Note that the Ddx3x ko was incomplete. (C, D, E) For multistep replication kinetics, WT (C), Ddx3x (D), and Ubr5 (E) ko SVEC4-10 cells were infected with MCMV m139-HA and MCMV m139stop (MOI = 0.01). At different days post infection, supernatants were collected for titration. Viral titers are shown as means ±SD of three biological replicates. (F, G). WT and Ddx3x ko (F) or Ubr5 ko (G) SVEC4-10 cells were infected with MCMV m139-HA or MCMV M45-HA (control) at an MOI of 5 and harvested 24 hpi. The m139 protein was immunoprecipitated using an anti-HA affinity matrix. Co-precipitating proteins were detected by immunoblotting. The immunoblots shown in this figure are representative of two (F) or three (A, B, G) independent experiments.