A Crystal Structure of the Dengue Virus NS5 Protein Reveals a Novel Inter-domain Interface Essential for Protein Flexibility and Virus Replication
Fig 5
Replication profiles of NS5 interface mutants.
(A) Renilla luciferase activities of DENV4 WT and mutant replicons. Equal amounts of replicon RNA (WT or mutants) were electroporated into BHK-21 cells. At the indicated time points, the transfected cells were lysed and assayed for luciferase activities. The y axis shows the log10 value of Renilla luciferase activity (RLU). Each data point is the average for three replicates, and error bars show the standard deviations. (B) 10μg in vitro transcribed infectious clone RNA was electroporated into BHK-21 cells and viral replication was monitored over a course of 5 days. Intracellular viral RNA replication as detected by qRT-PCR. The grey dotted line represents the background detection of uninfected cells. (C) Extracellular viral RNA in the supernatants detected by qRT-PCR. The grey dotted line denotes background signal of uninfected supernatant. (D) Plaque morphologies of WT and the mutants at 72 hours post electroporation. (E) IFA images showing dsRNA and NS5 co-staining and percentage infection of cells at 72 hours post electroporation.