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. 2008 Jan 16;82(7):3342–3352. doi: 10.1128/JVI.02447-07

FIG. 2.

FIG. 2.

Mutation of Trp349 in YFV NS3 to Ala (W349A) blocks virus production. (A) Plaque phenotype analysis of YF23-W349A. Plaque assays were performed using WT or W349A YF23 transcripts. (B) Replication of YF-R.luc2A-RP-W349A. YF-R.luc2A-RP replicon transcripts, either WT or W349A, were electroporated into BHK-15 cells, and luciferase activity was measured. Data represent the results of three experiments. The error bars indicate standard deviations. (C) Packaging of YF-R.luc2A-RP-W349A by use of the trans-packaging assay. Packaging assay was performed to generate WT or W349A (bar labeled “A”) YF-R.luc2A-RP-containing PIPs. Luciferase activity is denoted as the log of RLU. The dashed line indicates mock levels of packaging. Data represent the results of three experiments. The error bars indicate standard deviations. (D) Ribbon diagram representation of the structure of the helicase domain of YFV NS3. Domain 1 is shown in green, domain 2 is brown, and domain 3 is blue. Trp349 (W349), shown in red, is present on a solvent-accessible loop in domain 2. Asp343 (D343) is also indicated in magenta. Figures were generated using Pymol (DeLano Scientific).