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NEDD4 family ubiquitin ligases associate with LCMV Z’s PPXY domain and are required for virus budding, but not via direct ubiquitination of Z

Fig 5

The ubiquitin-binding, ESCRT-0 protein HGS is required during mammarenavirus infection.

(A-C) Human lung carcinoma (A549) cells were reverse transfected with small interfering (si)RNAs to the indicated targets or a non-targeting control siRNA and then two days later were infected with LCMV and cells and virus-containing cell culture media were collected after an additional two days. (A) Expression of siRNA-targeted proteins and actin was measured in cells by western blot. Infectious titers were determined by plaque assay (B) and LCMV DI particle titers were measured by plaque interfering assay (C). Data represent the mean ± SEM of three independent experiments. A one-way ANOVA with Holm-Sidak’s test for multiple comparisons was used to compare the mean values to the control siRNA in (B-D). **p< 0.01, ***p< 0.001, ****p< 0.0001.

Fig 5