Fig. 1. Ube2g2 is necessary for production of infectious virus particles.

a Schematic of the domains in Ube2g2. C89 is the catalytic cysteine mutated to lysine in this study. b Single clones of Ube2g2^−/− HeLa cells were generated using CRISPR/Cas9; a wild-type (rUbe2g2^WT) and C89K (rUbe2g2^C89K) mutant variant were reconstituted into the deletion background. Lysates were generated and resolved by gel electrophoresis to determine expression of Ube2g2 by immunoblotting. c Cellular ubiquitylation profiles were measured in lysates from rUbe2g2^WT and rUbe2g2^C89K cells by immunoblotting. d Wild-type and Ube2g2^−/− cells were infected with ZIKV and DENV at MOI 5. Extracellular virus production at indicated time points were determined by plaque assay. Error bars represent mean ± SD, n = 3 biologically independent experiments. **P < 0.01; ***P  < 0.001 (compared with wild-type by ANOVA followed by one-sided Dunnett’s test). e Wild-type and Ube2g2^−/− cells were infected with indicated coronaviruses at MOI 0.1. Extracellular virus production at 24 h post infection was determined by qPCR. Error bars represent mean ± SD, n = 4 biologically independent experiments. **P < 0.01; ***P < 0.001 (derived using unpaired Student’s t test). f Wild-type, Ube2g2^−/−, rUbe2g2^WT and rUbe2g2^C89K cells were challenged with Zika virus at MOI 5. Infectious virus production was measured using plaque assay. Error bars represent mean ± SD, n = 4 biologically independent experiments. **P < 0.01; ***P < 0.001 (compared with wild-type by ANOVA followed by one-sided Dunnett’s test). g Endogenous Ube2g2 was immunoprecipitated from mock and virus-infected cells at indicated time intervals and probed for Aup1. (Source data are provided as a Source data file).