FIG 4.

DENV infection requires DPM1 catalytic activity. (A) Sequence alignment of the catalytic domain of Pyrococcus furiosus DPMS (pfDPMS; UniProt accession number Q8U4M3) and Homo sapiens DPM1 (hsDPM1; UniProt accession number O60762). The DAD motif is underlined in blue. (B and C) Immunoblot analysis of DPM1 expression (B) and staining of cell surface CD59 (C) in DPM1^KO 293T cells complemented with the WT or DPM1 mutated in the catalytic site (D118A and D120A). Data are representative of reults from three independent experiments. (D) Control cells, DPM1^KO 293T cells, and DPM1^KO cells complemented with the WT or catalytically dead mutants of DPM1 were inoculated with increasing MOIs of DENV2 16681. Levels of infection were quantified 48 hpi by flow cytometry using MAb 2H2. Data are means ± SD from three independent experiments performed in duplicate. Significance was calculated using a two-way ANOVA with Dunett’s multiple-comparison test. ****, P < 0.0001.