Trafficking of Hepatitis C Virus Core Protein during Virus Particle Assembly
Figure 5
The interactions between NS2 and NS3 are essential for recruiting core from LDs.
(A) Localization of core in cells expressing ADRP-CFP electroporated with the indicated viral transcript. Panels show ADRP, core, and merged images; scale bars are 10 µm. (B) The NS2 K27A mutant shows enhanced accumulation of core around LDs. Quantification was performed as described in Materials and Methods; values show mean percent LD surface area occupied by core ± SEM (n≥10). (C) Enhanced accumulation of intracellular core in the presence of NS2 mutations (left panel) ± NS3 Q221L suppressor (right panel). Quantification was performed as described in Materials and Methods (n>38). Horizontal bars represent mean values, asterisks indicate statistically significant differences from WT (Student's t-test, p<0.05). (D–E) Dual labeling of LD-associated core and core puncta in the presence of NS2 Y39A mutation ± NS3 Q221L. Cells were electroporated with the indicated viral transcript then labeled with FlAsH followed by ReAsH after an 8 h chase. The mean number of LD-associated core (left panel) and core puncta (right panel) per cell at each timepoint were calculated as described in Materials and Methods. For all graphs, values show mean number of particles at each time point ± SEM (n = 23 to 71). For comparison with WT, please refer to Figure 4C.