Figure 6

The serine/threonine kinase activity sites of PRV US3 and the GTPase domain (aa 1 to 308) of GBP1 are necessary for the interaction between US3 and GBP1. A–B PRV US3 interacts with GBP1 in transfected cells. HEK293T cells were transiently transfected with the indicated plasmids for 24 h. A Cell lysates were immunoprecipitated with an anti-HA antibody, followed by immunoblotting using anti-Flag, anti-HA, or anti-GAPDH antibodies. B Representative immunofluorescent stain of DAPI (blue), Flag (green), and HA (red). Scale bar = 20 μm. C GTPase domain (aa 1 to 308) interacted with US3. In the presence of Flag-US3 plasmids, HA-GBP1, HA-GBP1(1-308), HA-GBP1(308–591), HA-GBP1(K51A), or HA-GBP1(ΔCAAX) plasmids were transfected respectively into HEK293T cells. At 24 h post-transfection, cell lysates were immunoprecipitated with an anti-HA antibody. D The kinase activity sites of US3 are required for the interaction between US3 and GBP1. HEK293T cells were transfected with the described plasmids for 24 h, then were lysed for immunoprecipitation with Flag antibodies.