FIG 6.

UL94 disrupts dimerization and translocation of MITA. (A) UL94 inhibits self-interaction of MITA. HEK293T cells (2 × 10⁶) were transfected with the indicated plasmids for 24 h. Co-immunoprecipitation and immunoblot analyses were performed with the indicated antibodies. (B) UL94 disrupts dimerization of MITA. HFFs stably expressing UL94 (2 × 10⁷) were left uninfected, infected with HCMV (MOI = 1), or transfected with HSV120 (1 μg/ml) for the indicated times. Immunoblot analyses were performed with the indicated antibodies. (C) UL94 impairs translocation of MITA to perinuclear microsome. Mita^−/− MLFs that had been reconstituted with MITA-Flag and stably transduced with UL94-HA were transfected with HSV120 (1 μg/ml) for 3 h before confocal microscopy. The aggregation ratio of MITA is shown in the histogram on the right. (D) UL94 has no effect on association of MITA with the components of translocon complex. HEK293T cells (2 × 10⁶) were transfected with the indicated plasmids for 24 h. Co-immunoprecipitation and immunoblot analyses were performed with the indicated antibodies. (E) Dimerization of MITA is required for its translocation. Mita^−/− MLFs that had been reconstituted with MITA-WT or MITA-G158L mutant were transfected with HSV120 (1 μg/ml) for 3 h before confocal microscopy.