The Intrinsic Antiviral Defense to Incoming HSV-1 Genomes Includes Specific DNA Repair Proteins and Is Counteracted by the Viral Protein ICP0
Figure 3
53BP1 accumulation at sites associated with incoming HSV-1 genomes is dependent on RNF8 and RNF168.
(A) Control HepaRG cells or cells in which RNF8 had been depleted using shRNA were infected with wild-type virus at an MOI of 0.001 or ICP0-null HSV-1 at an MOI of 0.1 for 1 hr. Cells were fixed at 24 hpi, stained for ICP4, and 53BP1 localization was assessed in asymmetrically infected cells. (B) RIDDLE cells or RIDDLE cells complemented with HA-tagged RNF168 were infected and analyzed as in A. (C) HepaRG cells containing tet-inducible RNF8-GFP were induced with 0.1 µg/ml tet for 8 hrs and infected with ICP0-null HSV-1 at an MOI of 0.1. RIDDLE cells reconstituted with HA-tagged RNF168 were infected with ICP0-null HSV-1 at an MOI of 0.1. Cells were fixed at 16–24 hpi, stained for ICP4, and localization of RNF8-GFP or HA-RNF168 was assessed.