ABSTRACT
Earlier studies have shown that the d120 mutant of herpes simplex virus 1, which lacks both copies of the α4 gene, induces caspase-3-dependent apoptosis in HEp-2 cells. Apoptosis was also induced by the α4 rescuant but was blocked by the complementation of rescuant with a DNA fragment encoding the US3 protein kinase (R. Leopardi and B. Roizman, Proc. Natl. Acad. Sci. USA 93:9583–9587, 1996, and R. Leopardi, C. Van Sant, and B. Roizman, Proc. Natl. Acad. Sci. USA 94:7891–7896, 1997). To investigate its role in the apoptotic cascade, the US3 open reading frame was cloned into a baculovirus (Bac-US3) under the control of the human cytomegalovirus immediate-early promoter. We report the following. (i) Bac-US3 blocks processing of procaspase-3 to active caspase. Procaspase-3 levels remained unaltered if superinfected with Bac-US3 at 3 h afterd120 mutant infection, but significant amounts of procaspase-3 remained in cells superinfected with Bac-Us3 at 9 h postinfection with d120 mutant. (ii) The US3 protein kinase blocks the proapoptotic cascade upstream of mitochondrial involvement inasmuch as Bac-US3 blocks release of cytochrome c in cells infected with thed120 mutant. (iii) Concurrent infection of HEp-2 cells with Bac-US3 and the d120 mutant did not alter the pattern of accumulation or processing of ICP0, -22, or -27, and therefore US3 does not appear to block apoptosis by targeting these proteins.
The Virion Host Shutoff RNase Plays a Key Role in Blocking the Activation of Protein Kinase R in Cells Infected with Herpes Simplex Virus 1
