Figure 4

HSV-1 infection is affected in cell clones selected as reovirus resistant The level of transcription and translation of the reporter gene, lacZ, present in the immediate early genes of HSV-1 is shown in A and B. A) The level of expression of mRNA is shown at 4, 8 and 16 h following infection for a library of non-reovirus selected RIE-1 cells (L42), and two clones that disrupt the Eif3s10 (p162) and S100a6 genes. The cell clone with a disrupted S100a6 gene has a dramatic increase in HSV-1 expression with a concomitant decrease in cellular gene expression by 16 h. While there is more mRNA loaded in the lane with a disrupted Eif3s10 gene than is present in the other lanes, there is no evidence for HSV-1 expression in this cell until 16 h following infection. B) Translation of the LacZ reporter in the immediate early genes of HSV-1. At 8 hours following infection, the translation of the LacZ gene is dramatically increased in clones with mutant S100a6 and Anxa2 genes, barely detectable in the population of non-selected library cells (L42) and a cell clone that tags the Aptx⁺/DnaJa1^- genes, and is not evident in the other mutant clones. C) Cell survival was determined by gentian violet staining of cells at 72 hours. L42 cells, and clones with mutations in the Aptx⁺/DnaJA1, annexin II, and S100a6 genes lysed whereas clones with mutations in the Eif3s10, Anax1, Mgat1, and Igfr2 genes were resistant to lytic infection. a-library 42, non-selected; b-Eif3s10; c-calcyclin; d-Anxa1; e-Anxa2; f-Mgat1; g-Aptx⁺/DnaJa1 (negative strand); h-Igf2r