Fig. 4: Activation of PARIS leads to cell death and inhibition of translation.
From: A virally encoded tRNA neutralizes the PARIS antiviral defence system
a, Expression of Ocr from the pBAD vector in PARIS+ culture resulted in cell toxicity, whereas the empty vector (EV) had no impact. b, Ocr expression in PARIS+ cells induced cell death (propidium iodide staining, red) and DNA compactization (DAPI staining, blue). Arrows indicate zoomed-in images of nucleoid structures from PARIS+/Ocr (top) and PARIS−/Ocr (bottom). Cells were imaged 1 h following induction of Ocr expression. c, Metabolic labelling experiments show a drop in 3H-methionine incorporation following induction of Ocr, indicative of translation inhibition following PARIS activation. d, In vitro translation in the presence of activated AriB (WT) or active site mutant (E26A). Translation of firefly luciferase mRNA was monitored by luminescence. In the presence of activated AriB, luciferase signal was reduced markedly, further indicating that PARIS functions through a mechanism of translational inhibition. c,d, Mean and s.d. were calculated for each time point for three independent experiments. Scale bars, 10 µm. a.u., Arbitrary units; 3H-t, 3H-thymidine.
