Figure 4. Tse8 impacts on protein synthesis in vivo.

a, Asn tRNA synthase (asnS) can rescue Tse8 toxicity. Competition assays were performed with donors PAKΔretS or PAKΔretSΔH1 and recipient PAKΔretSΔtsei8 expressing either pJN105 or pJN:asnS. b, Cells expressing Tse8 produce less sfGfp/total cells compared to an empty vector control. This effect can be rescued by expression of Tsi8 or AsnS. sfGfp levels normalised to total cells (measured by OD₆₀₀) were monitored over time in PAKΔretSΔtsei8 with sfGFP expressed from the vacant Tn7 chromosomal site in cells containing the indicated plasmids ((+) with induction). Statistical analyses: (a) Mean CFUs/mL ± SEM of recipient cells in competition/alone are represented from represented from three independent replicates performed in triplicate (n=3). Two-tailed student’s t-test, * P<0.05; ns for PAKΔretSΔH1 [pJN105] vs PAKΔretS [pJN:asnS] (P=0.687) or vs PAKΔretSΔH1 [pJN:asnS] (P=0.631). (b) Mean fluorescent AU/OD₆₀₀ ± SEM is plotted over time from three independent replicates performed in 8 technical replicates (n=3).