Fig. 5: Ectopic expression of alpA is lethal independent of alpBCDE.

a Image shows tenfold spot dilutions of cells of the indicated strains of P. aeruginosa ectopically expressing alpA from a plasmid (pAlpA) and under the control of an IPTG inducible promoter. PAO1 indicates WT, PAO1 ΔrelA is indicated ΔrelA, PAO1 ΔrelA ΔspoT is indicated ΔrelA ΔspoT, PAO1 ΔalpBCDE is indicated ΔalpBCDE, PAO1 ΔalpBCDE ΔPA0807–PA0831 is indicated ΔalpBCDE ΔPA0807–PA0831. b, c Examples in which ectopic expression of alpA results in an increase in expression of antisense transcripts. RNA-Seq was performed with PAO1 (WT) and PAO1 ΔrelA (ΔrelA) containing pAlpA or pEV (empty vector control plasmid). Transcript abundance shown as peaks. Purple indicates transcripts aligned to the plus strand (+strand). Beige indicates transcripts aligned to the minus strand (−strand). Genes depicted as yellow arrows, arrows pointing to the right are genes encoded on the plus strand, arrows pointing to the left are genes encoded on the minus strand. Images show an increase in transcripts that align to the opposite strand the genes are encoded on (antisense) in cells containing pAlpA. This increase in antisense transcripts corresponds to transcription continuing downstream of predicted intrinsic terminator sequences as indicated with a red arrow.