1. Excludon lasRNA in L. monocytogenes and L. innocua.
Excludon lasRNA in L. monocytogenes and L. innocua. Long antisense RNA (yellow box‐arrows) encoded in the opposite strand of L. monocytogenes (Lmo) and L. innocua (Lin) genes (blue box‐arrows) are defined by green arrows corresponding to TSSs. The size of the transcripts (dashed blue arrow) was estimated from northern blots. (A) anti0605 is transcribed opposite to lmo0605 in L. monocytogenes and lin0614 in L. innocua. (B) Northern blots were carried out with probes specific to L. monocytogenes (shown) or L. innocua (not shown). As probes cross‐hybridized with RNA from both species, and showed identical hybridization patterns, only the blot with L. monocytogenes‐specific probe is shown. Transcription appeared to be dependent on SigB in L. monocytogenes as indicated by the failure to detect bands in the ΔsigB mutant. This SigB‐dependent regulation is likely conserved in L. innocua due to the conservation of the SigB box (σ‐B) within the promoter region (as shown). Several transcripts were detected with the longest being ∼5.8 kb and likely corresponding to the antisense RNA plus the polycistronic mRNA of lmo0606‐07‐08 operon. lmo0605 codes for MatE (NorM), an Na+‐driven multidrug efflux pump whereas the downstream operon codes for a predicted transcriptional regulator (lmo0606) and an ABC‐type multidrug transport system (lmo0607, lmo0608). (C) Fold‐change expression of the antisense transcript, lmo0605 and lmo0606‐07‐08 operon in the WT and ΔsigB mutant was measured by strand‐specific qRT–PCR analysis. In the ΔsigB mutant where the lasRNA is not expressed, expression of lmo0605 is upregulated whereas lmo0606‐07‐08 is downregulated as shown.
Omri Wurtzel et al. Mol Syst Biol 2012;8:583
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