New SPP2002 Publications

Elevated Expression of Toxin TisB Protects Persister Cells against Ciprofloxacin but Enhances Susceptibility to Mitomycin C

Daniel EdelmannFlorian H LeinbergerNicole E SchmidMarkus OberpaulTill F SchäberleBork A Berghoff

Summary

Bacterial chromosomes harbor toxin-antitoxin (TA) systems, some of which are implicated in the formation of multidrug-tolerant persister cells. In Escherichia coli, toxin TisB from the tisB/istR-1 TA system depolarizes the inner membrane and causes ATP depletion, which presumably favors persister formation. Transcription of tisB is induced upon DNA damage due to activation of the SOS response by LexA degradation. Transcriptional activation of tisB is counteracted on the post-transcriptional level by structural features of tisB mRNA and RNA antitoxin IstR-1. Deletion of the regulatory RNA elements (mutant Δ1-41 ΔistR) uncouples TisB expression from LexA-dependent SOS induction and causes a ‘high persistence’ (hip) phenotype upon treatment with different antibiotics. Here, we demonstrate by the use of fluorescent reporters that TisB overexpression in mutant Δ1-41 ΔistR inhibits cellular processes, including the expression of SOS genes. The failure in SOS gene expression does not affect the hip phenotype upon treatment with the fluoroquinolone ciprofloxacin, likely because ATP depletion avoids strong DNA damage. By contrast, Δ1-41 ΔistR cells are highly susceptible to the DNA cross-linker mitomycin C, likely because the expression of SOS-dependent repair systems is impeded. Hence, the hip phenotype of the mutant is conditional and strongly depends on the DNA-damaging agent.

https://doi.org/10.3390/microorganisms9050943

 

Reprograming of sRNA target specificity by the leader peptide peTrpL in response to antibiotic exposure

Hendrik Melior, Siqi Li, Maximilian Stötzel, Sandra Maaß, Rubina Schütz, Saina Azarderakhsh, Aleksei Shevkoplias, Susanne Barth-Weber, Kathrin Baumgardt, John Ziebuhr, Konrad U Förstner, Zoe Chervontseva, Dörte Becher, Elena Evguenieva-Hackenberg

Summary

Trans-acting regulatory RNAs have the capacity to base pair with more mRNAs than generally detected under defined conditions, raising the possibility that sRNA target specificities vary depending on the specific metabolic or environmental conditions. In Sinorhizobium meliloti, the sRNA rnTrpL is derived from a tryptophan (Trp) transcription attenuator located upstream of the Trp biosynthesis gene trpE(G). The sRNA rnTrpL contains a small ORF, trpL, encoding the 14-aa leader peptide peTrpL. If Trp is available, efficient trpL translation causes transcription termination and liberation of rnTrpL, which subsequently acts to downregulate the trpDC operon, while peTrpL is known to have a Trp-independent role in posttranscriptional regulation of antibiotic resistance mechanisms. Here, we show that tetracycline (Tc) causes rnTrpL accumulation independently of Trp availability. In the presence of Tc, rnTrpL and peTrpL act collectively to destabilize rplUrpmA mRNA encoding ribosomal proteins L21 and L27. The three molecules, rnTrpL, peTrpL, and rplUrpmA mRNA, form an antibiotic-dependent ribonucleoprotein complex (ARNP). In vitro reconstitution of this ARNP in the presence of competing trpD and rplU transcripts revealed that peTrpL and Tc cause a shift of rnTrpL specificity towards rplU, suggesting that sRNA target prioritization may be readjusted in response to changing environmental conditions.

https://doi.org/10.1093/nar/gkab093

 

Trans-agierende Attenuator-RNA und Leaderpeptid in Bakterien

Hendrik Melior and Elena Evguenieva-Hackenberg

Summary

Bacterial transcription attenuators are a source of small RNAs (sRNAs) and leader peptides, for which no own functions were known. How-ever, the attenuator sRNA of the tryptophan (Trp) biosynthesis operon regulates gene expression in trans according to the Trp-availability. Moreover, the cognate leader peptide adopted Trp-independent functions. It builds antibiotic-dependent ribonucleoprotein complexes (ARNPs) for sRNA reprogramming and regulation of ribosomal and multiresistance genes.

doi: 10.1007/s12268­021­1545­0