Requirement of the RNA-binding protein SmpB during intracellular growth of Listeria monocytogenes

Mraheil, Mobarak Abu; Frantz, Renate; Teubner, Lisa; Wendt, Heiko; Linne, Uwe; Wingerath, Jessica; Wirth, Thomas; Chakraborty, Trinad

doi:10.1016/j.ijmm.2017.01.007

Cited by 6 publications

(8 citation statements)

References 42 publications

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“…However, the survival rates of the infected mice showed no significant differences between the wild type and Δ smpB strain. Previously the smpB deletion mutant of Listeria monocytogenes and Salmonella Typhimurium were avirulent and deficient in the intramacrophage survival (Kivisaar, 2003; Mraheil et al, 2017) The results were inconsistent with past reports, indicating that there are other pathways maintaining the virulence in A. veronii smpB mutants. Subsequently, sRNA AvrA was revealed as an alternative for virulence compensation.…”

Section: Discussionmentioning

confidence: 84%

“…In summary, SmpB plays an important role in protein quality control, virulence, gene transcription, and antibiotic and stress reaction (Liang and Deutscher, 2010; Li et al, 2013; Mu et al, 2013; Mraheil et al, 2017). Unlike Salmonella Typhimurium , the virulence of the A. veronii Δ smpB strain did not become avirulent but maintained a strong pathogenicity.…”

Section: Introductionmentioning

confidence: 99%

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Small RNA AvrA Regulates IscR to Increase the Stress Tolerances in SmpB Deficiency of Aeromonas veronii

Wang

et al. 2019

Front. Cell. Infect. Microbiol.

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The superbacteria Aeromonas veronii displays not only a strong pathogenicity but also the resistance to nine kinds of antibiotics, resulting in the economic losses and health hazards. Small Protein B (SmpB) plays an important role in protein quality control, virulence, and stress reactions. Transcriptomic data revealed that expressions of the type IV pilus assembly and type VI secretion system (T6SS) proteins were downregulated in SmpB deficiency, indicating that the virulence of A. veronii might be attenuated. Although SmpB deletion decreased colonization in the mouse spleen and liver, LD50 of the smpB mutant was not altered as expected, compared with the wild type. Further, the transcriptomic and quantitative RT-PCR analyses showed that the combination of the downregulated AvrA and the upregulated iron-sulfur protein activator IscR, mediated the oxidative tolerance in smpB deletion. Next a reporter plasmid was constructed in which the promoter of iscR was applied to control the expression of the enhanced green fluorescent protein (eGFP) gene. When the reporter plasmid was co-expressed with the AvrA expression into E. coli , the relative fluorescence intensity was decreased significantly, suggesting that AvrA bound to iscR mRNA by base pairing, which in turn relieved the inhibition of iscR and intensified the downstream iron-sulfur proteins. Collectively, the smpB mutant exhibited an attenuated virulence in mice and enhanced tolerances to oxidative stress. This study demonstrates the complexity of gene regulation networks mediated by sRNA in systems biology, and also reflects the strong adaptability of superbacteria A. veronii in the process of evolution.

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Section: Discussionmentioning

confidence: 84%

Section: Introductionmentioning

confidence: 99%

Small RNA AvrA Regulates IscR to Increase the Stress Tolerances in SmpB Deficiency of Aeromonas veronii

Wang

et al. 2019

Front. Cell. Infect. Microbiol.

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“…SmpB is principally considered as a ribosome rescue factor, which works with tmRNA for quality control and dissociation of stalled ribosomes. The deficient SmpB attenuates growth, colonization, virulence, and antibiotic persistence ( Mu et al, 2013 ; Liu et al, 2017 ; Mraheil et al, 2017 ). Apart from ribosome rescue, the action of SmpB alone was first shown to increase the bvgS expression as a TF without the participation of tmRNA ( Liu et al, 2015 ).…”

Section: Discussionmentioning

confidence: 99%

Peptide Aptamer PA3 Attenuates the Viability of Aeromonas veronii by Hindering of Small Protein B-Outer Membrane Protein A Signal Pathway

Liu

Chang

et al. 2022

Front. Microbiol.

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The small protein B (SmpB), previously acting as a ribosome rescue factor for translation quality control, is required for cell viability in bacteria. Here, our study reveals that SmpB possesses new function which regulates the expression of outer membrane protein A (ompA) gene as a transcription factor in Aeromonas veronii. The deletion of SmpB caused the lower transcription expression of ompA by Quantitative Real-Time PCR (qPCR). Electrophoretic mobility shift assay (EMSA) and DNase I Footprinting verified that the SmpB bound at the regions of −46 to −28 bp, −18 to +4 bp, +21 to +31 bp, and +48 to +59 bp of the predicted ompA promoter (PompA). The key sites C52AT was further identified to interact with SmpB when PompA was fused with enhanced green fluorescent protein (EGFP) and co-transformed with SmpB expression vector for the fluorescence detection, and the result was further confirmed in microscale thermophoresis (MST) assays. Besides, the amino acid sites G11S, F26I, and K152 in SmpB were the key sites for binding to PompA. In order to further develop peptide antimicrobial agents, the peptide aptamer PA3 was screened from the peptide aptamer (PA) library by bacterial two-hybrid method. The drug sensitivity test showed that PA3 effectively inhibited the growth of A. veronii. In summary, these results demonstrated that OmpA was a good drug target for A. veronii, which was regulated by the SmpB protein and the selected peptide aptamer PA3 interacted with OmpA protein to disable SmpB-OmpA signal pathway and inhibited A. veronii, suggesting that it could be used as an antimicrobial agent for the prevention and treatment of pathogens.

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“…Heat-sensitivity and different strain backgrounds could be reasons for this. Also, some reported mutants in genes that we consider as essential showed growth defects (ackA, ltaS, oppD, oppF, sipZ, sod, smpB, lmo2487) (48,50,53,54,(102)(103)(104), longer lag phases (dltA, ptsI) (105) or genetic instability (spoVG1) (106). Tn insertions in such genes could also be counter-selected because of their detrimental effects on growth.…”

Section: Discussionmentioning

confidence: 99%

Listeria monocytogenes gene essentiality under laboratory conditions and during macrophage infection

Fischer

Engelgeh

Rothe

et al. 2022

Preprint

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The Gram-positive bacterium Listeria monocytogenes occurs widespread in the environment and infects humans when ingested along with contaminated food. Such infections are particularly dangerous for risk group patients, for whom they represent a life-threatening disease. To invent novel strategies to control contamination and disease, it is important to identify those cellular processes that maintain pathogen growth in- and outside the host. We here have applied transposon insertion sequencing (Tn-Seq) to L. monocytogenes for the identification of such processes on a genome-wide scale. Our approach classified 394 open reading frames as essential for growth under standard laboratory conditions and identified 42 further genes, which become additionally essential during intracellular growth in macrophages. Most essential genes encode components of the translation machinery, act in chromosome-related processes, cell division and biosynthesis of the cellular envelope. Several cofactor biosynthesis pathways and 29 genes with unknown functions were also essential, opening novel options for the development of anti- listerial drugs. Among the genes specifically required during intracellular growth were known virulence factors, genes compensating intracellular auxotrophies and several cell division genes. Our experiments also highlight the importance of PASTA kinase signalling, glycine metabolism and chromosome segregation for efficient intracellular growth of L. monocytogenes.

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Requirement of the RNA-binding protein SmpB during intracellular growth of Listeria monocytogenes

Cited by 6 publications

References 42 publications

Small RNA AvrA Regulates IscR to Increase the Stress Tolerances in SmpB Deficiency of Aeromonas veronii

Small RNA AvrA Regulates IscR to Increase the Stress Tolerances in SmpB Deficiency of Aeromonas veronii

Peptide Aptamer PA3 Attenuates the Viability of Aeromonas veronii by Hindering of Small Protein B-Outer Membrane Protein A Signal Pathway

Listeria monocytogenes gene essentiality under laboratory conditions and during macrophage infection

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