Inhibition of SRP-dependent protein secretion by the bacterial alarmone (p)ppGpp

Czech, Laura; Mais, Christopher-Nils; Kratzat, Hanna; Sarmah, Pinku; Giammarinaro, Pietro I.; Freibert, Sven‐Andreas; Esser, Hanna Folke; Musial, Joanna; Berninghausen, Otto; Steinchen, Wieland; Beckmann, Roland; Koch, Hans‐Georg; Bange, Gert

doi:10.1038/s41467-022-28675-0

Cited by 21 publications

(16 citation statements)

References 99 publications

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“…However, when YohP was incubated with U-INVs in the presence of purified SRP/FtsY, YohP was efficiently inserted into the membrane. This validates that when YohP is produced from cytosolic mRNAs, it requires the SRP pathway for targeting and insertion, which supports previous studies 53, 91 .…”

Section: Resultssupporting

confidence: 91%

“…In E. coli, there is so far no indication that SRP-and FtsY-levels are transcriptionally regulated 5 , although SRP is post-translationally regulated via proteolysis 26,106 . However, a recent study has demonstrated that under stress conditions, the hyper-phosphorylated guanine nucleotides ppGpp and pppGpp bind to the GTPase domains of both SRP and FtsY and prevent the formation of a functional targeting complex 91 . This is intriguing, because many small membrane proteins, such as YohP, are upregulated when cells encounter stress conditions [107][108][109][110] , but the simultaneous (p)ppGpp accumulation would prevent their membrane insertion by the SRP-pathway 96 .…”

Section: Discussionmentioning

confidence: 99%

“…The experiment was then repeated in the presence of norvaline, an amino acid derivative that induces leucine starvation and activates (p)ppGpp formation [93][94][95] . (p)ppGpp accumulation inhibits SRP-dependent YohP insertion by preventing SRP-FtsY complex formation 91 . In addition, (p)ppGpp accumulation allosterically inhibits RNA polymerase 96 .…”

Section: Mrna Targeting Ensures Membrane Protein Insertion During Str...mentioning

confidence: 99%

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mRNA targeting eliminates the need for the signal recognition particle during membrane protein insertion in bacteria

Sarmah

Shang

Origi

et al. 2022

Preprint

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Signal-sequence dependent protein targeting is essential for the spatiotemporal organization of eukaryotic and prokaryotic cells and facilitated by dedicated protein targeting factors, such as the signal recognition particle (SRP). However, targeting signals are not exclusively contained within proteins, but can also be present within mRNAs. By in vivo and in vitro assays, we show that mRNA targeting is controlled by the nucleotide content and by secondary structures within mRNAs. mRNA binding to bacterial membranes occurs independently of soluble targeting factors, but is dependent on the SecYEG-translocon and YidC. Importantly, membrane insertion of proteins translated from membrane-bound mRNAs occurs independently of the SRP pathway, while the latter is strictly required for proteins translated from cytosolic mRNAs. In summary, our data indicate that mRNA targeting acts in parallel to the canonical SRP-dependent protein targeting and serves as an alternative strategy for safeguarding membrane protein insertion when the SRP pathway is compromised.

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Section: Resultssupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Section: Mrna Targeting Ensures Membrane Protein Insertion During Str...mentioning

confidence: 99%

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mRNA targeting eliminates the need for the signal recognition particle during membrane protein insertion in bacteria

Sarmah

Shang

Origi

et al. 2022

Preprint

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“…However, under the influence of higher concentrations of R. palmatum root extract, S. oralis shows increasing metabolic activity, with at the same time decreasing optical density. Antibiotics or antiseptics are known to cause bacterial stress responses and metabolic adaptions [ 51 , 52 , 53 ]. The increase in metabolic activity as a stress response is a well-known finding in previous studies [ 54 , 55 ].…”

Section: Discussionmentioning

confidence: 99%

Antiplanktonic and Antibiofilm Activity of Rheum palmatum Against Streptococcus oralis and Porphyromonas gingivalis

et al. 2022

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Periodontitis and peri-implantitis are inflammatory conditions with a high global prevalence. Oral pathogens such as Porphyromonas gingivalis play a crucial role in the development of dysbiotic biofilms associated with both diseases. The aim of our study was to identify plant-derived substances which mainly inhibit the growth of “disease promoting bacteria”, by comparing the effect of Rheum palmatum root extract against P. gingivalis and the commensal species Streptococcus oralis. Antiplanktonic activity was determined by measuring optical density and metabolic activity. Antibiofilm activity was quantified using metabolic activity assays and live/dead fluorescence staining combined with confocal laser scanning microscopy. At concentrations of 3.9 mg/L, R. palmatum root extract selectively inhibited planktonic growth of the oral pathogen P. gingivalis, while not inhibiting growth of S. oralis. Selective effects also occurred in mature biofilms, as P. gingivalis was significantly more stressed and inhibited than S. oralis. Our studies show that low concentrations of R. palmatum root extract specifically inhibit P. gingivalis growth, and offer a promising approach for the development of a potential topical agent to prevent alterations in the microbiome due to overgrowth of pathogenic P. gingivalis.

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“…How does (p)ppGpp prevent chloramphenicol from killing B. subtilis? In B. subtilis, (p)ppGpp regulates many cellular processes, including DNA replication 34,35 , transcription 36 , ribosome biogenesis 37,38 and protein translation and secretion [39][40][41] . However, its most well-characterized effect in Gram-positive bacteria is the regulation of purine metabolism 31,36,37,[42][43][44][45][46][47] .…”

Section: (P)ppgpp Downregulates Gtp Levels To Prevent Chloramphenicol...mentioning

confidence: 99%

Bacillus subtilis produces (p)ppGpp in response to the bacteriostatic antibiotic chloramphenicol to prevent its potential bactericidal effect

Yang

Barra

Fung

et al. 2022

mLife

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Antibiotics combat bacteria through their bacteriostatic (by growth inhibition) or bactericidal (by killing bacteria) action. Mechanistically, it has been proposed that bactericidal antibiotics trigger cellular damage, while bacteriostatic antibiotics suppress cellular metabolism. Here, we demonstrate how the difference between bacteriostatic and bactericidal activities of the antibiotic chloramphenicol can be attributed to an antibiotic-induced bacterial protective response: the stringent response. Chloramphenicol targets the ribosome to inhibit the growth of the Gram-positive bacterium Bacillus subtilis. Intriguingly, we found that chloramphenicol becomes bactericidal in B. subtilis mutants unable to produce (p)ppGpp. We observed a similar (p)ppGpp-dependent bactericidal effect of chloramphenicol in the Gram-positive pathogen Enterococcus faecalis. In B. subtilis, chloramphenicol treatment induces (p)ppGpp accumulation through the action of the (p)ppGpp synthetase RelA. (p)ppGpp subsequently depletes the intracellular concentration of GTP and antagonizes GTP action. This GTP regulation is critical for preventing chloramphenicol from killing B. subtilis, as bypassing (p)ppGpp-dependent GTP regulation potentiates chloramphenicol killing, while reducing GTP synthesis increases survival. Finally, chloramphenicol treatment protects cells from the classical bactericidal antibiotic vancomycin, reminiscent of the clinical phenomenon of antibiotic antagonism. Taken together, our findings suggest a role of (p)ppGpp in the control of the bacteriostatic and bactericidal activity of antibiotics in Gram-positive bacteria, which can be exploited to potentiate the efficacy of existing antibiotics.

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Inhibition of SRP-dependent protein secretion by the bacterial alarmone (p)ppGpp

Cited by 21 publications

References 99 publications

mRNA targeting eliminates the need for the signal recognition particle during membrane protein insertion in bacteria

mRNA targeting eliminates the need for the signal recognition particle during membrane protein insertion in bacteria

Antiplanktonic and Antibiofilm Activity of Rheum palmatum Against Streptococcus oralis and Porphyromonas gingivalis

Bacillus subtilis produces (p)ppGpp in response to the bacteriostatic antibiotic chloramphenicol to prevent its potential bactericidal effect

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