HipA-Triggered Growth Arrest and  -Lactam Tolerance in Escherichia coli Are Mediated by RelA-Dependent ppGpp Synthesis

Bokinsky, Gregory; Baidoo, Edward E. K.; Akella, Swetha; Burd, Helcio; Weaver, Daniel C.; Alonso-Gutierrez, Jorge; Garcia-Martin, Hector; Lee, Taek Soon; Keasling, Jay D.

doi:10.1128/jb.02210-12

Cited by 91 publications

(103 citation statements)

References 54 publications

(55 reference statements)

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“…2B) (27). Moreover, HipA also failed to induce (p)ppGpp synthesis in a ΔrelA strain (31), consistent with the fact that HipA-mediated persistence indeed depends on the activation of the stringent response in E. coli (27,28,31).…”

Section: Resultsmentioning

confidence: 56%

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RETRACTED: Stochastic induction of persister cells by HipA through (p)ppGpp-mediated activation of mRNA endonucleases

Germain

Roghanian

Gerdes

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

139

144

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The model organism Escherichia coli codes for at least 11 type II toxin-antitoxin (TA) modules, all implicated in bacterial persistence (multidrug tolerance). Ten of these encode messenger RNA endonucleases (mRNases) inhibiting translation by catalytic degradation of mRNA, and the 11th module, hipBA, encodes HipA (high persister protein A) kinase, which inhibits glutamyl tRNA synthetase (GltX). In turn, inhibition of GltX inhibits translation and induces the stringent response and persistence. Previously, we presented strong support for a model proposing (p)ppGpp (guanosine tetra and penta-phosphate) as the master regulator of persistence. Stochastic variation of [(p)ppGpp] in single cells induced TA-encoded mRNases via a pathway involving polyphosphate and Lon protease. Polyphosphate activated Lon to degrade all known type II antitoxins of E. coli. In turn, the activated mRNases induced persistence and multidrug tolerance. However, even though it was known that activation of HipA stimulated (p)ppGpp synthesis, our model did not explain how hipBA induced persistence. Here we show that, in support of and consistent with our initial model, HipA-induced persistence depends not only on (p)ppGpp but also on the 10 mRNase-encoding TA modules, Lon protease, and polyphosphate. Importantly, observations with single cells convincingly show that the high level of (p)ppGpp caused by activation of HipA does not induce persistence in the absence of TA-encoded mRNases. Thus, slow growth per se does not induce persistence in the absence of TA-encoded toxins, placing these genes as central effectors of bacterial persistence.

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Section: Resultsmentioning

confidence: 56%

“…Previously, we and others established that persistence mediated by HipA correlates with the activation of the stringent response (27,28,31). Indeed, (p)ppGpp sharply increased by over 10-fold after hipA induction ( Fig.…”

Section: Resultsmentioning

confidence: 57%

RETRACTED: Stochastic induction of persister cells by HipA through (p)ppGpp-mediated activation of mRNA endonucleases

Germain

Roghanian

Gerdes

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

139

144

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“…Using a protocol modified from ref. 61, an isocratic method was developed to isolate the ppGpp peak on an Agilent Technologies 1200 series HPLC. The buffer consisted of 0.85 M ammonium phosphate, pH 2.1.…”

Section: Methodsmentioning

confidence: 99%

The stringent response regulates adaptation to darkness in the cyanobacterium Synechococcus elongatus

Hood

Higgins

Flamholz

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

106

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The cyanobacterium Synechococcus elongatus relies upon photosynthesis to drive metabolism and growth. During darkness, Synechococcus stops growing, derives energy from its glycogen stores, and greatly decreases rates of macromolecular synthesis via unknown mechanisms. Here, we show that the stringent response, a stress response pathway whose genes are conserved across bacteria and plant plastids, contributes to this dark adaptation. Levels of the stringent response alarmone guanosine 3′-diphosphate 5′-diphosphate (ppGpp) rise after a shift from light to dark, indicating that darkness triggers the same response in cyanobacteria as starvation in heterotrophic bacteria. High levels of ppGpp are sufficient to stop growth and dramatically alter many aspects of cellular physiology, including levels of photosynthetic pigments and polyphosphate, DNA content, and the rate of translation. Cells unable to synthesize ppGpp display pronounced growth defects after exposure to darkness. The stringent response regulates expression of a number of genes in Synechococcus, including ribosomal hibernation promoting factor (hpf), which causes ribosomes to dimerize in the dark and may contribute to decreased translation. Although the metabolism of Synechococcus differentiates it from other model bacterial systems, the logic of the stringent response remains remarkably conserved, while at the same time having adapted to the unique stresses of the photosynthetic lifestyle.

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“…It was suggested that free Hip toxin increases the level of ppGpp, thereby leading to altered gene expression and thus priming cells for the phenotypic switch (131). More recently, overexpression of HipA was shown to trigger growth arrest by inducing RelA-medi-ated synthesis of ppGpp (132). Suppression of ppGpp synthesis by use of chloramphenicol relieves Hip-mediated inhibition of DNA replication, thereby restoring vulnerability to ␤-lactam antibiotics (132).…”

Section: Persisters Play a Central Role In Biofilm Recalcitrance Towamentioning

confidence: 99%

“…More recently, overexpression of HipA was shown to trigger growth arrest by inducing RelA-medi-ated synthesis of ppGpp (132). Suppression of ppGpp synthesis by use of chloramphenicol relieves Hip-mediated inhibition of DNA replication, thereby restoring vulnerability to ␤-lactam antibiotics (132).…”

Section: Persisters Play a Central Role In Biofilm Recalcitrance Towamentioning

confidence: 99%

Biofilm-Related Infections: Bridging the Gap between Clinical Management and Fundamental Aspects of Recalcitrance toward Antibiotics

Lebeaux

Ghigo

Beloin

2014

Microbiol Mol Biol Rev

1,051

862

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International audienceSurface-associated microbial communities, called biofilms, are present in all environments. Although biofilms play an important positive role in a variety of ecosystems, they also have many negative effects, including biofilm-related infections in medical settings. The ability of pathogenic biofilms to survive in the presence of high concentrations of antibiotics is called "recalcitrance" and is a characteristic property of the biofilm lifestyle, leading to treatment failure and infection recurrence. This review presents our current understanding of the molecular mechanisms of biofilm recalcitrance toward antibiotics and describes how recent progress has improved our capacity to design original and efficient strategies to prevent or eradicate biofilm-related infections

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HipA-Triggered Growth Arrest and -Lactam Tolerance in Escherichia coli Are Mediated by RelA-Dependent ppGpp Synthesis

Cited by 91 publications

References 54 publications

RETRACTED: Stochastic induction of persister cells by HipA through (p)ppGpp-mediated activation of mRNA endonucleases

RETRACTED: Stochastic induction of persister cells by HipA through (p)ppGpp-mediated activation of mRNA endonucleases

The stringent response regulates adaptation to darkness in the cyanobacterium Synechococcus elongatus

Biofilm-Related Infections: Bridging the Gap between Clinical Management and Fundamental Aspects of Recalcitrance toward Antibiotics

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