The bacterial toxin RelE induces specific mRNA cleavage in the A site of the eukaryote ribosome

Andreev, Dmitry E.; Hauryliuk, Vasili; Terenin, Ilya M.; Dmitriev, Sergey E.; Ehrenberg, Måns; Shatsky, Ivan N.

doi:10.1261/rna.693208

Cited by 38 publications

(32 citation statements)

References 26 publications

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“…Therefore, ectopic production of RelE rapidly shuts down translation and halts cell growth (16). Interestingly, RelE from archaea cleaves mRNA at the ribosome in E. coli, whereas RelE from E. coli cleaves mRNA positioned at mammalian and mitochondrial ribosomes (17,18).…”

mentioning

confidence: 99%

RETRACTED: Bacterial persistence by RNA endonucleases

Maisonneuve

Shakespeare

Jørgensen

et al. 2011

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

459

557

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Bacteria form persisters, individual cells that are highly tolerant to different types of antibiotics. Persister cells are genetically identical to nontolerant kin but have entered a dormant state in which they are recalcitrant to the killing activity of the antibiotics. The molecular mechanisms underlying bacterial persistence are unknown. Here, we show that the ubiquitous Lon (Long Form Filament) protease and mRNA endonucleases (mRNases) encoded by toxin-antitoxin (TA) loci are required for persistence in Escherichia coli . Successive deletion of the 10 mRNase-encoding TA loci of E . coli progressively reduced the level of persisters, showing that persistence is a phenotype common to TA loci. In all cases tested, the antitoxins, which control the activities of the mRNases, are Lon substrates. Consistently, cells lacking lon generated a highly reduced level of persisters. Moreover, Lon overproduction dramatically increased the levels of persisters in wild-type cells but not in cells lacking the 10 mRNases. These results support a simple model according to which mRNases encoded by TA loci are activated in a small fraction of growing cells by Lon-mediated degradation of the antitoxins. Activation of the mRNases, in turn, inhibits global cellular translation, and thereby induces dormancy and persistence. Many pathogenic bacteria known to enter dormant states have a plethora of TA genes. Therefore, in the future, the discoveries described here may lead to a mechanistic understanding of the persistence phenomenon in pathogenic bacteria.

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mentioning

confidence: 99%

RETRACTED: Bacterial persistence by RNA endonucleases

Maisonneuve

Shakespeare

Jørgensen

et al. 2011

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

459

557

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“…Assembly and Analysis of Translation Initiation Complexes-48 S ribosomal complexes were assembled and analyzed by toeprinting or RelE-printing assays as described earlier (14,15,18). Briefly, 48 S complexes were assembled by incubating 1 pmol of mRNA for 10 min at 30°C in a 20-l reaction volume that contained the reconstitution buffer (20 mM Tris-HCl, pH 7.5, 110 mM KOAc, 1 mM Mg(OAc) 2 , 0.25 mM spermidine-HCl, 1 mM DTT), 0.4 mM GTP-Mg(OAc) 2 and 1 mM ATP-Mg(OAc) 2 where indicated, 10 pmol of Met-tRNA i Met or deacyl-tRNA i Met or tRNA Phe , and 2.5 pmol of 40 S ribosomal subunits and combination of factors (eIF1 (10 pmol), eIF1A (10 pmol), eIF2 (5 pmol), eIF2A (5 pmol), eIF2D (5 pmol), eIF3 (5 pmol), eIF4A (10 pmol), eIF4B (5 pmol), eIF4F (2 pmol)), as described in the text.…”

Section: Methodsmentioning

confidence: 99%

“…Assembled complexes were analyzed directly by primer extension using avian myeloblastosis virus RT (Promega) essentially as described previously (11) or treated with recombinant RelE protein, followed by deproteinization, RNA precipitation, and primer extension analysis (for details, see Ref. 18). units of ApUpG ribonucleotide, and indicated amount of eIF2A or eIF2D.…”

Section: Methodsmentioning

confidence: 99%

“…1) that the partially purified fraction of RSW from RRL, which stimulated AUG-dependent Met-tRNA binding with 40 S subunits on membranes (but does not contain eIF2), also produced the toe print signal characteristic of the HCV IRES. Authenticity of the complex was supported by its sensitivity to bacterial toxin RelE, which cleaves the mRNA at the nucleotide triplet accommodated in the A-site of the ribosome only when the P-site is occupied by peptidyl-or MettRNA (18). However, unlike the signal observed with the eIF2, the appearance of the toe-print signal in that case was not affected by the presence of GTP.…”

Section: Metmentioning

confidence: 99%

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GTP-independent tRNA Delivery to the Ribosomal P-site by a Novel Eukaryotic Translation Factor

Dmitriev

Terenin

Andreev

et al. 2010

Journal of Biological Chemistry

155

177

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During translation, aminoacyl-tRNAs are delivered to the ribosome by specialized GTPases called translation factors. Here, we report the tRNA binding to the P-site of 40 S ribosomes by a novel GTP-independent factor eIF2D isolated from mammalian cells. The binding of tRNA i Met occurs after the AUG codon finds its position in the P-site of 40 S ribosomes, the situation that takes place during initiation complex formation on the hepatitis C virus internal ribosome entry site or on some other specific RNAs (leaderless mRNA and A-rich mRNAs with relaxed scanning dependence). Its activity in tRNA binding with 40 S subunits does not require the presence of the aminoacyl moiety. Moreover, the factor possesses the unique ability to deliver non-Met (elongator) tRNAs into the P-site of the 40 S subunit. The corresponding gene is found in all eukaryotes and includes an SUI1 domain present also in translation initiation factor eIF1. The versatility of translation initiation strategies in eukaryotes is discussed.

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“…Over the years, we have developed relBE of E. coli into a paradigm model system, not least because relBE homologues are abundantly present in bacterial and archaeal chromosomes [17]. Remarkably, RelE from archaea also inhibits translation by cleavage of A-site codons in E. coli [27] and RelE of E. coli cleaves mRNA at A-sites of eukaryotic 80S and mitochondrial ribosomes [36][37][38]. However, RelE homologues have not been identified in the eukaryotic domain.…”

Section: Introductionmentioning

confidence: 99%

Hypothesis: type I toxin–antitoxin genes enter the persistence field—a feedback mechanism explaining membrane homoeostasis

Gerdes

2016

Phil. Trans. R. Soc. B

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Bacteria form persisters, cells that are tolerant to multiple antibiotics and other types of environmental stress. Persister formation can be induced either stochastically in single cells of a growing bacterial ensemble, or by environmental stresses, such as nutrient starvation, in a subpopulation of cells. In many cases, the molecular mechanisms underlying persistence are still unknown. However, there is growing evidence that, in enterobacteria, both stochastically and environmentally induced persistence are controlled by the second messenger (p)ppGpp. For example, the ‘alarmone’ (p)ppGpp activates Lon, which, in turn, activates type II toxin–antitoxin (TA) modules to thereby induce persistence. Recently, it has been shown that a type I TA module, hokB / sokB , also can induce persistence. In this case, the underlying mechanism depends on the universally conserved GTPase Obg and, surprisingly, also (p)ppGpp. In the presence of (p)ppGpp, Obg stimulates hokB transcription and induces persistence. HokB toxin expression is under both negative and positive control: SokB antisense RNA inhibits hokB mRNA translation, while (p)ppGpp and Obg together stimulate hokB transcription. HokB is a small toxic membrane protein that, when produced in modest amounts, leads to membrane depolarization, cell stasis and persistence. By contrast, overexpression of HokB disrupts the membrane potential and kills the cell. These observations raise the question of how expression of HokB is regulated. Here, I propose a homoeostatic control mechanism that couples HokB expression to the membrane-bound RNase E that degrades and inactivates SokB antisense RNA. This article is part of the themed issue ‘The new bacteriology’.

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The bacterial toxin RelE induces specific mRNA cleavage in the A site of the eukaryote ribosome

Cited by 38 publications

References 26 publications

RETRACTED: Bacterial persistence by RNA endonucleases

RETRACTED: Bacterial persistence by RNA endonucleases

GTP-independent tRNA Delivery to the Ribosomal P-site by a Novel Eukaryotic Translation Factor

Hypothesis: type I toxin–antitoxin genes enter the persistence field—a feedback mechanism explaining membrane homoeostasis

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