RelA-SpoT Homolog toxins pyrophosphorylate the CCA end of tRNA to inhibit protein synthesis

Tatsuaki Kurata; Tetiana Brodiazhenko; Sofia Raquel Alves Oliveira; Mohammad Roghanian; Yuriko Sakaguchi; Kathryn Jane Turnbull; Ondřej Bulvas; Hiraku Takada; Hedvig Tamman; Andres Ainelo; Radek Pohl; Dominik Rejman; Tanel Tenson; Tsutomu Suzuki; Abel Garcia-Pino; Gemma Catherine Atkinson; Vasili Hauryliuk

doi:10.1016/j.molcel.2021.06.005

RelA-SpoT Homolog toxins pyrophosphorylate the CCA end of tRNA to inhibit protein synthesis

Tatsuaki Kurata, Tetiana Brodiazhenko, Sofia Raquel Alves Oliveira, Mohammad Roghanian, Yuriko Sakaguchi, Kathryn Jane Turnbull, Ondřej Bulvas, Hiraku Takada, Hedvig Tamman, Andres Ainelo, Radek Pohl, Dominik Rejman, Tanel Tenson, Tsutomu Suzuki, Abel Garcia-Pino, Gemma Catherine Atkinson, Vasili Hauryliuk

Department of Clinical Microbiology

31 Citations (Scopus)

Abstract

RelA-SpoT Homolog (RSH) enzymes control bacterial physiology through synthesis and degradation of the nucleotide alarmone (p)ppGpp. We recently discovered multiple families of small alarmone synthetase (SAS) RSH acting as toxins of toxin-antitoxin (TA) modules, with the FaRel subfamily of toxSAS abrogating bacterial growth by producing an analog of (p)ppGpp, (pp)pApp. Here we probe the mechanism of growth arrest used by four experimentally unexplored subfamilies of toxSAS: FaRel2, PhRel, PhRel2, and CapRel. Surprisingly, all these toxins specifically inhibit protein synthesis. To do so, they transfer a pyrophosphate moiety from ATP to the tRNA 3' CCA. The modification inhibits both tRNA aminoacylation and the sensing of cellular amino acid starvation by the ribosome-associated RSH RelA. Conversely, we show that some small alarmone hydrolase (SAH) RSH enzymes can reverse the pyrophosphorylation of tRNA to counter the growth inhibition by toxSAS. Collectively, we establish RSHs as RNA-modifying enzymes.

Original language	English
Journal	Molecular cell
Volume	81
Issue number	15
Pages (from-to)	3160-3170.e9
Number of pages	11
ISSN	1097-2765
DOIs	https://doi.org/10.1016/j.molcel.2021.06.005
Publication status	Published - 5 Aug 2021

Keywords

(p)ppApp
(p)ppGpp
RelA-SpoT Homolog
SAH
SAS
ribosome
tRNA modification
toxSAS
toxin-antitoxin
translation

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Kurata, T., Brodiazhenko, T., Alves Oliveira, S. R., Roghanian, M., Sakaguchi, Y., Turnbull, K. J., Bulvas, O., Takada, H., Tamman, H., Ainelo, A., Pohl, R., Rejman, D., Tenson, T., Suzuki, T., Garcia-Pino, A., Atkinson, G. C., & Hauryliuk, V. (2021). RelA-SpoT Homolog toxins pyrophosphorylate the CCA end of tRNA to inhibit protein synthesis. Molecular cell, 81(15), 3160-3170.e9. https://doi.org/10.1016/j.molcel.2021.06.005

Kurata, T, Brodiazhenko, T, Alves Oliveira, SR, Roghanian, M, Sakaguchi, Y, Turnbull, KJ, Bulvas, O, Takada, H, Tamman, H, Ainelo, A, Pohl, R, Rejman, D, Tenson, T, Suzuki, T, Garcia-Pino, A, Atkinson, GC & Hauryliuk, V 2021, 'RelA-SpoT Homolog toxins pyrophosphorylate the CCA end of tRNA to inhibit protein synthesis', Molecular cell, vol. 81, no. 15, pp. 3160-3170.e9. https://doi.org/10.1016/j.molcel.2021.06.005

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abstract = "RelA-SpoT Homolog (RSH) enzymes control bacterial physiology through synthesis and degradation of the nucleotide alarmone (p)ppGpp. We recently discovered multiple families of small alarmone synthetase (SAS) RSH acting as toxins of toxin-antitoxin (TA) modules, with the FaRel subfamily of toxSAS abrogating bacterial growth by producing an analog of (p)ppGpp, (pp)pApp. Here we probe the mechanism of growth arrest used by four experimentally unexplored subfamilies of toxSAS: FaRel2, PhRel, PhRel2, and CapRel. Surprisingly, all these toxins specifically inhibit protein synthesis. To do so, they transfer a pyrophosphate moiety from ATP to the tRNA 3' CCA. The modification inhibits both tRNA aminoacylation and the sensing of cellular amino acid starvation by the ribosome-associated RSH RelA. Conversely, we show that some small alarmone hydrolase (SAH) RSH enzymes can reverse the pyrophosphorylation of tRNA to counter the growth inhibition by toxSAS. Collectively, we establish RSHs as RNA-modifying enzymes.",

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T1 - RelA-SpoT Homolog toxins pyrophosphorylate the CCA end of tRNA to inhibit protein synthesis

AU - Kurata, Tatsuaki

AU - Brodiazhenko, Tetiana

AU - Alves Oliveira, Sofia Raquel

AU - Roghanian, Mohammad

AU - Sakaguchi, Yuriko

AU - Turnbull, Kathryn Jane

AU - Bulvas, Ondřej

AU - Takada, Hiraku

AU - Tamman, Hedvig

AU - Ainelo, Andres

AU - Pohl, Radek

AU - Rejman, Dominik

AU - Tenson, Tanel

AU - Suzuki, Tsutomu

AU - Garcia-Pino, Abel

AU - Atkinson, Gemma Catherine

AU - Hauryliuk, Vasili

PY - 2021/8/5

Y1 - 2021/8/5

N2 - RelA-SpoT Homolog (RSH) enzymes control bacterial physiology through synthesis and degradation of the nucleotide alarmone (p)ppGpp. We recently discovered multiple families of small alarmone synthetase (SAS) RSH acting as toxins of toxin-antitoxin (TA) modules, with the FaRel subfamily of toxSAS abrogating bacterial growth by producing an analog of (p)ppGpp, (pp)pApp. Here we probe the mechanism of growth arrest used by four experimentally unexplored subfamilies of toxSAS: FaRel2, PhRel, PhRel2, and CapRel. Surprisingly, all these toxins specifically inhibit protein synthesis. To do so, they transfer a pyrophosphate moiety from ATP to the tRNA 3' CCA. The modification inhibits both tRNA aminoacylation and the sensing of cellular amino acid starvation by the ribosome-associated RSH RelA. Conversely, we show that some small alarmone hydrolase (SAH) RSH enzymes can reverse the pyrophosphorylation of tRNA to counter the growth inhibition by toxSAS. Collectively, we establish RSHs as RNA-modifying enzymes.

AB - RelA-SpoT Homolog (RSH) enzymes control bacterial physiology through synthesis and degradation of the nucleotide alarmone (p)ppGpp. We recently discovered multiple families of small alarmone synthetase (SAS) RSH acting as toxins of toxin-antitoxin (TA) modules, with the FaRel subfamily of toxSAS abrogating bacterial growth by producing an analog of (p)ppGpp, (pp)pApp. Here we probe the mechanism of growth arrest used by four experimentally unexplored subfamilies of toxSAS: FaRel2, PhRel, PhRel2, and CapRel. Surprisingly, all these toxins specifically inhibit protein synthesis. To do so, they transfer a pyrophosphate moiety from ATP to the tRNA 3' CCA. The modification inhibits both tRNA aminoacylation and the sensing of cellular amino acid starvation by the ribosome-associated RSH RelA. Conversely, we show that some small alarmone hydrolase (SAH) RSH enzymes can reverse the pyrophosphorylation of tRNA to counter the growth inhibition by toxSAS. Collectively, we establish RSHs as RNA-modifying enzymes.

KW - (p)ppApp

KW - (p)ppGpp

KW - RelA-SpoT Homolog

KW - SAH

KW - SAS

KW - ribosome

KW - tRNA modification

KW - toxSAS

KW - toxin-antitoxin

KW - translation

UR - https://www.scopus.com/pages/publications/85111582311

U2 - 10.1016/j.molcel.2021.06.005

DO - 10.1016/j.molcel.2021.06.005

M3 - Journal article

C2 - 34174184

SN - 1097-2765

VL - 81

SP - 3160-3170.e9

JO - Molecular cell

JF - Molecular cell

IS - 15

ER -

RelA-SpoT Homolog toxins pyrophosphorylate the CCA end of tRNA to inhibit protein synthesis

Abstract

Keywords

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