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Negative allosteric regulation of Enterococcus faecalis small alarmone synthetase RelQ by single-stranded RNA

Beljantsev, Jelena ; Kudrin, Pavel ; Andresen, Liis ; Shingler, Victoria ; Atkinson, Gemma C. LU orcid ; Tenson, Tanel and Hauryliuk, Vasili LU orcid (2017) In Proceedings of the National Academy of Sciences of the United States of America 114(14). p.3726-3731
Abstract

The alarmone nucleotides guanosine pentaphosphate (pppGpp) and tetraphosphate (ppGpp), collectively referred to as (p)ppGpp, are key regulators of bacterial growth, stress adaptation, pathogenicity, and antibiotic tolerance. We show that the tetrameric small alarmone synthetase (SAS) RelQ from the Gram-positive pathogen Enterococcus faecalis is a sequence-specific RNA-binding protein. RelQ's enzymatic and RNA binding activities are subject to intricate allosteric regulation. (p)ppGpp synthesis is potently inhibited by the binding of single-stranded RNA. Conversely, RelQ's enzymatic activity destabilizes the RelQ:RNA complex. pppGpp, an allosteric activator of the enzyme, counteracts the effect of RNA. Tetramerization of RelQ is... (More)

The alarmone nucleotides guanosine pentaphosphate (pppGpp) and tetraphosphate (ppGpp), collectively referred to as (p)ppGpp, are key regulators of bacterial growth, stress adaptation, pathogenicity, and antibiotic tolerance. We show that the tetrameric small alarmone synthetase (SAS) RelQ from the Gram-positive pathogen Enterococcus faecalis is a sequence-specific RNA-binding protein. RelQ's enzymatic and RNA binding activities are subject to intricate allosteric regulation. (p)ppGpp synthesis is potently inhibited by the binding of single-stranded RNA. Conversely, RelQ's enzymatic activity destabilizes the RelQ:RNA complex. pppGpp, an allosteric activator of the enzyme, counteracts the effect of RNA. Tetramerization of RelQ is essential for this regulatory mechanism, because both RNA binding and enzymatic activity are abolished by deletion of the SAS-specific C-terminal helix 5α. The interplay of pppGpp binding, (p)ppGpp synthesis, and RNA binding unites two archetypal regulatory paradigms within a single protein. The mechanism is likely a prevalent but previously unappreciated regulatory switch used by the widely distributed bacterial SAS enzymes.

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author
; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
keywords
(p)ppGpp, Allosteric regulation, Nucleotide signaling, RNA-protein interaction, Stringent response
in
Proceedings of the National Academy of Sciences of the United States of America
volume
114
issue
14
pages
3726 - 3731
publisher
National Academy of Sciences
external identifiers
  • scopus:85016959996
  • pmid:28320944
ISSN
0027-8424
DOI
10.1073/pnas.1617868114
language
English
LU publication?
no
additional info
Funding Information: We thank Andreas Carlstr?m for creating the expression construct for the D82G RelQ derivative, Jose Lemos for introducing us to E. faecalis RelQ, and Elena Sineva for her valuable advice on the DRaCALA assay. This work was supported by the Estonian Research Council (Grant IUT2-22, to T.T.), the European Regional Development Fund through the Centre of Excellence for Molecular Cell Technology (V.H. and T.T.), the Estonian Science Foundation (Grant PUT37, to V.H.), the Kempe Foundation (V.H.), the Ragnar S?derberg Foundation (V.H.), and the Swedish Research Council (Grants 2013-4680, to V.H., and 2011-4791, to V.S.). Copyright: Copyright 2017 Elsevier B.V., All rights reserved.
id
55af079a-73d9-4018-bcd2-1d76e822cb43
date added to LUP
2021-09-24 20:40:53
date last changed
2025-06-16 02:26:18
@article{55af079a-73d9-4018-bcd2-1d76e822cb43,
  abstract     = {{<p>The alarmone nucleotides guanosine pentaphosphate (pppGpp) and tetraphosphate (ppGpp), collectively referred to as (p)ppGpp, are key regulators of bacterial growth, stress adaptation, pathogenicity, and antibiotic tolerance. We show that the tetrameric small alarmone synthetase (SAS) RelQ from the Gram-positive pathogen Enterococcus faecalis is a sequence-specific RNA-binding protein. RelQ's enzymatic and RNA binding activities are subject to intricate allosteric regulation. (p)ppGpp synthesis is potently inhibited by the binding of single-stranded RNA. Conversely, RelQ's enzymatic activity destabilizes the RelQ:RNA complex. pppGpp, an allosteric activator of the enzyme, counteracts the effect of RNA. Tetramerization of RelQ is essential for this regulatory mechanism, because both RNA binding and enzymatic activity are abolished by deletion of the SAS-specific C-terminal helix 5α. The interplay of pppGpp binding, (p)ppGpp synthesis, and RNA binding unites two archetypal regulatory paradigms within a single protein. The mechanism is likely a prevalent but previously unappreciated regulatory switch used by the widely distributed bacterial SAS enzymes.</p>}},
  author       = {{Beljantsev, Jelena and Kudrin, Pavel and Andresen, Liis and Shingler, Victoria and Atkinson, Gemma C. and Tenson, Tanel and Hauryliuk, Vasili}},
  issn         = {{0027-8424}},
  keywords     = {{(p)ppGpp; Allosteric regulation; Nucleotide signaling; RNA-protein interaction; Stringent response}},
  language     = {{eng}},
  number       = {{14}},
  pages        = {{3726--3731}},
  publisher    = {{National Academy of Sciences}},
  series       = {{Proceedings of the National Academy of Sciences of the United States of America}},
  title        = {{Negative allosteric regulation of Enterococcus faecalis small alarmone synthetase RelQ by single-stranded RNA}},
  url          = {{http://dx.doi.org/10.1073/pnas.1617868114}},
  doi          = {{10.1073/pnas.1617868114}},
  volume       = {{114}},
  year         = {{2017}},
}