Structural basis for (p)ppGpp synthesis by the Staphylococcus aureus small alarmone synthetase RelP
Manav, Melek Cemre ; Beljantseva, Jelena ; Bojer, Martin S. ; Tenson, Tanel ; Ingmer, Hanne ; Hauryliuk, Vasili LU
and Brodersen, Ditlev E.
(2018)
In Journal of Biological Chemistry
293(9).
p.3254-3264
- Abstract
The stringent response is a global reprogramming of bacterial physiology that renders cells more tolerant to antibiotics and induces virulence gene expression in pathogens in response to stress. This process is driven by accumulation of the intracellular alarmone guanosine-5′-di(tri)phosphate-3′-diphosphate ((p)ppGpp), which is produced by enzymes of the RelA SpoT homologue (RSH) family. The Gram-positive Firmicute pathogen, Staphylococcus aureus, encodes three RSH enzymes: a multidomain RSH (Rel) that senses amino acid starvation on the ribosome and two small alarmone synthetase (SAS) enzymes, RelQ (SAS1) and RelP (SAS2). In Bacillus subtilis, RelQ (SAS1) was shown to form a tetramer that is activated by pppGpp and inhibited by... (More)
The stringent response is a global reprogramming of bacterial physiology that renders cells more tolerant to antibiotics and induces virulence gene expression in pathogens in response to stress. This process is driven by accumulation of the intracellular alarmone guanosine-5′-di(tri)phosphate-3′-diphosphate ((p)ppGpp), which is produced by enzymes of the RelA SpoT homologue (RSH) family. The Gram-positive Firmicute pathogen, Staphylococcus aureus, encodes three RSH enzymes: a multidomain RSH (Rel) that senses amino acid starvation on the ribosome and two small alarmone synthetase (SAS) enzymes, RelQ (SAS1) and RelP (SAS2). In Bacillus subtilis, RelQ (SAS1) was shown to form a tetramer that is activated by pppGpp and inhibited by single-stranded RNA, but the structural and functional regulation of RelP (SAS2) is unexplored. Here, we present crystal structures of S. aureus RelP in two major functional states, pre-catalytic (bound to GTP and the non-hydrolyzable ATP analogue, adenosine 5′-(α, β-methylene)triphosphate (AMP-CPP)), and post-catalytic (bound to pppGpp). We observed that RelP also forms a tetramer, but unlike RelQ (SAS1), it is strongly inhibited by both pppGpp and ppGpp and is insensitive to inhibition by RNA. We also identified putative metal ion-binding sites at the subunit interfaces that were consistent with the observed activation of the enzyme by Zn2+ ions. The structures reported here reveal the details of the catalytic mechanism of SAS enzymes and provide a molecular basis for understanding differential regulation of SAS enzymes in Firmicute bacteria.
(Less)
- author
- Manav, Melek Cemre
; Beljantseva, Jelena
; Bojer, Martin S.
; Tenson, Tanel
; Ingmer, Hanne
; Hauryliuk, Vasili
LU
and Brodersen, Ditlev E.
- publishing date
- 2018-03-02
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 293
- issue
- 9
- pages
- 11 pages
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- scopus:85042932610
- pmid:29326162
- ISSN
- 0021-9258
- DOI
- 10.1074/jbc.RA117.001374
- language
- English
- LU publication?
- no
- additional info
- Funding Information: This work was supported by Danish National Research Foundation Grant DNRF120 (to D. E. B.), Estonian Research Council Grant IUT2-22 (to T. T.), the European Regional Development Fund through the Centre of Excellence for Molecular Cell Technology (to V. H. and T. T.), Swedish Research Council Vetenskapsrådet Grant 2013–4680 (to V. H.), and the Ragnar Söderberg foundation (to V. H.). The authors declare that they have no conflicts of interest with the contents of this article. Publisher Copyright: © 2018 by The American Society for Biochemistry and Molecular Biology, Inc. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.
- id
- 4e3d8585-ea11-4136-b493-0d7d6a4573d7
- date added to LUP
- 2021-09-24 20:38:57
- date last changed
- 2024-06-16 19:34:44
@article{4e3d8585-ea11-4136-b493-0d7d6a4573d7,
abstract = {{<p>The stringent response is a global reprogramming of bacterial physiology that renders cells more tolerant to antibiotics and induces virulence gene expression in pathogens in response to stress. This process is driven by accumulation of the intracellular alarmone guanosine-5′-di(tri)phosphate-3′-diphosphate ((p)ppGpp), which is produced by enzymes of the RelA SpoT homologue (RSH) family. The Gram-positive Firmicute pathogen, Staphylococcus aureus, encodes three RSH enzymes: a multidomain RSH (Rel) that senses amino acid starvation on the ribosome and two small alarmone synthetase (SAS) enzymes, RelQ (SAS1) and RelP (SAS2). In Bacillus subtilis, RelQ (SAS1) was shown to form a tetramer that is activated by pppGpp and inhibited by single-stranded RNA, but the structural and functional regulation of RelP (SAS2) is unexplored. Here, we present crystal structures of S. aureus RelP in two major functional states, pre-catalytic (bound to GTP and the non-hydrolyzable ATP analogue, adenosine 5′-(α, β-methylene)triphosphate (AMP-CPP)), and post-catalytic (bound to pppGpp). We observed that RelP also forms a tetramer, but unlike RelQ (SAS1), it is strongly inhibited by both pppGpp and ppGpp and is insensitive to inhibition by RNA. We also identified putative metal ion-binding sites at the subunit interfaces that were consistent with the observed activation of the enzyme by Zn<sup>2+</sup> ions. The structures reported here reveal the details of the catalytic mechanism of SAS enzymes and provide a molecular basis for understanding differential regulation of SAS enzymes in Firmicute bacteria.</p>}},
author = {{Manav, Melek Cemre and Beljantseva, Jelena and Bojer, Martin S. and Tenson, Tanel and Ingmer, Hanne and Hauryliuk, Vasili and Brodersen, Ditlev E.}},
issn = {{0021-9258}},
language = {{eng}},
month = {{03}},
number = {{9}},
pages = {{3254--3264}},
publisher = {{American Society for Biochemistry and Molecular Biology}},
series = {{Journal of Biological Chemistry}},
title = {{Structural basis for (p)ppGpp synthesis by the Staphylococcus aureus small alarmone synthetase RelP}},
url = {{http://dx.doi.org/10.1074/jbc.RA117.001374}},
doi = {{10.1074/jbc.RA117.001374}},
volume = {{293}},
year = {{2018}},
}