Ribosome association primes the stringent factor Rel for tRNA-dependent locking in the A-site and activation of (p)ppGpp synthesis.
Acylation
Allosteric Site
Bacillus subtilis
/ genetics
Bacterial Proteins
/ metabolism
Catalytic Domain
GTP Pyrophosphokinase
/ metabolism
Guanosine Pentaphosphate
/ biosynthesis
Hydrolysis
Models, Genetic
Models, Molecular
Protein Conformation
RNA Processing, Post-Transcriptional
RNA, Transfer
/ metabolism
Ribosome Subunits, Large, Bacterial
/ metabolism
Ribosomes
/ metabolism
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
11 01 2021
11 01 2021
Historique:
accepted:
20
11
2020
revised:
18
11
2020
received:
02
11
2020
pubmed:
18
12
2020
medline:
23
1
2021
entrez:
17
12
2020
Statut:
ppublish
Résumé
In the Gram-positive Firmicute bacterium Bacillus subtilis, amino acid starvation induces synthesis of the alarmone (p)ppGpp by the RelA/SpoT Homolog factor Rel. This bifunctional enzyme is capable of both synthesizing and hydrolysing (p)ppGpp. To detect amino acid deficiency, Rel monitors the aminoacylation status of the ribosomal A-site tRNA by directly inspecting the tRNA's CCA end. Here we dissect the molecular mechanism of B. subtilis Rel. Off the ribosome, Rel predominantly assumes a 'closed' conformation with dominant (p)ppGpp hydrolysis activity. This state does not specifically select deacylated tRNA since the interaction is only moderately affected by tRNA aminoacylation. Once bound to the vacant ribosomal A-site, Rel assumes an 'open' conformation, which primes its TGS and Helical domains for specific recognition and stabilization of cognate deacylated tRNA on the ribosome. The tRNA locks Rel on the ribosome in a hyperactivated state that processively synthesises (p)ppGpp while the hydrolysis is suppressed. In stark contrast to non-specific tRNA interactions off the ribosome, tRNA-dependent Rel locking on the ribosome and activation of (p)ppGpp synthesis are highly specific and completely abrogated by tRNA aminoacylation. Binding pppGpp to a dedicated allosteric site located in the N-terminal catalytic domain region of the enzyme further enhances its synthetase activity.
Identifiants
pubmed: 33330919
pii: 6039922
doi: 10.1093/nar/gkaa1187
pmc: PMC7797070
doi:
Substances chimiques
Bacterial Proteins
0
Guanosine Pentaphosphate
38918-96-6
RNA, Transfer
9014-25-9
GTP Pyrophosphokinase
EC 2.7.6.5
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
444-457Informations de copyright
© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.
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