Interaction of Bacillus subtilis GabR with the gabTD promoter: role of repeated sequences and effect of GABA in transcriptional activation.
4-Aminobutyrate Transaminase
/ genetics
Bacillus subtilis
/ genetics
Bacterial Proteins
/ chemistry
Base Sequence
Gene Expression Regulation, Bacterial
/ drug effects
Mutation
Operon
/ genetics
Promoter Regions, Genetic
/ genetics
Protein Binding
/ drug effects
Repetitive Sequences, Nucleic Acid
/ genetics
Sequence Homology, Nucleic Acid
Succinate-Semialdehyde Dehydrogenase
/ genetics
Transcriptional Activation
/ drug effects
gamma-Aminobutyric Acid
/ metabolism
Bacillus subtilis
GabR
MocR transcription factors
pyridoxal 5′-phosphate
γ-aminobutyric acid (GABA)
Journal
The FEBS journal
ISSN: 1742-4658
Titre abrégé: FEBS J
Pays: England
ID NLM: 101229646
Informations de publication
Date de publication:
11 2020
11 2020
Historique:
received:
26
11
2019
revised:
20
02
2020
accepted:
06
03
2020
pubmed:
10
3
2020
medline:
25
5
2021
entrez:
10
3
2020
Statut:
ppublish
Résumé
Bacillus subtilis is able to use γ-aminobutyric acid (GABA) found in the soil as carbon and nitrogen source, through the action of GABA aminotransferase (GabT) and succinic semialdehyde dehydrogenase (GabD). GABA acts as molecular effector in the transcriptional activation of the gabTD operon by GabR. GabR is the most studied member of the MocR family of prokaryotic pyridoxal 5'-phosphate (PLP)-dependent transcriptional regulators, yet crucial aspects of its mechanism of action are unknown. GabR binds to the gabTD promoter, but transcription is activated only when GABA is present. Here, we demonstrated, in contrast with what had been previously proposed, that three repeated nucleotide sequences in the promoter region, two direct repeats and one inverted repeat, are specifically recognized by GabR. We carried out in vitro and in vivo experiments using mutant forms of the gabTD promoter. Our results showed that GABA activates transcription by changing the modality of interaction between GabR and the recognized sequence repeats. A hypothetical model is proposed in which GabR exists in two alternative conformations that, respectively, prevent or promote transcription. According to this model, in the absence of GABA, GabR binds to DNA interacting with all three sequence repeats, overlapping the RNA polymerase binding site and therefore preventing transcription activation. On the other hand, when GABA binds to GabR, a conformational change of the protein leads to the release of the interaction with the inverted repeat, allowing transcription initiation by RNA polymerase.
Substances chimiques
Bacterial Proteins
0
gamma-Aminobutyric Acid
56-12-2
Succinate-Semialdehyde Dehydrogenase
EC 1.2.1.24
4-Aminobutyrate Transaminase
EC 2.6.1.19
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
4952-4970Informations de copyright
© 2020 Federation of European Biochemical Societies.
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