Characterization of the MinD/ParA-type ATPase FlhG in Vibrio alginolyticus and implications for function of its monomeric form.
Vibrio
ATP binding
ATPase
bacterial flagellum
protein localization
Journal
Genes to cells : devoted to molecular & cellular mechanisms
ISSN: 1365-2443
Titre abrégé: Genes Cells
Pays: England
ID NLM: 9607379
Informations de publication
Date de publication:
Apr 2020
Apr 2020
Historique:
received:
27
11
2019
revised:
25
01
2020
accepted:
28
01
2020
pubmed:
6
2
2020
medline:
11
11
2020
entrez:
4
2
2020
Statut:
ppublish
Résumé
FlhG is a MinD/ParA-type ATPase that works as a negative regulator for flagellar biogenesis. In Vibrio alginolyticus, FlhG functions antagonistically with the positive regulator FlhF to generate a single polar flagellum. Here, we examined the effects of ADP and ATP on the aggregation and dimerization of Vibrio FlhG. Purified FlhG aggregated after exposure to low NaCl conditions, and its aggregation was suppressed in the presence of ADP or ATP. FlhG mutants at putative ATP-binding (K31A) or catalytic (D60A) residues showed similar aggregation profiles to the wild type, but ATP caused strong aggregation of the ATPase-stimulated D171A mutant although ADP significantly suppressed the aggregation. Results of size exclusion chromatography of purified FlhG or Vibrio cell lysates suggested that FlhG exists as a monomer in solution, and ATP does not induce FlhG dimerization. The K31A and D60A mutants eluted at monomer fractions regardless of nucleotides, but ATP shifted the elution peak of the D171A mutant to slightly earlier, presumably because of a subtle conformational change. Our results suggest that monomeric FlhG can function in vivo, whose active conformation aggregates easily.
Substances chimiques
Bacterial Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
279-287Subventions
Organisme : Japan Society for the Promotion of Science
ID : JP16H04774
Organisme : Ministry of Education, Culture, Sports, Science and Technology
ID : JP24115506
Organisme : Ministry of Education, Culture, Sports, Science and Technology
ID : JP26115705
Informations de copyright
© 2020 Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd.
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