Annotation inferences using phylogenetic trees
fleN, a gene that regulates flagellar number in Pseudomonas aeruginosa.
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fleN disruption causes multiflagellation (3-6 polar flagella), loss of motility and up-regulation of FleQ-dependent flagellar promoters; ATP binding was inferred from an N-terminal sequence motif only.
"the mutants were multiflagellate, with three to six polar flagella per bacterium as rather than one as in the wild type"
Interaction of the antiactivator FleN with the transcriptional activator FleQ regulates flagellar number in Pseudomonas aeruginosa.
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FleN interacts with FleQ in yeast two-hybrid; the Walker A K24Q mutation abolishes the interaction; ATP binding was still described as putative.
"In this report we demonstrate direct interaction of FleN and FleQ in the yeast two-hybrid system."
The FleQ protein from Pseudomonas aeruginosa functions as both a repressor and an activator to control gene expression from the pel operon promoter in response to c-di-GMP.
ATP-Induced Structural Remodeling in the Antiactivator FleN Enables Formation of the Functional Dimeric Form.
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FleN-AMP-PNP and apo crystal structures; ATP binding induces dimerization required to antagonize FleQ ATPase; hydrolysis by FleN is not required for antagonism.
"These observations posit that the dimer is the functional form of FleN and it is nucleotide binding and not hydrolysis by FleN that is necessary to exert an antagonistic effect against FleQ."
The antiactivator FleN uses an allosteric mechanism to regulate σ(54)-dependent expression of flagellar genes in Pseudomonas aeruginosa.
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FleQ AAA+ - FleN crystal structure; FleN allosterically prevents ATP binding to FleQ and abolishes FleQ ATPase activity; interface mutants are multiflagellate.
"FleN WT completely abolishes the ATPase activity of FleQFL"
Structures of the P. aeruginosa FleQ-FleN master regulators reveal large-scale conformational switching in motility and biofilm control.
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Cryo-EM FleQ-FleN complexes; FleN has weak ATPase activity, K19A reduces hydrolysis to 16% and D48A is hydrolysis-incompetent.
"was found to no longer dimerize or hydrolyze ATP efficiently (at about 16% relative to wild-type FleN)"
Diverse Partners of the Partitioning ParB Protein in Pseudomonas aeruginosa.
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FleN fluorescent fusions are evenly distributed in the cell (no polar or membrane localization).
"FleN was equally distributed in the cells independent of the terminus to which fluorescent protein was attached"
Identification of FleQ from Pseudomonas aeruginosa as a c-di-GMP-responsive transcription factor.
Regulatory Plasticity and Metabolic Trade-offs Drive Adaptive Evolution of Alternative Flagellar Configurations in Pseudomonas aeruginosa.