fleN (PA1454, UniProt G3XD64) notes
No deep-research provider was available; the literature research below was done manually
(PubMed search "FleN Pseudomonas aeruginosa", 28 hits; key papers cached via just fetch-pmid).
Biology summary
- Identification: fleN ORF between flhF and fliA encodes a 280-aa protein "with an ATP-binding
motif at its N terminus"; insertion mutants are multiflagellate and FleQ-dependent flagellar
promoters are up-regulated PMID:10629180. Full text of this paper is not
available to us (abstract only; PMC publisher-restricted).
- FleN-FleQ interaction (yeast two-hybrid), Walker A K24Q abolishes it; ATP binding still described as
putative PMID:11673434.
- FleN with FleQ at the pel promoter: FleN binds DNA-bound FleQ and bends DNA; requires ATP but not
hydrolysis PMID:22581773.
- Crystal structures (apo and AMP-PNP): ATP binding remodels FleN into a dimer; nucleotide binding,
not hydrolysis, is needed to antagonize FleQ PMID:28065505.
- FleQ AAA+ / FleN complex: FleN allosterically blocks ATP binding to FleQ PMID:34669473; FleN has its own weak ATPase activity
PMID:34669473.
- Cryo-EM FleQ-FleN (+/- c-di-GMP): K19A "no longer dimerize or hydrolyze ATP efficiently (at about 16%
relative to wild-type FleN)"; D48A is "ATP-binding but hydrolysis-incompetent" PMID:38051770.
- Localization: FleN fluorescent fusions diffuse PMID:36622167. No evidence of the
MinD-like membrane association.
Decisions
- ATP binding IDA (PseudoCAP): term true, evidence code likely wrong (sequence motif). ACCEPT + PSEUDOCAP
source defect (WRONG_EVIDENCE_CODE, PROBABLE, CHECK).
- ATP binding IBA: ACCEPT; GO_CENTRAL source defect asking to re-seed PTN000344128.
- ATP hydrolysis activity IBA: ACCEPT (weak intrinsic ATPase shown, PMID:38051770, PMID:34669473).
- cytosol IBA: ACCEPT. cytoplasmic side of plasma membrane IBA: MARK_AS_OVER_ANNOTATED (MinD-specific).
- NEW: GO:0042030 ATPase inhibitor activity (IDA, PMID:34669473) for anti-activation of FleQ;
GO:1902209 negative regulation of bacterial-type flagellum assembly (IMP, PMID:10629180). FleN performs
the regulatory step itself (binds/inhibits FleQ), so the participation test is met.
PseudoCAP IBA-seed assessment
| PTN node |
node term |
n_seeds |
leaf IBAs |
seed annotation (term, evidence, PMID) |
verdict |
rationale |
| PTN000344128 |
GO:0005524 ATP binding |
1 (sole seed) |
149 (75 taxa) |
GO:0005524 ATP binding, IDA, PMID:10629180 |
WRONG (evidence code; term itself correct) |
PMID:10629180 only reports an ATP-binding sequence motif; the same authors called FleN "putative ATP/GTP binding" in 2001 (PMID:11673434). Not direct evidence, so the row should be ISM/ISS or re-keyed to PMID:28065505 (FleN-AMP-PNP structure). The IBD conclusion (MinD/ParA/FlhG clade binds ATP) is biologically sound and the leaf IBAs are not wrong, but the node needs a genuine experimental seed (FleN PMID:28065505 or E. coli MinD) before the PseudoCAP row is corrected. |
Caveat: full text of PMID:10629180 was not accessible; the evidence-code verdict rests on the abstract
plus the authors' 2001 wording, hence confidence PROBABLE.