RoeA (regulator of exopolysaccharide A, PA1107) is a 398-aa inner-membrane diguanylate cyclase of Pseudomonas aeruginosa, with five predicted transmembrane helices and a C-terminal GGDEF domain carrying the GGEEF motif. Plasmid-expressed RoeA raises cellular c-di-GMP, a GGAAF active-site mutant does not, and its isolated GGDEF domain is active in vitro when fused to PleD. In strain PA14 RoeA promotes Pel exopolysaccharide production and early biofilm formation, and a deletion swarms about twice as well as wild type without any change in flagellar function. Its effects are not explained by total c-di-GMP levels, consistent with local signalling; RoeA-GFP gives a patchy membrane pattern distinct from that of the DGC SadC. In a PAO1 deletion screen the mutant lowered global c-di-GMP but formed more biofilm biomass under flow, so the effect on biofilm depends on strain and assay.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0005886 plasma membrane | IBA GO_REF:0000033 | ACCEPT | Summary: PM IBD at PTN002895351, seeded by membrane-anchored E. coli DGCs. This protein has predicted TM helices. Reason: Consistent with the predicted topology. Supporting Evidence: file:PSEAE/PA1107/PA1107-uniprot.txt FT TRANSMEM 31..49 FT /note="Helical" FT /evidence="ECO:0000256|SAM:Phobius" |
| GO:0005886 plasma membrane | IEA GO_REF:0000044 | ACCEPT | Summary: SubCell mapping (Cell inner membrane). The UniProt statement is ARBA-derived, but the Phobius TM helices support it independently. Reason: Same claim as the IBA row. Supporting Evidence: file:PSEAE/PA1107/PA1107-uniprot.txt FT TRANSMEM 31..49 FT /note="Helical" FT /evidence="ECO:0000256|SAM:Phobius" |
| GO:0043709 cell adhesion involved in single-species biofilm formation | IBA GO_REF:0000033 | MODIFY | Summary: IBA from PTN002895351, whose GO:0043709 IBD has one seed, E. coli DgcZ, annotated acts_upstream_of_or_within. RoeA promotes Pel exopolysaccharide and biofilm formation in PA14, but it is a c-di-GMP synthase, not an adhesion component. Reason: The essence (RoeA controls biofilm formation) is sound, but GO:0043709 is a participation term and a DGC regulates the process. The replacement is GO:1900190; a signed term is not used because the deletion reduces biofilm in PA14 and increases biomass in PAO1. A node-level GO_CENTRAL defect is recorded here, once, for the batch. Propagation Review Root cause: TERM SCOPING PROBLEM Failure modes: ROLE CONFLATION Sources checked: PANTHER:PTN002895351 Β· PTN002895351 SOURCE WEAK OR INFERRED Single-seed IBD of a participation term on a DGC clade. The node-level defect is recorded once in the PA1107 review. UniProtKB:P31129 Β· E. coli DgcZ (sole seed) SUPPORTS SOURCE BUT NOT TARGET The seed annotation is IMP acts_upstream_of_or_within (EcoCyc, PMID:19460094; live QuickGO query 2026-11-05). It supports a regulatory role of DgcZ upstream of adhesion, not participation in adhesion by every DGC. For RoeA the right claim is regulation of biofilm formation. Proposed replacements: regulation of single-species biofilm formation Supporting Evidence: PMID:20978535 A deletion mutation of the PA1107 ORF was constructed (βPA1107), and a biofilm formation defect was observed at 24 h in the microtiter dish assay (Fig. 1A). PMID:20978535 Mutating the critical GGEEF motif to GGAAF resulted in loss of stimulation of Pel polysaccharide production PMID:35935233 Only four mutants exhibited elevated biomass measurements, namely PA0847 (Zhang et al., 2019), PA1107 (roeA; Cole and Lee, 2016), PA4367 and PA3825 |
| GO:0052621 diguanylate cyclase activity | IBA GO_REF:0000033 | ACCEPT | Summary: IBA from the 19-seed PTN002895344 IBD; this gene is among its own sources because of its own IDA row, which is expected. Reason: Correct; the GGEEF motif is intact and overexpression raises cellular c-di-GMP. The IBD rests on many seeds with purified-enzyme evidence. Supporting Evidence: file:PSEAE/PA1107/PA1107-notes.md PA1107 GGEEF +(35) PMID:20978535 expressing PA1107 on a plasmid significantly stimulated c-di-GMP levels versus those of the ΞPA1107 strain carrying the vector control (22-fold, P < 0.05). In contrast, expressing the PA1107-AA mutant variant did not result in a significant change in c-di-GMP levels |
| GO:0052621 diguanylate cyclase activity | IDA PMID:16477007 Analysis of Pseudomonas aeruginosa diguanylate cyclases and ... | ACCEPT | Summary: PseudoCAP IDA and a seed of PTN002895344. Extracts of the overexpressing strain contained c-di-GMP above the detection limit (Table 1, full text read on PMC). Reason: Seed verdict SOUND. A whole-cell extract of an overexpressing strain is a crude assay, but it measures the product directly and the GGEEF motif is intact. Supporting Evidence: file:PSEAE/PA1107/PA1107-notes.md PA1107 GGEEF +(35) PMID:20978535 expressing PA1107 on a plasmid significantly stimulated c-di-GMP levels versus those of the ΞPA1107 strain carrying the vector control (22-fold, P < 0.05). In contrast, expressing the PA1107-AA mutant variant did not result in a significant change in c-di-GMP levels |
| GO:0052621 diguanylate cyclase activity | IDA PMID:20978535 Specific control of Pseudomonas aeruginosa surface-associate... | ACCEPT | Summary: PseudoCAP IDA and a seed of PTN002895344. Plasmid RoeA raises c-di-GMP 22-fold in vivo and a GGAAF mutant does not; a purified PleD chimera carrying the RoeA GGDEF domain is active in vitro, and the catalytic mutant is not (full text read). Reason: Seed verdict SOUND. This is the strongest DGC evidence for RoeA: an in vitro product assay with an active-site control, plus in vivo LC-MS. Supporting Evidence: PMID:20978535 Both of these chimeras could be purified, but only the construct with the WT GGEEF domain demonstrated in vitro DGC activity (Fig. 2H). PMID:20978535 expressing PA1107 on a plasmid significantly stimulated c-di-GMP levels versus those of the ΞPA1107 strain carrying the vector control (22-fold, P < 0.05). In contrast, expressing the PA1107-AA mutant variant did not result in a significant change in c-di-GMP levels |
| GO:0052621 diguanylate cyclase activity | IEA GO_REF:0000003 | ACCEPT | Summary: EC 2.7.7.65 mapping. Reason: Same claim as the IBA and IDA rows. Supporting Evidence: file:PSEAE/PA1107/PA1107-notes.md PA1107 GGEEF +(35) PMID:20978535 expressing PA1107 on a plasmid significantly stimulated c-di-GMP levels versus those of the ΞPA1107 strain carrying the vector control (22-fold, P < 0.05). In contrast, expressing the PA1107-AA mutant variant did not result in a significant change in c-di-GMP levels |
| GO:1902201 negative regulation of bacterial-type flagellum-dependent cell motility | IBA GO_REF:0000033 | KEEP AS NON CORE | Summary: IBA from PTN002895351 (seeded by E. coli DGCs). The roeA deletion swarms about 2-fold better than wild type, and plasmid RoeA suppresses swarming, but flagellar function itself is unchanged. Reason: Supported in direction for swarming, a flagellum-dependent motility, but peripheral to RoeA's main output (Pel production). Supporting Evidence: PMID:20978535 the βroeA mutant swarms ~2-fold better than the WT PMID:20978535 Thus, it appears that mutating the roeA gene has no detectable impact on flagellar function. |
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Download this section (compressed HTML)Experiment: Test RoeA-PelD proximity (bacterial two-hybrid, co-purification) and compare roeA deletions in PA14 and PAO1 side by side in microtiter and flow-cell assays.
What is not known β curated, literature-grounded statements of the open unknowns (the inverse of core functions).
Gap: The input signal that controls RoeA, and how its c-di-GMP reaches the Pel machinery (PelD) without changing total c-di-GMP, are unknown; the sign of its effect on biofilm differs between PA14 and PAO1.
BIOLOGY
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