PA3177 is a 307-residue soluble diguanylate cyclase of Pseudomonas aeruginosa with a C-terminal GGEEF domain. It is expressed mainly in biofilms, where it makes a pool of cyclic di-GMP that maintains levels and DNA binding of the transcriptional regulator BrlR and so contributes to biofilm tolerance to tobramycin and hydrogen peroxide. Loss of PA3177 lowers biofilm c-di-GMP about 5-fold but does not affect attachment, biofilm formation or planktonic motility; overexpression represses swimming and swarming in a GGEEF-dependent manner.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0005886 plasma membrane | IBA GO_REF:0000033 | MARK AS OVER ANNOTATED | Summary: IBA from PTN002895351, seeded only by E. coli transmembrane DGCs. PA3177 has no TRANSMEM feature and no hydrophobic window long enough for a TM helix; no localization data exist. Reason: The seeds' membrane location follows from TM segments that PA3177 lacks. The node-level GO_CENTRAL defect is recorded once, on wspR. Propagation Review Root cause: PROPAGATION BAD Failure modes: COMPARTMENT OR COMPLEX MISMATCH Sources checked: PANTHER:PTN002895351 · PTN002895351 SUPPORTS SOURCE BUT NOT TARGET Six E. coli inner-membrane DGC seeds (IDA, PMID:15919996); PA3177 is predicted soluble. Supporting Evidence: file:PSEAE/PA3177/PA3177-notes.md mean above 1.6, run in this session) finds no hydrophobic window |
| GO:0005886 plasma membrane | IEA GO_REF:0000044 | MARK AS OVER ANNOTATED | Summary: Mapped from the UniProt location "Cell inner membrane", which rests only on an ARBA rule. Reason: Same claim as the IBA row; no TM segment and no localization data. Supporting Evidence: file:PSEAE/PA3177/PA3177-notes.md mean above 1.6, run in this session) finds no hydrophobic window |
| GO:0043709 cell adhesion involved in single-species biofilm formation | IBA GO_REF:0000033 | REMOVE | Summary: IBA from PTN002895351 with E. coli DgcZ as sole seed (itself IMP, acts_upstream_of_or_within). For PA3177 the gene-level data contradict it: the deletion attaches and forms biofilms like wild type. Reason: Contradicted by direct evidence for this gene, and a DGC regulates rather than performs adhesion. The node-level GO_CENTRAL defect is recorded once, on wspR. Propagation Review Root cause: PROPAGATION BAD Failure modes: FUNCTIONAL DIVERGENCE ROLE CONFLATION Sources checked: UniProtKB:P31129 · E. coli DgcZ (sole seed) SUPPORTS SOURCE BUT NOT TARGET DgcZ's own annotation is IMP with qualifier acts_upstream_of_or_within (PMID:19460094). PA3177 has no attachment phenotype. Supporting Evidence: PMID:30082282 While PA3177 contributed to biofilm drug tolerance, inactivation of PA3177 had no effect on attachment and biofilm formation. |
| GO:0052621 diguanylate cyclase activity | IBA GO_REF:0000033 | ACCEPT | Summary: IBA from the PTN002895344 IBD (19 seeds, including PA3177 itself, which is expected). The IBD stands on many independent seeds. Reason: Core activity, supported by gene-specific mutant and active-site data. Supporting Evidence: PMID:30082282 Inactivation of PA3177 reduced total cellular c-di-GMP levels 5-fold under biofilm growth conditions |
| GO:0052621 diguanylate cyclase activity | IEA GO_REF:0000003 | ACCEPT | Summary: EC 2.7.7.65 mapping. Reason: Consistent with the mutant data. Supporting Evidence: PMID:30082282 Inactivation of PA3177 reduced total cellular c-di-GMP levels 5-fold under biofilm growth conditions |
| GO:0052621 diguanylate cyclase activity | IMP PMID:30082282 The PA3177 Gene Encodes an Active Diguanylate Cyclase That C... | ACCEPT | Summary: PseudoCAP seed row. Full text read: the deletion has 5-fold less biofilm c-di-GMP, complementation restores it, and the GGAAF active-site variant does not. Seed verdict SOUND. Reason: Mutant, complementation and active-site evidence for the activity. Supporting Evidence: PMID:30082282 Inactivation of PA3177 reduced total cellular c-di-GMP levels 5-fold under biofilm growth conditions PMID:30082282 multicopy expression of the variant gene PA3177_GGAAF harboring a substitution of the active-site motif, had no effect on the c-di-GMP level of PA3177 mutant biofilms |
| GO:1902201 negative regulation of bacterial-type flagellum-dependent cell motility | IBA GO_REF:0000033 | KEEP AS NON CORE | Summary: IBA from PTN002895351. For PA3177, overexpression reduces swimming and swarming in a GGEEF-dependent way, but the deletion is not hypermotile. Reason: Plausible as a capacity of PA3177-made c-di-GMP, but PA3177 is expressed mainly in biofilms and its physiological role is biofilm drug tolerance. Supporting Evidence: PMID:30082282 In contrast, overexpression of PA3177 resulted in reduced swarming, while multicopy expression of variant gene PA3177_GGAAF harboring a substitution of the active-site motif had no effect on swarming |
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Download this section (compressed HTML)Experiment: Assay purified PA3177 for c-di-GMP synthesis and localize a functional fluorescent fusion in biofilm cells, to test whether its c-di-GMP pool is spatially confined near BrlR.
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