WspR (PA3702) is a soluble response regulator diguanylate cyclase of Pseudomonas aeruginosa, the output of the Wsp surface-sensing chemosensory system. It has an N-terminal receiver domain, phosphorylated by the histidine kinase WspE, and a C-terminal GGEEF domain that condenses two GTP into cyclic di-GMP. Purified WspR makes c-di-GMP and phosphorylation stimulates it; product binding at an inhibitory site switches the enzyme between active dimers, tetramers and product-inhibited dimers. WspR-derived c-di-GMP drives the wrinkly, autoaggregative colony phenotype of wspF mutants, promotes exopolysaccharide-dependent biofilm formation and suppresses swimming and swarming motility.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0000160 phosphorelay signal transduction system | IEA GO_REF:0000002 | KEEP AS NON CORE | Summary: InterPro2GO from the response-regulator receiver domain. Reason: WspR is the receiver of the Wsp phosphorelay and phosphorylation stimulates its cyclase activity. True, but the core function is the cyclase output. Supporting Evidence: PMID:16186483 We determined that purified WspR catalyzed the formation of c-diGMP in vitro and phosphorylation stimulated this activity. |
| GO:0005886 plasma membrane | IBA GO_REF:0000033 | MARK AS OVER ANNOTATED | Summary: IBA from the PTN002895351 IBD, seeded only by six E. coli transmembrane DGCs with IDA from an inner-membrane proteome study. WspR has no TRANSMEM feature and no 19-residue hydrophobic window (scan in the notes); it is a soluble response regulator whose receiver is phosphorylated by the cytoplasmic kinase WspE. Reason: The membrane location of the E. coli seeds rests on their TM segments, which WspR lacks. The node also covers the soluble DGCs PA3177, DgcP and PA4843. A node-level GO_CENTRAL defect is recorded here for the batch. Propagation Review Root cause: PROPAGATION BAD Failure modes: COMPARTMENT OR COMPLEX MISMATCH Sources checked: PANTHER:PTN002895351 · PTN002895351 SUPPORTS SOURCE BUT NOT TARGET All six seeds (DgcN P46139, P76330, P76236, P76147, P75801, P76237) are E. coli inner-membrane DGCs with IDA from PMID:15919996 (QuickGO, 2026-11-05). Their location follows from their TM segments, not from the GGDEF domain shared with soluble members. Supporting Evidence: file:PSEAE/wspR/wspR-notes.md Kyte-Doolittle, 19-residue window, mean above 1.6: no window found in WspR (347 aa). |
| GO:0005886 plasma membrane | IEA GO_REF:0000044 | MARK AS OVER ANNOTATED | Summary: Mapped from the UniProt subcellular location "Cell inner membrane", whose only evidence in the entry is an ARBA rule. Reason: Same claim as the IBA row; WspR is a soluble response regulator. Supporting Evidence: file:PSEAE/wspR/wspR-notes.md Kyte-Doolittle, 19-residue window, mean above 1.6: no window found in WspR (347 aa). |
| GO:0042802 identical protein binding | IPI PMID:18366254 Phosphorylation-independent regulation of the diguanylate cy... | KEEP AS NON CORE | Summary: WspR self-association (dimer and tetramer) seen in crystal structures and solution studies. Reason: Oligomerization is part of how WspR activity is regulated, not its catalytic function. Supporting Evidence: PMID:18366254 By switching from an active to a product-inhibited dimer via a tetrameric assembly, WspR utilizes a novel mechanism for modulation of its activity through oligomerization. |
| GO:0043709 cell adhesion involved in single-species biofilm formation | IBA GO_REF:0000033 | MARK AS OVER ANNOTATED | Summary: IBA from PTN002895351 with a single seed, E. coli DgcZ (P31129), whose own annotation is an IMP with the qualifier acts_upstream_of_or_within (EcoCyc, PMID:19460094). The IBD asserts involved_in. Reason: A DGC makes a second messenger that regulates attachment and matrix production; it does not itself carry out cell adhesion. WspR promotes biofilm formation by raising c-di-GMP. A node-level GO_CENTRAL defect is recorded here for the batch. Propagation Review Root cause: TERM SCOPING PROBLEM Failure modes: ROLE CONFLATION Sources checked: UniProtKB:P31129 · E. coli DgcZ (sole seed) SOURCE WEAK OR INFERRED QuickGO (2026-11-05) lists GO:0043709 for DgcZ only as IMP (PMID:19460094, EcoCyc) with qualifier acts_upstream_of_or_within. Supporting Evidence: PMID:16186483 We determined that purified WspR catalyzed the formation of c-diGMP in vitro and phosphorylation stimulated this activity. |
| GO:0052621 diguanylate cyclase activity | IBA GO_REF:0000033 | ACCEPT | Summary: IBA from the PTN002895344 IBD (19 seeds: E. coli, B. subtilis, Shewanella and seven P. aeruginosa DGCs, including WspR itself, which is expected and not circular). The IBD stands on many independent seeds and the term is the defining activity of GGDEF DGCs. Reason: Core activity, shown directly for purified WspR. Supporting Evidence: PMID:16186483 We determined that purified WspR catalyzed the formation of c-diGMP in vitro and phosphorylation stimulated this activity. |
| GO:0052621 diguanylate cyclase activity | IDA PMID:16477007 Analysis of Pseudomonas aeruginosa diguanylate cyclases and ... | ACCEPT | Summary: PseudoCAP seed row. Cells overexpressing WspR accumulated 445 pmol c-di-GMP per mg wet cell weight (Table 1; full text read on PMC). Seed verdict SOUND. Reason: The paper measures c-di-GMP produced by WspR; supported further by the in vitro assay of purified WspR. Supporting Evidence: file:PSEAE/wspR/wspR-notes.md PA3702 GGEEF +(445) PMID:16186483 We determined that purified WspR catalyzed the formation of c-diGMP in vitro and phosphorylation stimulated this activity. |
| GO:0052621 diguanylate cyclase activity | IEA GO_REF:0000003 | ACCEPT | Summary: EC 2.7.7.65 mapping. Reason: Consistent with the direct assay of purified WspR. Supporting Evidence: PMID:16186483 We determined that purified WspR catalyzed the formation of c-diGMP in vitro and phosphorylation stimulated this activity. |
| GO:0052621 diguanylate cyclase activity | IMP PMID:12426335 Autolysis and autoaggregation in Pseudomonas aeruginosa colo... | ACCEPT | Summary: PseudoCAP seed row. The paper (full text read) is suppressor genetics of colony morphology and motility; it measures no c-di-GMP and mentions the cyclase function only as a hypothesis from the literature. Seed verdict INDIRECT-PHENOTYPE. Reason: The term is correct for WspR (shown in PMID:16186483), so the claim is kept, consistent with the other GO:0052621 rows. The cited paper does not show it; a PSEUDOCAP defect asks for the reference to be repointed. Supporting Evidence: PMID:16186483 We determined that purified WspR catalyzed the formation of c-diGMP in vitro and phosphorylation stimulated this activity. |
| GO:1902201 negative regulation of bacterial-type flagellum-dependent cell motility | IBA GO_REF:0000033 | ACCEPT | Summary: IBA from PTN002895351 (four E. coli DGC seeds). For WspR there is gene-level support: loss of wspR restores swimming and swarming of a wspF mutant. Reason: c-di-GMP made by active WspR represses flagellar motility. Supporting Evidence: PMID:12426335 Mutation of wspR completely suppressed colony wrinkledness and restored swarming and swimming motility |
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