PA0847 is an inner-membrane diguanylate cyclase with a periplasmic CHASE sensory domain between two transmembrane helices, followed by cytoplasmic HAMP, PAS and GGDEF domains. The purified PAS-GGDEF fragment synthesises cyclic di-GMP from GTP in a Mg2+-dependent reaction; the PAS domain is needed for dimerisation and high activity, and the enzyme shows little product inhibition. Deletion of PA0847 lowers cellular c-di-GMP and Psl production and increases swimming and swarming motility, and its overexpression promotes pellicle and biofilm formation. Its sensory input is unknown; mutant phenotypes respond to several amino acids and other nutrients.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0005886 plasma membrane | IEA GO_REF:0000044 | ACCEPT | Summary: Subcellular-location mapping (cell inner membrane) on an unreviewed entry. Reason: Two predicted TM helices flank a periplasmic CHASE sensor domain, with cytoplasmic HAMP, PAS and GGDEF domains; the inner membrane is the only consistent location. Supporting Evidence: file:PSEAE/PA0847/PA0847-uniprot.txt TRANSMEM 91..110 PMID:31114257 Domain analysis of PA0847 shows a unique molecular architecture with an N-terminal periplasmic sensory domain |
| GO:0006355 regulation of DNA-templated transcription | IEA GO_REF:0000002 | REMOVE | Summary: InterPro2GO from the PAS fold (IPR013767). Reason: PA0847 has no DNA-binding domain. Its PAS domain is part of the diguanylate cyclase and is needed for c-di-GMP synthesis, not transcription. The PAS-fold mapping to transcription regulation is an over-broad electronic inference for this protein. Supporting Evidence: PMID:31114257 These results indicate that the PAS domain of PA0847 is required for high levels of DGC activity |
| GO:0007165 signal transduction | IEA GO_REF:0000002 | MODIFY | Summary: InterPro2GO from the HAMP domain. True but general. Reason: The specific process is GO:0061939 c-di-GMP signaling, which the gene carries by IBA. That term is not an is_a descendant of signal transduction, so MODIFY rather than TOO_GENERAL (same treatment as tpbB). Proposed replacements: c-di-GMP signaling Supporting Evidence: PMID:32012489 Among DGC mutants, ∆PA0169, ∆PA0847, ∆PA1120 (yfiN), ∆PA3702 (wspR), and ∆PA4332 produced significantly less c‐di‐GMP compared to wild type. |
| GO:0016020 membrane | IEA GO_REF:0000002 | MODIFY | Summary: InterPro2GO membrane from the HAMP domain. Reason: Correct but general; the protein is in the inner (plasma) membrane, a term it already carries. Proposed replacements: plasma membrane Supporting Evidence: file:PSEAE/PA0847/PA0847-uniprot.txt TRANSMEM 369..392 |
| GO:0052621 diguanylate cyclase activity | IBA GO_REF:0000033 | ACCEPT | Summary: IBA from PTN005315326, which PA0847 itself seeds (expected self-seed) with tpbB, VCA0965, E. coli DgcT and Dictyostelium DgcA. Reason: Seed verdict SOUND and confirmed by a purified-protein assay; the IBD stands on the other seeds as well (see notes). Supporting Evidence: PMID:31114257 Although isolated GGDEF domain showed only residual DGC activity (data not shown), PAS-GGDEF duel domain efficiently catalyzed the formation of c-di-GMP in vitro. |
| GO:0052621 diguanylate cyclase activity | IDA PMID:16477007 Analysis of Pseudomonas aeruginosa diguanylate cyclases and ... | ACCEPT | Summary: PseudoCAP seed row for PTN005315326. PA14 cells overexpressing PA0847 held 16 pmol c-di-GMP per mg wet cell weight (Table 1; full text read on PMC), just above the 12 pmol/mg detection limit. Reason: Seed verdict SOUND. The signal is weak but positive and is a direct product measurement; the activity was later shown with purified PAS-GGDEF (PMID:31114257). Supporting Evidence: file:PSEAE/PA0847/PA0847-notes.md PA0847 GGDEF +(16) PMID:31114257 Although isolated GGDEF domain showed only residual DGC activity (data not shown), PAS-GGDEF duel domain efficiently catalyzed the formation of c-di-GMP in vitro. |
| GO:0061939 c-di-GMP signaling | IBA GO_REF:0000033 | ACCEPT | Summary: IBA from PTN005315326, seeded by Dictyostelium DgcA. Reason: PA0847 contributes to the cellular c-di-GMP pool that controls Psl production and motility. Supporting Evidence: PMID:32012489 Among DGC mutants, ∆PA0169, ∆PA0847, ∆PA1120 (yfiN), ∆PA3702 (wspR), and ∆PA4332 produced significantly less c‐di‐GMP compared to wild type. PMID:31114257 Compared with wild type strain, PA0847 mutant bacteria displayed significantly increased swimming and swarming motilities (Figure 1A and B). |
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Download this section (compressed HTML)Q: What ligand does the periplasmic CHASE domain of PA0847 sense? The mutant phenotypes shift with several amino acids, but no direct ligand is known.
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