PA2133 is a small (285-aa) stand-alone EAL-domain protein with no predicted transmembrane segment; its catalytic motif reads ETL. It is an active c-di-GMP-specific phosphodiesterase. Lysates of cells overexpressing PA2133 degrade added c-di-GMP, and expressing PA2133 in P. aeruginosa lowers cellular c-di-GMP below detection, even in a wspF mutant with a hyperactive WspR diguanylate cyclase. Because it is so effective, PA2133 is widely used as an experimental c-di-GMP sink: its expression blocks attachment and biofilm formation, disperses established biofilms and lowers type III secretion-dependent cytotoxicity. A PA2133 loss-of-function mutant has no attachment phenotype, and the physiological role and regulation of the native protein are unknown.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0005886 plasma membrane | IBA GO_REF:0000033 | MARK AS OVER ANNOTATED | Summary: PM IBD at PTN002105365, seeded by membrane-anchored E. coli EAL proteins. PA2133 is a 285-aa lone EAL domain with no predicted TM segment. Reason: No evidence that PA2133 is membrane-associated, and its architecture suggests a soluble protein. Peripheral membrane association is not excluded. The node-level defect (PM term on a clade with soluble members) is recorded once in the rocR review. Propagation Review Root cause: PROPAGATION BAD Failure modes: COMPARTMENT OR COMPLEX MISMATCH Sources checked: PANTHER:PTN002105365 · PTN002105365 SUPPORTS SOURCE BUT NOT TARGET Membrane seeds are TM proteins; PA2133 has no TM segment. |
| GO:0071111 cyclic-guanylate-specific phosphodiesterase activity | IBA GO_REF:0000033 | ACCEPT | Summary: IBA from the 18-seed PTN002105365 IBD; PA2133 appears among its own sources because of its own IDA row, which is expected. Reason: Correct for PA2133 despite the ETL motif: lysate and in vivo data show active c-di-GMP hydrolysis. Node-level defects are recorded in the rocR review. Supporting Evidence: PMID:16186483 wspF mutant cells carrying the PA2133 expression vector had undetectable levels of c-diGMP |
| GO:1900190 regulation of single-species biofilm formation | IBA GO_REF:0000033 | KEEP AS NON CORE | Summary: IBA from the 4-seed PTN009140332 IBD. PA2133's own seed is an overexpression-as-tool phenotype, but the IBD stands on bifA's deletion-plus-complementation evidence. Reason: A c-di-GMP PDE this active can plausibly regulate biofilm formation, and the node is sound, but native PA2133 has no loss-of-function biofilm phenotype. Kept as an inherited, non-core process. Supporting Evidence: PMID:16186483 A PA2133 mutant resembled the wild type in attachment assays. |
| GO:0071111 cyclic-guanylate-specific phosphodiesterase activity | IDA PMID:16477007 Analysis of Pseudomonas aeruginosa diguanylate cyclases and ... | ACCEPT | Summary: PseudoCAP IDA and PA2133's seeding of PTN002105365. Lysates of a PA2133-overexpressing strain degraded 71% of added c-di-GMP (Table 1 and Fig. 3B, full text read on PMC). Reason: Seed verdict SOUND. A lysate assay is crude but direct, and it agrees with the in vivo depletion of c-di-GMP by PA2133. Supporting Evidence: file:PSEAE/PA2133/PA2133-notes.md PA2133 ETL — +(71) PMID:16186483 wspF mutant cells carrying the PA2133 expression vector had undetectable levels of c-diGMP |
| GO:1900191 negative regulation of single-species biofilm formation | IMP PMID:16186483 A chemosensory system that regulates biofilm formation throu... | MARK AS OVER ANNOTATED | Summary: PseudoCAP IMP and PA2133's seeding of PTN009140332. In this paper PA2133 is expressed from a plasmid as a tool to drain c-di-GMP, which blocks attachment and biofilm. The PA2133 mutant resembles wild type. Reason: Seed verdict INDIRECT-PHENOTYPE (full text read). The phenotype is that of artificial c-di-GMP depletion. It shows PA2133 is an effective PDE, not that native PA2133 negatively regulates biofilm formation. The term is not implausible, so the row is marked over-annotated rather than removed. The PTN009140332 IBD does not depend on this seed. Supporting Evidence: PMID:16186483 A PA2133 mutant resembled the wild type in attachment assays. PMID:16186483 the ability of wild-type cells to attach to surfaces was also completely abolished when PA2133 was expressed |
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Download this section (compressed HTML)Experiment: Profile PA2133 expression across growth and biofilm conditions with a chromosomal reporter, and test a clean PA2133 deletion for biofilm, dispersal, motility and cytotoxicity phenotypes under the conditions where it is expressed.
Experiment: Purify PA2133 and measure c-di-GMP hydrolysis kinetics to test whether the ETL motif tunes activity compared with canonical EAL PDEs.
What is not known — curated, literature-grounded statements of the open unknowns (the inverse of core functions).
Gap: The physiological role of native PA2133 is unknown: when and where it is expressed, which c-di-GMP pool it controls, and which output it regulates. Its loss-of-function mutant has no attachment phenotype.
BIOLOGY
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