CFL2

Summary: CFL2 directly severs actin filaments.

Reason: The ADF-homology mapping captures genuine actin binding. Independent experiments with full-length human CFL2 measure single-filament severing and define its actin-contacting conformation, supporting the more informative molecular activity GO:0003789. The original InterPro source is retained; this refinement is grounded in human protein assays, not inferred solely from the domain name.

Proposed replacements: actin filament severing activity

Summary: The source-specific association is actin binding.

Reason: Human CFL2 and ACTB were examined by yeast two-hybrid and tagged pull-down in HEK293T. Exact records intact:EBI-760952, intact:EBI-752938 link CFL2 to UniProtKB:P60709. Actin binding describes the demonstrated partner class; these screens do not resolve monomeric versus filamentous actin or measure severing. The curated records share source provenance with GOA and are not an independent replication.

Proposed replacements: actin binding

Summary: The source-specific association is actin binding.

Reason: Human CFL2 and ACTG1 were examined by yeast two-hybrid and tagged pull-down in HEK293T. Exact records intact:EBI-760924, intact:EBI-753316 link CFL2 to UniProtKB:P63261. Actin binding describes the demonstrated partner class; these screens do not resolve monomeric versus filamentous actin or measure severing. The curated records share source provenance with GOA and are not an independent replication.

Proposed replacements: actin binding

Summary: The exact 2011 screen pair remains unadjudicated.

Reason: The genuine full primary text describes the human ORFeome yeast two-hybrid screen, but the exact CFL2–CFL1 entry lies in Supplementary Table 2, which was not recovered after bounded attempts. The current IntAct query yielded no matching source pair; absence from that query is not negative biological evidence. Preserve the original IPI assertion without claiming that a different publication verifies this particular experiment.

Summary: The exact 2011 screen pair remains unadjudicated.

Reason: The genuine full primary text describes the human ORFeome yeast two-hybrid screen, but the exact CFL2–ACTB entry lies in Supplementary Table 2, which was not recovered after bounded attempts. The current IntAct query yielded no matching source pair; absence from that query is not negative biological evidence. Preserve the original IPI assertion without claiming that a different publication verifies this particular experiment.

Summary: CFL2 binds the full-length muscle LIM protein isoform.

Reason: The full primary Results and Figure 6 distinguish CSRP3/MLP-a from the short MLP-b isoform. MLP-a binds CFL2; MLP-b does not bind CFL2 directly and modulates the MLP-a–CFL2 system through MLP-a. Retain the actual CSRP3 association under the explicit project binding instruction, without assigning transcriptional or adaptor activity to CFL2 or transferring the short-isoform negative result to the full-length protein.

Summary: The source-specific association is actin binding.

Reason: The records include the wild-type human ACTB pair and ACTB mutation comparisons; residue 183 has no effect and residue 364 disrupts the interaction in this source. Exact records intact:EBI-12727132, intact:EBI-12700770, intact:EBI-12712872 link CFL2 to UniProtKB:P60709. Actin binding describes the demonstrated partner class; these screens do not resolve monomeric versus filamentous actin or measure severing. The curated records share source provenance with GOA and are not an independent replication.

Proposed replacements: actin binding

Summary: The source-specific association is actin binding.

Reason: The wild-type human ACTG1 pair and variant comparisons were examined by yeast assays and split-luciferase complementation in HEK293T; ACTG1 mutations can alter binding. Exact records intact:EBI-12727152, intact:EBI-12700782, intact:EBI-12712892, intact:EBI-12689479 link CFL2 to UniProtKB:P63261. Actin binding describes the demonstrated partner class; these screens do not resolve monomeric versus filamentous actin or measure severing. The curated records share source provenance with GOA and are not an independent replication.

Proposed replacements: actin binding

Summary: The source-specific association is actin binding.

Reason: The exact human pair was tested by yeast two-hybrid with an ACTB residue-364 variant annotated as having no effect in this source. This is not independent proof that every ACTB variant behaves identically. Exact records intact:EBI-22122069 link CFL2 to UniProtKB:P60709. Actin binding describes the demonstrated partner class; these screens do not resolve monomeric versus filamentous actin or measure severing. The curated records share source provenance with GOA and are not an independent replication.

Proposed replacements: actin binding

Summary: The source-specific association is actin binding.

Reason: The human ACTB pair is supported by the HuRI yeast two-hybrid records, including its validated interaction. Exact records intact:EBI-23140504, intact:EBI-24485446, intact:EBI-22690676 link CFL2 to UniProtKB:P60709. Actin binding describes the demonstrated partner class; these screens do not resolve monomeric versus filamentous actin or measure severing. The curated records share source provenance with GOA and are not an independent replication.

Proposed replacements: actin binding

Summary: Retain the source-supported DSTN association at its measured scope.

Reason: Exact positive human-pair records intact:EBI-22741680, intact:EBI-22733825, intact:EBI-24264285 support the CFL2–DSTN association in the yeast two-hybrid system, including recorded validation. No evidence-backed destrin/cofilin-specific binding term was established by the checked ontology search. Pair association alone does not establish a stable heterodimer or a cofilin–actin rod, and DSTN binding does not itself demonstrate additional actin binding. Retain the original supported protein-binding assertion as non-core under the ClinGen instruction. This does not reject the observed association or impose a requirement for physiological replication on a binding-class term.

Summary: Refine the tested association to the verified partner-binding class.

Reason: Exact source-linked positive human yeast two-hybrid records (intact:EBI-22988258, intact:EBI-24393112, intact:EBI-24854949) support the CFL2–RAB2A association. Official partner identity and the current GO definition support small GTPase binding. No nucleotide-exchange, GTPase-activating or trafficking role is established. The assay establishes interaction capacity in the tested system, not an endogenous physiological mechanism. This binding-class refinement remains outside the core functions: the term specifies the binding partner class, not its enzymatic activity or an independently established physiological role for CFL2.

Proposed replacements: small GTPase binding

Summary: The source-specific association is actin binding.

Reason: The human ACTG1 pair is supported by the HuRI yeast two-hybrid records, including its validated interaction. Exact records intact:EBI-24434587, intact:EBI-24930059, intact:EBI-23062321 link CFL2 to UniProtKB:P63261. Actin binding describes the demonstrated partner class; these screens do not resolve monomeric versus filamentous actin or measure severing. The curated records share source provenance with GOA and are not an independent replication.

Proposed replacements: actin binding

Summary: Refine the tested association to the verified partner-binding class.

Reason: Exact source-linked positive human yeast two-hybrid records (intact:EBI-23996710, intact:EBI-23703735, intact:EBI-24679655) support the CFL2–RNF4 association. Official partner identity and the current GO definition support ubiquitin protein ligase binding. The interaction does not assign ubiquitin ligase activity to CFL2. The assay establishes interaction capacity in the tested system, not an endogenous physiological mechanism. This binding-class refinement remains outside the core functions: the term specifies the binding partner class, not its enzymatic activity or an independently established physiological role for CFL2.

Proposed replacements: ubiquitin protein ligase binding

Summary: Retain the source-supported EHHADH association at its measured scope.

Reason: Exact positive human-pair records intact:EBI-22736531, intact:EBI-24260977, intact:EBI-22730950 support the CFL2–EHHADH association in the yeast two-hybrid system, including recorded validation. No useful EHHADH-specific or hydratase-specific binding term was established by the checked ontology search. A generic enzyme-binding label would add little molecular specificity here; binding the bifunctional protein does not identify which domain is contacted, establish a catalytic role for CFL2, or demonstrate participation in peroxisomal fatty-acid metabolism. Retain the original supported protein-binding assertion as non-core under the ClinGen instruction. This does not reject the observed association or impose a requirement for physiological replication on a binding-class term.

Summary: Refine the tested association to the verified partner-binding class.

Reason: Exact source-linked positive human yeast two-hybrid records (intact:EBI-24053298, intact:EBI-23760918, intact:EBI-24712738) support the CFL2–UBE2I/UBC9 association. Official partner identity and the current GO definition support ubiquitin-like protein conjugating enzyme binding. The original Q7KZS0 accession is retained exactly. Its official record identifies the SUMO-conjugating enzyme; the assay does not demonstrate SUMO transfer by CFL2. The assay establishes interaction capacity in the tested system, not an endogenous physiological mechanism. This binding-class refinement remains outside the core functions: the term specifies the binding partner class, not its enzymatic activity or an independently established physiological role for CFL2.

Proposed replacements: ubiquitin-like protein conjugating enzyme binding

Summary: Refine the tested association to the verified partner-binding class.

Reason: Exact source-linked positive human yeast two-hybrid records (intact:EBI-22822208, intact:EBI-24306066, intact:EBI-22364363) support the CFL2–TRIM7 association. Official partner identity and the current GO definition support ubiquitin protein ligase binding. No ligase regulation or ubiquitination of CFL2 is inferred. The assay establishes interaction capacity in the tested system, not an endogenous physiological mechanism. This binding-class refinement remains outside the core functions: the term specifies the binding partner class, not its enzymatic activity or an independently established physiological role for CFL2.

Proposed replacements: ubiquitin protein ligase binding

Summary: Retain the source-supported POT1 association at its measured scope.

Reason: Exact positive human-pair records intact:EBI-24621979, intact:EBI-23603820, intact:EBI-23379443 support the CFL2–POT1 association in the yeast two-hybrid system, including recorded validation. No useful POT1-specific binding term was established by the checked ontology search. Binding one shelterin protein does not establish binding to the assembled shelterin complex or to telomeric DNA, and does not assign telomere maintenance to CFL2. Retain the original supported protein-binding assertion as non-core under the ClinGen instruction. This does not reject the observed association or impose a requirement for physiological replication on a binding-class term.

Summary: Refine the tested association to the verified partner-binding class.

Reason: Exact source-linked positive human yeast two-hybrid records (intact:EBI-26000566, intact:EBI-26154713, intact:EBI-25844037) support the CFL2–DYNC1H1 association. Official partner identity and the current GO definition support dynein heavy chain binding. No motor activity, cargo-adaptor bridge or directional transport by CFL2 is demonstrated. The assay establishes interaction capacity in the tested system, not an endogenous physiological mechanism. This binding-class refinement remains outside the core functions: the term specifies the binding partner class, not its enzymatic activity or an independently established physiological role for CFL2.

Proposed replacements: dynein heavy chain binding

Summary: Refine the positive KLK6 association to protease binding in the tested system.

Reason: Exact positive human-pair records intact:EBI-26042416, intact:EBI-26196563 and intact:EBI-25887067 support the CFL2–KLK6 association in yeast two-hybrid assays, including recorded validation. The official Q92876 identity is the serine protease KLK6, which supports protease binding under the current term definition. The records do not resolve the tested processing state or exact construct boundaries; the protein has an annotated signal peptide and activation peptide, but these cannot be assumed to have been removed in this assay. This refinement therefore describes binding to the KLK6 construct at the assay scope, not inhibition, proteolysis, a mature-protease assay or an established endogenous encounter. It remains outside the core functions, on the same definition-based criterion as the other specific partner-binding refinements.

Proposed replacements: protease binding

Summary: CFL1 and CFL2 coassociate in the BioPlex experiments.

Reason: Exact human records EBI-54768607 and EBI-54959640 link the pair to tagged co-immunoprecipitation in HEK293T and HCT116. This supports complex association, potentially involving actin, without proving a purified binary interface or an obligate cofilin heterodimer. Retain non-core under the explicit project instruction.

Summary: OpenCell supports an actin-associated CFL2 protein group.

Reason: The official interaction CSV contains ACTB as bait and the CFL2 group Q9Y281;Q9Y281-3;F8WDN3;G3V2U0 as prey. The bait library metadata identifies endogenous N-terminally tagged human ACTB. The source therefore supports actin association in the AP-MS system, corroborated by direct CFL2–actin experiments, but does not resolve each CFL2 isoform or the F-actin/G-actin state. Preserve the original source assertion and refine to actin binding.

Proposed replacements: actin binding

Summary: OpenCell supports an actin-associated CFL2 protein group.

Reason: The official interaction CSV contains ACTG1 as bait and the CFL2 group Q9Y281;Q9Y281-3;F8WDN3;G3V2U0 as prey. The bait library metadata identifies endogenous N-terminally tagged human ACTG1. The source therefore supports actin association in the AP-MS system, corroborated by direct CFL2–actin experiments, but does not resolve each CFL2 isoform or the F-actin/G-actin state. Preserve the original source assertion and refine to actin binding.

Proposed replacements: actin binding

Summary: The exact tear-proteomics target entry was not recovered.

Reason: The authentic abstract describes tear proteomics in untreated primary open-angle glaucoma and controls. Publisher supplementary files were identified but their actual CFL2 accession/peptide row could not be inspected; challenge and error responses were excluded. Preserve the HDA assertion rather than infer secretion, extracellular function or absence from a cytosolic protein annotation.

Summary: CFL2 performs its actin-regulatory work in the cytoplasm.

Reason: The PAINT assertion concerns activity in the cytoplasm and is consistent with independently measured human CFL2 actin severing and muscle sarcomeric localization. This is intracellular cytoskeletal activity, not an inference of bulk cytoplasmic localization from a membrane-bound tail. The detailed ancestral node history was not reconstructed.

Summary: CFL2 is an actin-cytoskeletal protein.

Reason: The conservative UniProt subcellular mapping agrees with direct actin-filament association and human sarcomeric localization. The broad cytoskeleton term correctly includes the actin framework in which CFL2 acts.

Summary: CFL2 participates directly in actin assembly/disassembly dynamics.

Reason: Direct human CFL2 assays establish depolymerization and severing; severing can also increase available filament ends in bulk polymerization assays. The broad ARBA process is therefore sound without claiming that CFL2 is an actin polymerase. More specific source annotations and the core describe its performed disassembly/severing work.

Summary: CFL2 acts in the actin cytoskeleton.

Reason: Direct human actin-binding/severing experiments and human muscle localization corroborate the phylogenetically inferred active-in assertion. The original qualifier and source entities are retained, without substituting donor count for assessment of the actual activity.

Summary: CFL2 acts in the actin cytoskeleton.

Reason: Direct human actin-binding/severing experiments and human muscle localization corroborate the InterPro-derived located-in assertion. The original qualifier and source entities are retained, without substituting donor count for assessment of the actual activity.

Summary: Nuclear-matrix specificity remains unresolved.

Reason: The original UniProt subcellular mapping is genuine and the selected UniProt record labels nuclear matrix by similarity. Current GO:0016363 denotes a specific nuclear protein/RNA framework; a nuclear-localization motif or broad nuclear actin-rod statement does not by itself resolve association with that framework. The historical donor experiment underlying this precise mapping was not reconstructed. This is an evidence-specific uncertainty, not a claim that CFL2 cannot enter the nucleus.

Summary: Human muscle CFL2 localizes to the Z disc.

Reason: The selected full primary study colocalizes CFL2 with the alpha-actinin Z-disc marker in human quadriceps. This supports a native muscle location associated with its actin-regulatory function; it does not make every cofilin molecule constitutively Z-disc restricted.

Summary: CFL2 directly supports actin filament depolymerization.

Reason: The PAINT inference is independently corroborated by actual human CFL2 biochemistry; the ancestral node placement itself was not reconstructed. Severing and monomer loss are related but distinct measurements; the 2009 pyrene assay supplies direct depolymerization evidence.

Summary: CFL2 directly supports actin filament depolymerization.

Reason: Full-length human CFL2 expressed as a GST fusion was tested directly in pyrene-actin depolymerization assays. The kit actin species was not specified in the inspected Methods and is not inferred from the human construct. Severing and monomer loss are related but distinct measurements; the 2009 pyrene assay supplies direct depolymerization evidence.

Summary: CFL2 directly supports actin filament depolymerization.

Reason: The InterPro mapping is supported by independent human protein assays, rather than accepted merely from family membership. Severing and monomer loss are related but distinct measurements; the 2009 pyrene assay supplies direct depolymerization evidence.

Summary: CFL2 directly supports actin filament depolymerization.

Reason: The literal ISS source P45591 is mouse Cfl2. Current donor GOA includes experimental depolymerization assertions; independent human biochemistry supports conservation without rewriting this curator-supplied ISS source. Severing and monomer loss are related but distinct measurements; the 2009 pyrene assay supplies direct depolymerization evidence.

Summary: CFL2 promotes actin-filament disassembly in muscle.

Reason: The original human A35T study reports reduced CFL2 and F-actin accumulation in affected muscle plus altered recombinant-protein solubility. It does not measure a purified mutant depolymerization rate. Retain the existing IMP regulatory process in that scope, independently corroborated by direct human CFL2 depolymerization; the qualifier and original source remain unchanged. This regulatory annotation overlaps the independently established execution of depolymerization; it is retained as the original broader IMP assertion, not evidence for a separate upstream regulatory mechanism.

Summary: The original human muscle study reports I-band localization.

Reason: Control human muscle staining is reported at the I band, with some localization data not shown and the acknowledged antibody cross-reaction with CFL1. Later independent human CFL2 localization resolves the Z-disc region more specifically. Preserve the original I-band assertion and its assay limitation rather than treating the two spatial descriptions as contradictory.

Summary: CFL2 itself severs actin filaments.

Reason: The PAINT process is corroborated by direct counting of severing events after adding purified full-length human CFL2 in single-filament TIRF experiments. This is performed molecular work, not only a knockout phenotype. The original PTN000227259 source remains unchanged.

Summary: CFL2 directly binds filamentous actin.

Reason: Human CFL2 assembled on actin filaments was examined structurally and biochemically. This independently corroborates the PAINT inference without reconstructing its node history.

Summary: CFL2 directly binds filamentous actin.

Reason: Human CFL2 assembled on actin filaments was examined structurally and biochemically. The preserved ISS donor P45591 is mouse Cfl2; current experimental donor annotations and direct human data agree with the conserved activity.

Summary: The exact exosome-proteomics CFL2 entry remains unverified.

Reason: The genuine primary body describes LC-MS/MS of vesicles isolated from expressed prostatic secretions in urine, but the CFL2 accession/peptide entry in the supplementary protein list was not recovered. Preserve the source HDA assertion without inferring an extracellular mechanism from a general proteome description or treating intracellular function as disproof.