{"gene":"ANAPC11","run_date":"2026-06-09T22:02:43","timeline":{"discoveries":[{"year":1999,"finding":"APC11 (and its homologs ROC1/ROC2) immunocomplexes can catalyze isopeptide ligations to form polyubiquitin chains in an E1- and E2-dependent manner; APC11 specifically interacts with APC2 (a cullin-related APC subunit), while ROC1/ROC2 interact with all cullins; ROC1/APC11 RING finger mutations completely abolished ligase activity without disrupting associated protein interactions.","method":"In vitro ubiquitin ligase assay with immunocomplexes; mutagenesis of RING finger; co-immunoprecipitation","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution of ligase activity, mutagenesis abolishing activity, and binding specificity established; replicated across multiple constructs","pmids":["10230407"],"is_preprint":false},{"year":2000,"finding":"APC11 alone is sufficient to synthesize multiubiquitin chains in the presence of E1 and UBC4; APC11 and UBC4 together can ubiquitinate securin and cyclin B with reduced D-box dependency; the RING-H2 finger domain is required for ubiquitination activity.","method":"In vitro ubiquitination assay with recombinant E. coli-expressed proteins; RING finger mutagenesis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with purified recombinant proteins plus mutagenesis; replicated by multiple labs","pmids":["10922056"],"is_preprint":false},{"year":2000,"finding":"Yeast Apc11p RING-H2 finger is essential for cell viability; Apc11p directly interacts with the Ubc4 E2 enzyme; purified Apc11p mediates E1- and E2-dependent ubiquitination of substrates including Clb2p in vitro; E3 activity requires RING-H2 finger integrity but does not require the cullin-like Apc2p subunit.","method":"Genetic viability assay (yeast); direct binding assay with purified recombinant proteins; in vitro ubiquitination assay; RING-H2 mutagenesis","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1 / Strong — reconstitution with purified proteins, mutagenesis, and genetic essentiality all established in one study; consistent with parallel papers","pmids":["10888670"],"is_preprint":false},{"year":2001,"finding":"A heterodimeric complex of APC2 and APC11 is the minimal ubiquitin ligase module of the human APC/C, sufficient to ubiquitinate securin and cyclin B1 with Ubc4 or UbcH10; APC11 and UbcH10 both bind the C-terminal cullin homology domain of APC2, while Ubc4 interacts with APC11 directly; APC11 binds Zn2+ at a 1:3 molar ratio and the third Zn2+ ion is not essential for ligase activity; with Ubc4, Zn2+ ions alone can catalyze ubiquitination of cyclin B1.","method":"Baculoviral reconstitution of human APC2/APC11 complex; in vitro ubiquitination assay; Zn2+-binding experiments; mutagenesis; binding domain mapping","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1 / Strong — reconstitution of minimal ligase module, mutagenesis, Zn2+-binding characterization, and domain mapping all in one study","pmids":["11739784"],"is_preprint":false},{"year":2001,"finding":"Human ANAPC11 protein is distributed diffusely in the cytoplasm and nucleus with discrete accumulation in granular structures, as determined by transfection-based localization experiments in AML 12, HepG2, and C2C12 cell lines.","method":"Transfection and subcellular localization imaging","journal":"Journal of cellular biochemistry","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, single method (transfection/imaging), no functional consequence linked to localization","pmids":["11573242"],"is_preprint":false},{"year":2004,"finding":"Exposure of purified APC11 to H2O2 (0.1–1 mM) releases bound zinc via oxidation of cysteine residues, impairs physical interaction between APC11 and Ubc4, and inhibits APC11-mediated ubiquitination of cyclin B1; in HeLa cells, exogenous H2O2 blocked co-immunoprecipitation of Ubc4 with APC11, inhibited cyclin B1 and securin ubiquitination and degradation, and delayed mitotic exit.","method":"In vitro ubiquitination assay with purified proteins; zinc release assay; co-immunoprecipitation in H2O2-treated HeLa cells; mitotic exit timing assay","journal":"Free radical biology & medicine","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — in vitro biochemical assays with purified protein plus cellular co-IP and functional mitotic exit assay; multiple orthogonal methods in one study","pmids":["15256223"],"is_preprint":false},{"year":2006,"finding":"Recombinant goldfish APC11 possesses ubiquitinating activity against cyclin B; a reconstituted in vitro system comprising purified E1, E2-C (UBC4 type), and APC11 is sufficient to ubiquitinate cyclin B.","method":"In vitro ubiquitination assay with recombinant proteins from goldfish (ortholog)","journal":"Zoological science","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — in vitro reconstitution with recombinant proteins; single lab, single study in non-mammalian ortholog","pmids":["16971785"],"is_preprint":false},{"year":2012,"finding":"Drosophila Apc11 (encoded by lemmingA) is essential for mitotic progression; loss-of-function causes metaphase arrest with condensed scattered chromosomes and polyploidy, and accumulation of cyclin A and cyclin B; LmgA/Apc11 interacts with Morula/Apc2 and together they form a binding site for Vihar (E2-C type ubiquitin-conjugating enzyme), establishing a ternary complex.","method":"Drosophila genetic null allele rescue; yeast complementation; co-immunoprecipitation/interaction assays; immunofluorescence","journal":"Cell division","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic null allele with defined mitotic phenotype, cross-species complementation, and physical interaction mapping; multiple orthogonal methods","pmids":["22417125"],"is_preprint":false},{"year":2012,"finding":"siRNA knockdown of Apc11 in HEK293T cells leads to reduced time in G2/M phase and increased time in G1 phase (not apoptosis); this cell cycle distribution phenotype is rescued by co-transfection of an Apc11 expression plasmid, confirming specificity.","method":"siRNA knockdown in HEK293T cells; flow cytometry cell cycle analysis; rescue experiment","journal":"Genetics and molecular research : GMR","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — clean KD with rescue control and defined cell cycle phenotype; single lab, single method","pmids":["23007976"],"is_preprint":false},{"year":1999,"finding":"Targeted disruption of APC11 (YDL008w) in S. cerevisiae is non-viable, establishing that APC11 is an essential gene.","method":"Systematic gene disruption (KanMX cassette replacement) in haploid yeast","journal":"Yeast (Chichester, England)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean genetic essentiality established by targeted disruption; replicated by other labs as noted in paper","pmids":["10487928"],"is_preprint":false},{"year":2019,"finding":"Unassembled (excess) Apc11 in yeast can ubiquitinate APC/C substrates independently of the fully assembled APC/C holoenzyme in living cells; the ubiquitin-proteasome system mediates degradation of unassembled Apc11, serving as a quality-control mechanism to prevent spurious substrate ubiquitination outside the holoenzyme context.","method":"In vivo yeast experiments with Apc11 overexpression; proteasome inhibition; substrate ubiquitination assays in living yeast","journal":"FASEB journal","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — in vivo yeast experiments with defined substrate readouts; single lab, single study","pmids":["31162950"],"is_preprint":false},{"year":2019,"finding":"APC11 mediates ubiquitination of UBA52 (ubiquitin-ribosomal fusion protein), which in turn promotes degradation of CCNB1 (cyclin B1); knockdown of APC11 causes G2/M arrest and reduced clonal formation even with UBA52 overexpression, placing APC11 upstream of UBA52 in CCNB1 degradation.","method":"Co-immunoprecipitation; shRNA/siRNA knockdown; cell cycle analysis; xenograft tumor assay; overexpression rescue experiments","journal":"American journal of translational research","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — co-IP and epistasis-like rescue experiments; multiple methods; single lab","pmids":["31814919"],"is_preprint":false},{"year":2023,"finding":"ANAPC11 interacts with and promotes ubiquitination of FOXO3, leading to decreased FOXO3 protein stability, downregulation of p21 and GULP1; CRISPR knockout of ANAPC11 inhibits tumor growth and lymph node metastasis in vivo.","method":"Co-immunoprecipitation coupled with mass spectrometry; CRISPR-Cas9 knockout; ubiquitination assay; in vivo xenograft/LN metastasis model","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — IP-MS identifies substrate, supported by ubiquitination assay and in vivo KO; single lab","pmids":["37573356"],"is_preprint":false},{"year":2024,"finding":"Cryo-EM structures of S. cerevisiae APC/C reveal that in contrast to human APC/C (where coactivator induces a conformational change of the APC2:APC11 catalytic module to permit E2 binding), in yeast apo-APC/C the APC2:APC11 catalytic module is already positioned to bind E2, indicating species-specific differences in coactivator-mediated stimulation of E2 recruitment.","method":"Cryo-EM structural determination of multiple S. cerevisiae APC/C complexes (apo, CDH1-substrate ternary, phosphorylated); comparative structural analysis with human APC/C","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — medium-resolution cryo-EM structures; preprint, not yet peer-reviewed; single study","pmids":["bio_10.1101_2024.06.19.599685"],"is_preprint":true}],"current_model":"ANAPC11 is the catalytic RING-H2 finger subunit of the APC/C E3 ubiquitin ligase: it directly recruits E2 enzymes (Ubc4/UbcH10), coordinates Zn2+ ions through its RING-H2 domain to catalyze isopeptide bond formation, forms a minimal ubiquitin ligase module with the cullin-like APC2 subunit sufficient to ubiquitinate substrates such as cyclin B and securin, and is subject to regulation by oxidative stress (H2O2-mediated Zn2+ release inhibits E2 binding and ligase activity) and by UPS-mediated degradation of its unassembled form to prevent spurious substrate ubiquitination outside the holoenzyme; additional substrates including FOXO3 and UBA52/CCNB1 have been identified in cancer contexts."},"narrative":{"mechanistic_narrative":"ANAPC11 is the catalytic RING-H2 finger subunit of the anaphase-promoting complex/cyclosome (APC/C), the multisubunit E3 ubiquitin ligase that drives ubiquitin-dependent proteolysis of cell-cycle regulators [PMID:10230407, PMID:11739784]. It functions as the catalytic core by directly binding the E2 ubiquitin-conjugating enzyme (Ubc4/UbcH10) and coordinating Zn2+ through its RING-H2 domain to catalyze isopeptide-bond formation and polyubiquitin chain synthesis; RING-H2 integrity is strictly required for ligase activity [PMID:10230407, PMID:10922056, PMID:10888670]. Together with the cullin-related subunit APC2, ANAPC11 forms the minimal heterodimeric ligase module sufficient to ubiquitinate APC/C substrates such as securin and cyclin B1, with APC11 contributing the E2-docking and chain-building activity [PMID:10922056, PMID:11739784, PMID:22417125]. This activity is required for mitotic progression: loss of ANAPC11 causes metaphase arrest, accumulation of mitotic cyclins, and altered cell-cycle distribution, and the gene is essential for viability [PMID:10888670, PMID:22417125, PMID:23007976, PMID:10487928]. Its catalytic function is regulated by oxidative stress—H2O2 oxidizes cysteine residues, releases the coordinated zinc, disrupts E2 (Ubc4) binding, and inhibits substrate ubiquitination, delaying mitotic exit [PMID:15256223]—and by ubiquitin-proteasome-mediated turnover of unassembled APC11 that prevents spurious substrate ubiquitination outside the holoenzyme [PMID:31162950]. In cancer contexts, ANAPC11 promotes ubiquitination of additional substrates including UBA52 (driving cyclin B1 degradation) and FOXO3 (destabilizing it and downregulating p21 and GULP1), with knockout suppressing tumor growth and metastasis [PMID:31814919, PMID:37573356].","teleology":[{"year":1999,"claim":"Established that APC11 is an essential gene and that its RING finger is the catalytic element of the APC/C ligase, answering whether the activity resided in a defined subunit.","evidence":"Yeast gene disruption (essentiality); in vitro ligase assay with immunocomplexes and RING-finger mutagenesis; co-IP showing APC11–APC2 interaction","pmids":["10230407","10487928"],"confidence":"High","gaps":["Did not define the minimal sufficient module","E2 specificity not yet mapped to APC11 directly"]},{"year":2000,"claim":"Showed APC11 with E1 and UBC4 is sufficient to build polyubiquitin chains and ubiquitinate securin and cyclin B, and that this requires the RING-H2 domain and direct E2 binding, pinpointing APC11 as the catalytic engine.","evidence":"In vitro ubiquitination with recombinant proteins; RING-H2 mutagenesis; direct E2 binding assay; yeast genetic essentiality","pmids":["10922056","10888670"],"confidence":"High","gaps":["Role of APC2/cullin-like partner in physiological context unresolved","Reduced D-box dependency of minimal system not reconciled with holoenzyme substrate selection"]},{"year":2001,"claim":"Defined the minimal APC2:APC11 heterodimer as the catalytic ligase module and characterized Zn2+ coordination, mapping where E2 enzymes dock and which metal ions matter for catalysis.","evidence":"Baculoviral reconstitution of human APC2/APC11; in vitro ubiquitination; Zn2+-binding stoichiometry; domain mapping","pmids":["11739784"],"confidence":"High","gaps":["Structural basis of E2 positioning not resolved","How module integrates into holoenzyme regulation unclear"]},{"year":2001,"claim":"Provided initial subcellular localization of human ANAPC11 across cell lines.","evidence":"Transfection-based localization imaging in AML 12, HepG2, C2C12","pmids":["11573242"],"confidence":"Low","gaps":["Single lab, single method with no functional link to localization","Overexpression artifact not excluded","Endogenous localization not assessed"]},{"year":2004,"claim":"Linked APC11 catalytic activity to redox regulation, showing oxidative stress disables the ligase by stripping its zinc and blocking E2 binding.","evidence":"In vitro zinc-release and ubiquitination assays with purified protein; co-IP in H2O2-treated HeLa cells; mitotic exit timing","pmids":["15256223"],"confidence":"High","gaps":["Physiological oxidant levels and reversibility in vivo not defined","Which specific cysteines are oxidized not mapped"]},{"year":2006,"claim":"Confirmed conservation of APC11 catalytic activity against cyclin B in a non-mammalian ortholog.","evidence":"In vitro ubiquitination with recombinant goldfish proteins (E1, E2-C, APC11)","pmids":["16971785"],"confidence":"Medium","gaps":["Single study in ortholog","No in vivo validation"]},{"year":2012,"claim":"Demonstrated in intact metazoan and human cells that APC11 is required for mitotic progression and forms a ternary E2-binding complex with APC2, connecting biochemistry to cell-cycle phenotype.","evidence":"Drosophila null allele rescue and complementation, co-IP, immunofluorescence; siRNA knockdown with rescue and flow cytometry in HEK293T","pmids":["22417125","23007976"],"confidence":"High","gaps":["Full substrate spectrum in vivo not enumerated","Mechanism of coactivator-dependent activation not addressed"]},{"year":2019,"claim":"Revealed a quality-control layer: unassembled APC11 retains aberrant ligase activity and is degraded by the proteasome to prevent inappropriate substrate ubiquitination.","evidence":"In vivo yeast overexpression, proteasome inhibition, substrate ubiquitination readouts","pmids":["31162950"],"confidence":"Medium","gaps":["Single lab/study","Degradation machinery targeting unassembled APC11 not identified"]},{"year":2019,"claim":"Identified UBA52 as an APC11 ubiquitination substrate that drives cyclin B1 degradation, extending APC11 function into a cancer-relevant degradation pathway.","evidence":"Co-IP, shRNA/siRNA knockdown, cell-cycle analysis, xenograft, overexpression rescue/epistasis","pmids":["31814919"],"confidence":"Medium","gaps":["Direct vs indirect ubiquitination not fully separated","Single lab"]},{"year":2023,"claim":"Established FOXO3 as an ANAPC11 substrate whose destabilization links the ligase to tumor growth and metastasis via p21/GULP1 downregulation.","evidence":"Co-IP/mass spectrometry, CRISPR-Cas9 knockout, ubiquitination assay, in vivo xenograft and lymph node metastasis model","pmids":["37573356"],"confidence":"Medium","gaps":["Whether ubiquitination occurs via the APC/C holoenzyme or APC11 alone unclear","Single lab"]},{"year":2024,"claim":"Resolved species-specific differences in catalytic module activation, showing yeast apo-APC/C pre-positions the APC2:APC11 module for E2 binding whereas human APC/C requires coactivator-induced conformational change.","evidence":"Cryo-EM of multiple S. cerevisiae APC/C complexes with comparative analysis (preprint)","pmids":["bio_10.1101_2024.06.19.599685"],"confidence":"Medium","gaps":["Preprint, not peer-reviewed","Medium resolution","Functional consequence of pre-positioning not tested biochemically"]},{"year":null,"claim":"How ANAPC11 substrate selection is partitioned between its action within the holoenzyme versus its intrinsic minimal-module activity, and how redox/coactivator regulation integrates in physiological human cells, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No high-resolution human APC2:APC11–E2–substrate structure in the corpus","Endogenous human substrate spectrum incompletely defined","In vivo significance of oxidative regulation not quantified"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[0,1,2,3]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[1,3,11,12]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[3,7]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[4]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[4]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[2,7,8,9]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,1,3,10]}],"complexes":["APC/C","APC2:APC11 catalytic module"],"partners":["APC2","UBC4","UBE2C","FOXO3","UBA52"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NYG5","full_name":"Anaphase-promoting complex subunit 11","aliases":["Cyclosome subunit 11","Hepatocellular carcinoma-associated RING finger protein"],"length_aa":84,"mass_kda":9.8,"function":"Together with the cullin protein ANAPC2, constitutes the catalytic component of the anaphase promoting complex/cyclosome (APC/C), a cell cycle-regulated E3 ubiquitin ligase that controls progression through mitosis and the G1 phase of the cell cycle (PubMed:11739784, PubMed:18485873). The APC/C complex acts by mediating ubiquitination and subsequent degradation of target proteins: it mainly mediates the formation of 'Lys-11'-linked polyubiquitin chains and, to a lower extent, the formation of 'Lys-48'- and 'Lys-63'-linked polyubiquitin chains (PubMed:11739784, PubMed:18485873). The APC/C complex catalyzes assembly of branched 'Lys-11'-/'Lys-48'-linked branched ubiquitin chains on target proteins (PubMed:29033132). May recruit the E2 ubiquitin-conjugating enzymes to the complex (PubMed:11739784, PubMed:18485873)","subcellular_location":"Cytoplasm; Nucleus","url":"https://www.uniprot.org/uniprotkb/Q9NYG5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/ANAPC11","classification":"Common Essential","n_dependent_lines":1204,"n_total_lines":1208,"dependency_fraction":0.9966887417218543},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000141552","cell_line_id":"CID000219","localizations":[{"compartment":"cytoplasmic","grade":3},{"compartment":"nucleoplasm","grade":3}],"interactors":[{"gene":"SLC7A6","stoichiometry":10.0},{"gene":"CCT7","stoichiometry":4.0},{"gene":"ANAPC4","stoichiometry":4.0},{"gene":"MAP2K7","stoichiometry":4.0},{"gene":"ANAPC2","stoichiometry":0.2},{"gene":"CSNK2A2","stoichiometry":0.2},{"gene":"SLC35B1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/target/CID000219","total_profiled":1310},"omim":[{"mim_id":"614534","title":"ANAPHASE-PROMOTING COMPLEX SUBUNIT 11; ANAPC11","url":"https://www.omim.org/entry/614534"},{"mim_id":"603863","title":"RING FINGER PROTEIN 7; RNF7","url":"https://www.omim.org/entry/603863"},{"mim_id":"603814","title":"RING-BOX 1; RBX1","url":"https://www.omim.org/entry/603814"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Nucleoplasm","reliability":"Enhanced"},{"location":"Nucleoli","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/ANAPC11"},"hgnc":{"alias_symbol":["HSPC214","APC11","Apc11p","MGC882"],"prev_symbol":[]},"alphafold":{"accession":"Q9NYG5","domains":[{"cath_id":"3.30.40.10","chopping":"16-84","consensus_level":"medium","plddt":93.6148,"start":16,"end":84}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NYG5","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NYG5-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NYG5-F1-predicted_aligned_error_v6.png","plddt_mean":92.38},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ANAPC11","jax_strain_url":"https://www.jax.org/strain/search?query=ANAPC11"},"sequence":{"accession":"Q9NYG5","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NYG5.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NYG5/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NYG5"}},"corpus_meta":[{"pmid":"10230407","id":"PMC_10230407","title":"ROC1, a homolog of APC11, represents a family of cullin partners with an associated ubiquitin ligase activity.","date":"1999","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/10230407","citation_count":410,"is_preprint":false},{"pmid":"11739784","id":"PMC_11739784","title":"APC2 Cullin protein and APC11 RING protein comprise the minimal ubiquitin ligase module of the anaphase-promoting complex.","date":"2001","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/11739784","citation_count":156,"is_preprint":false},{"pmid":"10922056","id":"PMC_10922056","title":"The RING-H2 finger protein APC11 and the E2 enzyme UBC4 are sufficient to ubiquitinate substrates of the anaphase-promoting complex.","date":"2000","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/10922056","citation_count":156,"is_preprint":false},{"pmid":"10888670","id":"PMC_10888670","title":"The APC11 RING-H2 finger mediates E2-dependent ubiquitination.","date":"2000","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/10888670","citation_count":151,"is_preprint":false},{"pmid":"31814919","id":"PMC_31814919","title":"Degradation of CCNB1 mediated by APC11 through UBA52 ubiquitination promotes cell cycle progression and proliferation of non-small cell lung cancer cells.","date":"2019","source":"American journal of translational research","url":"https://pubmed.ncbi.nlm.nih.gov/31814919","citation_count":39,"is_preprint":false},{"pmid":"15256223","id":"PMC_15256223","title":"The RING-H2-finger protein APC11 as a target of hydrogen peroxide.","date":"2004","source":"Free radical biology & medicine","url":"https://pubmed.ncbi.nlm.nih.gov/15256223","citation_count":28,"is_preprint":false},{"pmid":"32319544","id":"PMC_32319544","title":"LncRNA BCAR4 promotes liver cancer progression by upregulating ANAPC11 expression through sponging miR‑1261.","date":"2020","source":"International journal of molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/32319544","citation_count":19,"is_preprint":false},{"pmid":"37573356","id":"PMC_37573356","title":"The APC/C E3 ligase subunit ANAPC11 mediates FOXO3 protein degradation to promote cell proliferation and lymph node metastasis in urothelial bladder cancer.","date":"2023","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/37573356","citation_count":15,"is_preprint":false},{"pmid":"23007976","id":"PMC_23007976","title":"Knockdown expression of Apc11 leads to cell-cycle distribution reduction in G2/M phase.","date":"2012","source":"Genetics and molecular research : GMR","url":"https://pubmed.ncbi.nlm.nih.gov/23007976","citation_count":8,"is_preprint":false},{"pmid":"22417125","id":"PMC_22417125","title":"lemmingA encodes the Apc11 subunit of the APC/C in Drosophila melanogaster that forms a ternary complex with the E2-C type ubiquitin conjugating enzyme, Vihar and Morula/Apc2.","date":"2012","source":"Cell division","url":"https://pubmed.ncbi.nlm.nih.gov/22417125","citation_count":8,"is_preprint":false},{"pmid":"10487928","id":"PMC_10487928","title":"Disruption and functional analysis of seven ORFs on chromosome IV: YDL057w, YDL012c, YDL010w, YDL009c, YDL008w (APC11), YDL005c (MED2) and YDL003w (MCD1).","date":"1999","source":"Yeast (Chichester, England)","url":"https://pubmed.ncbi.nlm.nih.gov/10487928","citation_count":6,"is_preprint":false},{"pmid":"31162950","id":"PMC_31162950","title":"Regulation of the anaphase promoting complex/cyclosome by the degradation of its unassembled catalytic subunit, Apc11.","date":"2019","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/31162950","citation_count":6,"is_preprint":false},{"pmid":"30410368","id":"PMC_30410368","title":"Overexpression of APC11 predicts worse survival in lung adenocarcinoma.","date":"2018","source":"OncoTargets and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/30410368","citation_count":5,"is_preprint":false},{"pmid":"11573242","id":"PMC_11573242","title":"Molecular cloning and characterization of a RING-H2 finger protein, ANAPC11, the human homolog of yeast Apc11p.","date":"2001","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11573242","citation_count":3,"is_preprint":false},{"pmid":"16971785","id":"PMC_16971785","title":"Molecular cloning of cDNA encoding APC11, a catalytic component of anaphase-promoting-complex (APC/C), from goldfish (Carassius auratus), and establishment of in vitro ubiquitinating system.","date":"2006","source":"Zoological science","url":"https://pubmed.ncbi.nlm.nih.gov/16971785","citation_count":1,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.06.19.599685","title":"A comparative study of the cryo-EM structures of  <i>S. cerevisiae</i>  and human anaphase-promoting complex/cyclosome (APC/C)","date":"2024-06-20","source":"bioRxiv","url":"https://doi.org/10.1101/2024.06.19.599685","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":9950,"output_tokens":3577,"usd":0.041752,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11053,"output_tokens":3650,"usd":0.073258,"stage2_stop_reason":"end_turn"},"total_usd":0.11501,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1999,\n      \"finding\": \"APC11 (and its homologs ROC1/ROC2) immunocomplexes can catalyze isopeptide ligations to form polyubiquitin chains in an E1- and E2-dependent manner; APC11 specifically interacts with APC2 (a cullin-related APC subunit), while ROC1/ROC2 interact with all cullins; ROC1/APC11 RING finger mutations completely abolished ligase activity without disrupting associated protein interactions.\",\n      \"method\": \"In vitro ubiquitin ligase assay with immunocomplexes; mutagenesis of RING finger; co-immunoprecipitation\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution of ligase activity, mutagenesis abolishing activity, and binding specificity established; replicated across multiple constructs\",\n      \"pmids\": [\"10230407\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"APC11 alone is sufficient to synthesize multiubiquitin chains in the presence of E1 and UBC4; APC11 and UBC4 together can ubiquitinate securin and cyclin B with reduced D-box dependency; the RING-H2 finger domain is required for ubiquitination activity.\",\n      \"method\": \"In vitro ubiquitination assay with recombinant E. coli-expressed proteins; RING finger mutagenesis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with purified recombinant proteins plus mutagenesis; replicated by multiple labs\",\n      \"pmids\": [\"10922056\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"Yeast Apc11p RING-H2 finger is essential for cell viability; Apc11p directly interacts with the Ubc4 E2 enzyme; purified Apc11p mediates E1- and E2-dependent ubiquitination of substrates including Clb2p in vitro; E3 activity requires RING-H2 finger integrity but does not require the cullin-like Apc2p subunit.\",\n      \"method\": \"Genetic viability assay (yeast); direct binding assay with purified recombinant proteins; in vitro ubiquitination assay; RING-H2 mutagenesis\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — reconstitution with purified proteins, mutagenesis, and genetic essentiality all established in one study; consistent with parallel papers\",\n      \"pmids\": [\"10888670\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"A heterodimeric complex of APC2 and APC11 is the minimal ubiquitin ligase module of the human APC/C, sufficient to ubiquitinate securin and cyclin B1 with Ubc4 or UbcH10; APC11 and UbcH10 both bind the C-terminal cullin homology domain of APC2, while Ubc4 interacts with APC11 directly; APC11 binds Zn2+ at a 1:3 molar ratio and the third Zn2+ ion is not essential for ligase activity; with Ubc4, Zn2+ ions alone can catalyze ubiquitination of cyclin B1.\",\n      \"method\": \"Baculoviral reconstitution of human APC2/APC11 complex; in vitro ubiquitination assay; Zn2+-binding experiments; mutagenesis; binding domain mapping\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — reconstitution of minimal ligase module, mutagenesis, Zn2+-binding characterization, and domain mapping all in one study\",\n      \"pmids\": [\"11739784\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"Human ANAPC11 protein is distributed diffusely in the cytoplasm and nucleus with discrete accumulation in granular structures, as determined by transfection-based localization experiments in AML 12, HepG2, and C2C12 cell lines.\",\n      \"method\": \"Transfection and subcellular localization imaging\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, single method (transfection/imaging), no functional consequence linked to localization\",\n      \"pmids\": [\"11573242\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Exposure of purified APC11 to H2O2 (0.1–1 mM) releases bound zinc via oxidation of cysteine residues, impairs physical interaction between APC11 and Ubc4, and inhibits APC11-mediated ubiquitination of cyclin B1; in HeLa cells, exogenous H2O2 blocked co-immunoprecipitation of Ubc4 with APC11, inhibited cyclin B1 and securin ubiquitination and degradation, and delayed mitotic exit.\",\n      \"method\": \"In vitro ubiquitination assay with purified proteins; zinc release assay; co-immunoprecipitation in H2O2-treated HeLa cells; mitotic exit timing assay\",\n      \"journal\": \"Free radical biology & medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — in vitro biochemical assays with purified protein plus cellular co-IP and functional mitotic exit assay; multiple orthogonal methods in one study\",\n      \"pmids\": [\"15256223\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Recombinant goldfish APC11 possesses ubiquitinating activity against cyclin B; a reconstituted in vitro system comprising purified E1, E2-C (UBC4 type), and APC11 is sufficient to ubiquitinate cyclin B.\",\n      \"method\": \"In vitro ubiquitination assay with recombinant proteins from goldfish (ortholog)\",\n      \"journal\": \"Zoological science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — in vitro reconstitution with recombinant proteins; single lab, single study in non-mammalian ortholog\",\n      \"pmids\": [\"16971785\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Drosophila Apc11 (encoded by lemmingA) is essential for mitotic progression; loss-of-function causes metaphase arrest with condensed scattered chromosomes and polyploidy, and accumulation of cyclin A and cyclin B; LmgA/Apc11 interacts with Morula/Apc2 and together they form a binding site for Vihar (E2-C type ubiquitin-conjugating enzyme), establishing a ternary complex.\",\n      \"method\": \"Drosophila genetic null allele rescue; yeast complementation; co-immunoprecipitation/interaction assays; immunofluorescence\",\n      \"journal\": \"Cell division\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic null allele with defined mitotic phenotype, cross-species complementation, and physical interaction mapping; multiple orthogonal methods\",\n      \"pmids\": [\"22417125\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"siRNA knockdown of Apc11 in HEK293T cells leads to reduced time in G2/M phase and increased time in G1 phase (not apoptosis); this cell cycle distribution phenotype is rescued by co-transfection of an Apc11 expression plasmid, confirming specificity.\",\n      \"method\": \"siRNA knockdown in HEK293T cells; flow cytometry cell cycle analysis; rescue experiment\",\n      \"journal\": \"Genetics and molecular research : GMR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — clean KD with rescue control and defined cell cycle phenotype; single lab, single method\",\n      \"pmids\": [\"23007976\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"Targeted disruption of APC11 (YDL008w) in S. cerevisiae is non-viable, establishing that APC11 is an essential gene.\",\n      \"method\": \"Systematic gene disruption (KanMX cassette replacement) in haploid yeast\",\n      \"journal\": \"Yeast (Chichester, England)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean genetic essentiality established by targeted disruption; replicated by other labs as noted in paper\",\n      \"pmids\": [\"10487928\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Unassembled (excess) Apc11 in yeast can ubiquitinate APC/C substrates independently of the fully assembled APC/C holoenzyme in living cells; the ubiquitin-proteasome system mediates degradation of unassembled Apc11, serving as a quality-control mechanism to prevent spurious substrate ubiquitination outside the holoenzyme context.\",\n      \"method\": \"In vivo yeast experiments with Apc11 overexpression; proteasome inhibition; substrate ubiquitination assays in living yeast\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — in vivo yeast experiments with defined substrate readouts; single lab, single study\",\n      \"pmids\": [\"31162950\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"APC11 mediates ubiquitination of UBA52 (ubiquitin-ribosomal fusion protein), which in turn promotes degradation of CCNB1 (cyclin B1); knockdown of APC11 causes G2/M arrest and reduced clonal formation even with UBA52 overexpression, placing APC11 upstream of UBA52 in CCNB1 degradation.\",\n      \"method\": \"Co-immunoprecipitation; shRNA/siRNA knockdown; cell cycle analysis; xenograft tumor assay; overexpression rescue experiments\",\n      \"journal\": \"American journal of translational research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — co-IP and epistasis-like rescue experiments; multiple methods; single lab\",\n      \"pmids\": [\"31814919\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ANAPC11 interacts with and promotes ubiquitination of FOXO3, leading to decreased FOXO3 protein stability, downregulation of p21 and GULP1; CRISPR knockout of ANAPC11 inhibits tumor growth and lymph node metastasis in vivo.\",\n      \"method\": \"Co-immunoprecipitation coupled with mass spectrometry; CRISPR-Cas9 knockout; ubiquitination assay; in vivo xenograft/LN metastasis model\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — IP-MS identifies substrate, supported by ubiquitination assay and in vivo KO; single lab\",\n      \"pmids\": [\"37573356\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Cryo-EM structures of S. cerevisiae APC/C reveal that in contrast to human APC/C (where coactivator induces a conformational change of the APC2:APC11 catalytic module to permit E2 binding), in yeast apo-APC/C the APC2:APC11 catalytic module is already positioned to bind E2, indicating species-specific differences in coactivator-mediated stimulation of E2 recruitment.\",\n      \"method\": \"Cryo-EM structural determination of multiple S. cerevisiae APC/C complexes (apo, CDH1-substrate ternary, phosphorylated); comparative structural analysis with human APC/C\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — medium-resolution cryo-EM structures; preprint, not yet peer-reviewed; single study\",\n      \"pmids\": [\"bio_10.1101_2024.06.19.599685\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"ANAPC11 is the catalytic RING-H2 finger subunit of the APC/C E3 ubiquitin ligase: it directly recruits E2 enzymes (Ubc4/UbcH10), coordinates Zn2+ ions through its RING-H2 domain to catalyze isopeptide bond formation, forms a minimal ubiquitin ligase module with the cullin-like APC2 subunit sufficient to ubiquitinate substrates such as cyclin B and securin, and is subject to regulation by oxidative stress (H2O2-mediated Zn2+ release inhibits E2 binding and ligase activity) and by UPS-mediated degradation of its unassembled form to prevent spurious substrate ubiquitination outside the holoenzyme; additional substrates including FOXO3 and UBA52/CCNB1 have been identified in cancer contexts.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ANAPC11 is the catalytic RING-H2 finger subunit of the anaphase-promoting complex/cyclosome (APC/C), the multisubunit E3 ubiquitin ligase that drives ubiquitin-dependent proteolysis of cell-cycle regulators [#0, #3]. It functions as the catalytic core by directly binding the E2 ubiquitin-conjugating enzyme (Ubc4/UbcH10) and coordinating Zn2+ through its RING-H2 domain to catalyze isopeptide-bond formation and polyubiquitin chain synthesis; RING-H2 integrity is strictly required for ligase activity [#0, #1, #2]. Together with the cullin-related subunit APC2, ANAPC11 forms the minimal heterodimeric ligase module sufficient to ubiquitinate APC/C substrates such as securin and cyclin B1, with APC11 contributing the E2-docking and chain-building activity [#1, #3, #7]. This activity is required for mitotic progression: loss of ANAPC11 causes metaphase arrest, accumulation of mitotic cyclins, and altered cell-cycle distribution, and the gene is essential for viability [#2, #7, #8, #9]. Its catalytic function is regulated by oxidative stress—H2O2 oxidizes cysteine residues, releases the coordinated zinc, disrupts E2 (Ubc4) binding, and inhibits substrate ubiquitination, delaying mitotic exit [#5]—and by ubiquitin-proteasome-mediated turnover of unassembled APC11 that prevents spurious substrate ubiquitination outside the holoenzyme [#10]. In cancer contexts, ANAPC11 promotes ubiquitination of additional substrates including UBA52 (driving cyclin B1 degradation) and FOXO3 (destabilizing it and downregulating p21 and GULP1), with knockout suppressing tumor growth and metastasis [#11, #12].\",\n  \"teleology\": [\n    {\n      \"year\": 1999,\n      \"claim\": \"Established that APC11 is an essential gene and that its RING finger is the catalytic element of the APC/C ligase, answering whether the activity resided in a defined subunit.\",\n      \"evidence\": \"Yeast gene disruption (essentiality); in vitro ligase assay with immunocomplexes and RING-finger mutagenesis; co-IP showing APC11–APC2 interaction\",\n      \"pmids\": [\"10230407\", \"10487928\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not define the minimal sufficient module\", \"E2 specificity not yet mapped to APC11 directly\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Showed APC11 with E1 and UBC4 is sufficient to build polyubiquitin chains and ubiquitinate securin and cyclin B, and that this requires the RING-H2 domain and direct E2 binding, pinpointing APC11 as the catalytic engine.\",\n      \"evidence\": \"In vitro ubiquitination with recombinant proteins; RING-H2 mutagenesis; direct E2 binding assay; yeast genetic essentiality\",\n      \"pmids\": [\"10922056\", \"10888670\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Role of APC2/cullin-like partner in physiological context unresolved\", \"Reduced D-box dependency of minimal system not reconciled with holoenzyme substrate selection\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Defined the minimal APC2:APC11 heterodimer as the catalytic ligase module and characterized Zn2+ coordination, mapping where E2 enzymes dock and which metal ions matter for catalysis.\",\n      \"evidence\": \"Baculoviral reconstitution of human APC2/APC11; in vitro ubiquitination; Zn2+-binding stoichiometry; domain mapping\",\n      \"pmids\": [\"11739784\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of E2 positioning not resolved\", \"How module integrates into holoenzyme regulation unclear\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Provided initial subcellular localization of human ANAPC11 across cell lines.\",\n      \"evidence\": \"Transfection-based localization imaging in AML 12, HepG2, C2C12\",\n      \"pmids\": [\"11573242\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single lab, single method with no functional link to localization\", \"Overexpression artifact not excluded\", \"Endogenous localization not assessed\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Linked APC11 catalytic activity to redox regulation, showing oxidative stress disables the ligase by stripping its zinc and blocking E2 binding.\",\n      \"evidence\": \"In vitro zinc-release and ubiquitination assays with purified protein; co-IP in H2O2-treated HeLa cells; mitotic exit timing\",\n      \"pmids\": [\"15256223\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological oxidant levels and reversibility in vivo not defined\", \"Which specific cysteines are oxidized not mapped\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Confirmed conservation of APC11 catalytic activity against cyclin B in a non-mammalian ortholog.\",\n      \"evidence\": \"In vitro ubiquitination with recombinant goldfish proteins (E1, E2-C, APC11)\",\n      \"pmids\": [\"16971785\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single study in ortholog\", \"No in vivo validation\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Demonstrated in intact metazoan and human cells that APC11 is required for mitotic progression and forms a ternary E2-binding complex with APC2, connecting biochemistry to cell-cycle phenotype.\",\n      \"evidence\": \"Drosophila null allele rescue and complementation, co-IP, immunofluorescence; siRNA knockdown with rescue and flow cytometry in HEK293T\",\n      \"pmids\": [\"22417125\", \"23007976\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Full substrate spectrum in vivo not enumerated\", \"Mechanism of coactivator-dependent activation not addressed\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Revealed a quality-control layer: unassembled APC11 retains aberrant ligase activity and is degraded by the proteasome to prevent inappropriate substrate ubiquitination.\",\n      \"evidence\": \"In vivo yeast overexpression, proteasome inhibition, substrate ubiquitination readouts\",\n      \"pmids\": [\"31162950\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab/study\", \"Degradation machinery targeting unassembled APC11 not identified\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified UBA52 as an APC11 ubiquitination substrate that drives cyclin B1 degradation, extending APC11 function into a cancer-relevant degradation pathway.\",\n      \"evidence\": \"Co-IP, shRNA/siRNA knockdown, cell-cycle analysis, xenograft, overexpression rescue/epistasis\",\n      \"pmids\": [\"31814919\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect ubiquitination not fully separated\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Established FOXO3 as an ANAPC11 substrate whose destabilization links the ligase to tumor growth and metastasis via p21/GULP1 downregulation.\",\n      \"evidence\": \"Co-IP/mass spectrometry, CRISPR-Cas9 knockout, ubiquitination assay, in vivo xenograft and lymph node metastasis model\",\n      \"pmids\": [\"37573356\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether ubiquitination occurs via the APC/C holoenzyme or APC11 alone unclear\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Resolved species-specific differences in catalytic module activation, showing yeast apo-APC/C pre-positions the APC2:APC11 module for E2 binding whereas human APC/C requires coactivator-induced conformational change.\",\n      \"evidence\": \"Cryo-EM of multiple S. cerevisiae APC/C complexes with comparative analysis (preprint)\",\n      \"pmids\": [\"bio_10.1101_2024.06.19.599685\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint, not peer-reviewed\", \"Medium resolution\", \"Functional consequence of pre-positioning not tested biochemically\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How ANAPC11 substrate selection is partitioned between its action within the holoenzyme versus its intrinsic minimal-module activity, and how redox/coactivator regulation integrates in physiological human cells, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No high-resolution human APC2:APC11–E2–substrate structure in the corpus\", \"Endogenous human substrate spectrum incompletely defined\", \"In vivo significance of oxidative regulation not quantified\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [0, 1, 2, 3]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1, 3, 11, 12]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [3, 7]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [2, 7, 8, 9]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 1, 3, 10]}\n    ],\n    \"complexes\": [\"APC/C\", \"APC2:APC11 catalytic module\"],\n    \"partners\": [\"APC2\", \"UBC4\", \"UBE2C\", \"FOXO3\", \"UBA52\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}