{"gene":"ANAPC10","run_date":"2026-06-09T22:02:43","timeline":{"discoveries":[{"year":2010,"finding":"Cryo-EM structure of APC/C(Cdh1) bound to a D-box peptide at ~10 Å resolution revealed that Cdh1 and Apc10 together form a co-receptor for the D-box degron motif. Cdh1 repositions toward Apc10 within the central cavity of the APC/C, and NMR spectroscopy demonstrated direct D-box–Apc10 interactions, establishing that Apc10 directly contributes to D-box recognition alongside the co-activator.","method":"Single-particle cryo-EM, difference mapping, NMR spectroscopy","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 / Strong — cryo-EM structure plus NMR with direct binding validation; multiple orthogonal methods in one rigorous study","pmids":["21107322"],"is_preprint":false},{"year":2001,"finding":"Crystal structure of human APC10/DOC1 at 1.6 Å resolution revealed a beta-sandwich jellyroll fold homologous to ligand-binding domains of galactose oxidase and coagulation factor Va. Biochemical experiments further demonstrated that the C-terminus of APC10 directly binds CDC27/APC3, a TPR-repeat-containing APC subunit.","method":"X-ray crystallography (1.6 Å); biochemical binding assay (C-terminus of APC10 binding to CDC27/APC3)","journal":"Nature structural biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure plus biochemical binding assay, single lab but two orthogonal methods","pmids":["11524682"],"is_preprint":false},{"year":2002,"finding":"Crystal structure of S. cerevisiae Doc1/Apc10 at 2.2 Å resolution showed a beta-sandwich homologous to the galactose-binding domain of galactose oxidase, the C2 domain of coagulation factor, and XRCC1. Residues invariant across Doc1/Apc10 sequences, including a temperature-sensitive mitotic arrest mutant, map to a beta-sheet region proposed to mediate biomolecular interactions and APC ubiquitination function.","method":"X-ray crystallography (2.2 Å); comparative structural analysis with mutagenesis data mapping","journal":"Journal of molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure combined with functional mutant mapping, single lab","pmids":["11884135"],"is_preprint":false},{"year":1999,"finding":"Doc1/Apc10 was shown to be a stoichiometric subunit of the yeast APC throughout the cell cycle. Mutation of Doc1/Apc10 inactivates APC ubiquitination activity without destabilizing the complex. The orthologous human APC10 protein is also a genuine APC subunit in vertebrates (human and frog), and its cellular levels and APC association are not cell-cycle-regulated, as determined by biochemical fractionation and mass spectrometric analysis.","method":"Immunoprecipitation, biochemical fractionation, mass spectrometry, in vivo ubiquitination assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP, mass spectrometry, functional ubiquitination assays; replicated in yeast and vertebrates","pmids":["10318877"],"is_preprint":false},{"year":1998,"finding":"In fission yeast, apc10+ is essential for viability and required for ubiquitination and degradation of mitotic B-type cyclins. apc10 mutants show temperature-sensitive growth with defects in chromosome segregation and fail to arrest at G1 upon nitrogen starvation. A subpopulation of Apc10 co-immunoprecipitates with the APC, though it does not co-sediment with the 20S complex, suggesting a regulatory association.","method":"Genetic mutant analysis, co-immunoprecipitation, cell cycle analysis, ubiquitination assay","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic loss-of-function with defined mitotic phenotype, co-IP, ubiquitination assay in fission yeast","pmids":["9736616"],"is_preprint":false},{"year":1999,"finding":"Human APC10/Doc1 binds APC core subunits throughout the cell cycle and localizes to centrosomes and mitotic spindles during mitosis, to kinetochores from prophase to anaphase, and to the midbody during telophase/cytokinesis, as determined by co-immunoprecipitation and immunofluorescence localization studies.","method":"Co-immunoprecipitation; immunofluorescence microscopy (subcellular localization)","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — co-IP and direct localization imaging, single lab, two orthogonal methods but no functional consequence of localization directly tested","pmids":["10498862"],"is_preprint":false},{"year":2004,"finding":"ANAPC10 physically interacts with Smad3 (via the MH2 domain), and together with CDH1 forms a complex with HEF1 (NEDD9). Domain mapping showed distinct Smad3 MH2 subdomains bind APC10 and HEF1. Overexpression of APC10 and CDH1 regulated HEF1 protein levels, suggesting Smad3 recruits the APC/C to HEF1 for ubiquitination and proteasomal degradation via direct Smad3–APC10 interaction.","method":"Co-immunoprecipitation, domain mapping, overexpression/protein level assay","journal":"BMC cell biology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-IP and domain mapping, single lab, multiple interaction partners tested but no in vitro reconstitution","pmids":["15144564"],"is_preprint":false},{"year":2019,"finding":"The pseudosubstrate APC/C inhibitor Acm1 from budding yeast suppresses APC/C activity by combining high-affinity Cdh1 binding with a C-terminal D-box extension that specifically disrupts the normal interaction with Doc1/Apc10, thereby perturbing reaction processivity in ubiquitylation. Mutation of the conserved D-box converted Acm1 into an ABBA-motif-dependent APC/CCdh1 substrate, and biochemical analysis confirmed the extension's role in inhibiting processivity via Doc1/Apc10.","method":"In vivo mutant analysis, biochemical APC/C activity assays, site-directed mutagenesis","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — biochemical assay with mutagenesis and in vivo validation, single lab","pmids":["31562243"],"is_preprint":false},{"year":2001,"finding":"Disruption of the mouse Apc10/Doc1 gene underlies the oligosyndactylism (Os) radiation-induced mutation and two transgene-induced alleles (94-A and 94-K), all exhibiting a cell-autonomous block in metaphase-to-anaphase transition, establishing that Apc10/Doc1 is required for this cell cycle transition in vivo.","method":"Genetic mapping, molecular characterization of three mutant alleles, cell cycle phenotype analysis","journal":"Genomics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — three independent loss-of-function alleles with defined mitotic arrest phenotype, single study","pmids":["11247669"],"is_preprint":false},{"year":2007,"finding":"In Drosophila, loss-of-function mutations in Apc10/Doc1 cause metaphase-like arrest, chromosome overcondensation, high mitotic index, and accumulation of cyclin B (an APC/C substrate) in larval neuroblasts, establishing that Apc10/Doc1 is essential for APC/C E3 ubiquitin ligase activity and cyclin B ubiquitination in vivo.","method":"Genetic loss-of-function (mutant alleles), immunostaining for cyclin B, mitotic index quantification","journal":"Acta biologica Hungarica","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO with defined substrate accumulation phenotype, single lab","pmids":["18297794"],"is_preprint":false},{"year":2021,"finding":"During interphase, APC10 interacts with NLRP3 and promotes NLRP3 inflammasome activation; during mitosis, APC10 dissociates from NLRP3 to repress inflammatory responses, establishing a cell-cycle-dependent switch role for APC10 in innate immune signaling.","method":"Co-immunoprecipitation, cell cycle stage-specific analysis, inflammasome activation assays","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-IP plus functional inflammasome assay, single lab, two orthogonal approaches","pmids":["34407203"],"is_preprint":false},{"year":2012,"finding":"ANAPC10 protein is mainly expressed in the cytoplasm of spermatogonia and leptotene/pachytene spermatocytes in the developing mouse testis, as established by immunofluorescence and in situ hybridization.","method":"In situ hybridization, immunofluorescence microscopy","journal":"Biology of reproduction","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, localization only, no direct functional consequence demonstrated for this tissue-specific localization","pmids":["22190705"],"is_preprint":false}],"current_model":"ANAPC10 (APC10/DOC1) is a core, stoichiometric subunit of the APC/C ubiquitin ligase whose jellyroll beta-sandwich DOC domain directly contacts the destruction (D-box) degron of substrates in concert with the co-activator Cdh1, stabilizes the active holoenzyme for processive ubiquitylation, binds the APC subunit CDC27/APC3 via its C-terminus, and is essential for APC/C-dependent cyclin B ubiquitination and the metaphase-to-anaphase transition; it additionally interacts with Smad3 to recruit the APC/C for HEF1 degradation and, during interphase, binds NLRP3 to promote inflammasome activation in a cell-cycle-regulated manner."},"narrative":{"mechanistic_narrative":"ANAPC10 (APC10/DOC1) is a core, stoichiometric subunit of the anaphase-promoting complex/cyclosome (APC/C) E3 ubiquitin ligase that is required for substrate degron recognition and the metaphase-to-anaphase transition [PMID:10318877, PMID:11247669]. Its structure is a beta-sandwich jellyroll DOC domain, and its C-terminus directly binds the TPR-repeat APC subunit CDC27/APC3 to anchor it within the complex [PMID:11524682, PMID:11884135]. Functionally, APC10 together with the co-activator Cdh1 forms a co-receptor for the destruction-box (D-box) degron: cryo-EM positions Cdh1 adjacent to Apc10 in the central APC/C cavity, and NMR demonstrates direct D-box–Apc10 contacts, explaining why APC10 is needed for processive substrate ubiquitylation rather than for complex assembly [PMID:21107322, PMID:31562243]. Loss of APC10 inactivates APC/C ubiquitination activity without destabilizing the complex, causing cell-autonomous metaphase arrest and accumulation of the substrate cyclin B across yeast, fly, and mouse systems [PMID:10318877, PMID:9736616, PMID:18297794]. Beyond its constitutive APC/C role, APC10 interacts with Smad3 to recruit the APC/C–CDH1 ligase for degradation of HEF1/NEDD9 [PMID:15144564], and during interphase binds NLRP3 to promote inflammasome activation, dissociating during mitosis—a cell-cycle-gated function in innate immune signaling [PMID:34407203].","teleology":[{"year":1998,"claim":"Established that Apc10/Doc1 is essential for APC/C-mediated mitotic cyclin destruction, linking it directly to chromosome segregation control.","evidence":"Genetic loss-of-function, co-IP, and ubiquitination assays in fission yeast","pmids":["9736616"],"confidence":"High","gaps":["Mechanism of D-box recognition not yet defined","Only a subpopulation co-IPs with APC, leaving stoichiometry ambiguous at the time"]},{"year":1999,"claim":"Resolved whether Apc10 is a regulatory factor or a true subunit by showing it is a stoichiometric, cell-cycle-invariant APC component whose mutation abolishes ubiquitination without disassembling the complex.","evidence":"Reciprocal co-IP, biochemical fractionation, mass spectrometry, and in vivo ubiquitination assays in yeast and vertebrates","pmids":["10318877"],"confidence":"High","gaps":["Molecular basis of how a stable subunit contributes to catalysis unresolved","No structural model"]},{"year":1999,"claim":"Provided spatial context by mapping human APC10 to centrosomes, spindles, kinetochores, and the midbody through mitosis.","evidence":"Co-IP and immunofluorescence microscopy in human cells","pmids":["10498862"],"confidence":"Medium","gaps":["Functional consequence of each localization not tested","Single lab"]},{"year":2001,"claim":"Defined the APC10 fold as a beta-sandwich jellyroll resembling ligand-binding domains and showed its C-terminus binds CDC27/APC3, explaining how it is tethered within the APC/C.","evidence":"1.6 Å X-ray crystallography plus biochemical binding assay","pmids":["11524682"],"confidence":"High","gaps":["Did not identify the ligand engaged by the jellyroll surface","D-box binding not yet demonstrated"]},{"year":2001,"claim":"Demonstrated in mammals that Apc10/Doc1 disruption causes a cell-autonomous metaphase-to-anaphase block, confirming in vivo requirement in a vertebrate.","evidence":"Genetic mapping and characterization of three mouse mutant alleles with cell cycle phenotyping","pmids":["11247669"],"confidence":"Medium","gaps":["Substrate-level mechanism not directly assayed","Single study"]},{"year":2002,"claim":"Mapped functionally essential, invariant residues—including a temperature-sensitive arrest mutant—onto a beta-sheet surface, predicting the interaction face used for APC ubiquitination.","evidence":"2.2 Å crystallography of yeast Doc1 with mutagenesis mapping","pmids":["11884135"],"confidence":"High","gaps":["The predicted interaction partner of the beta-sheet face not yet identified structurally"]},{"year":2004,"claim":"Extended APC10 function beyond the core cell cycle by showing Smad3 directly binds APC10 to recruit the APC/C–CDH1 ligase for HEF1/NEDD9 degradation.","evidence":"Co-IP, domain mapping, and overexpression protein-level assays","pmids":["15144564"],"confidence":"Medium","gaps":["No in vitro reconstitution of HEF1 ubiquitination","Single lab"]},{"year":2010,"claim":"Settled how APC10 contributes to specificity by showing it acts with Cdh1 as a bipartite D-box co-receptor, with direct Apc10–D-box contacts within the APC/C cavity.","evidence":"Single-particle cryo-EM with difference mapping and NMR of D-box–Apc10 binding","pmids":["21107322"],"confidence":"High","gaps":["Atomic-resolution contacts limited by ~10 Å EM","Generality across diverse D-box substrates not exhaustively tested"]},{"year":2019,"claim":"Reinforced the processivity role by showing the inhibitor Acm1 uses a D-box extension to specifically disrupt the Doc1/Apc10 interaction, confirming Apc10–degron engagement drives processive ubiquitylation.","evidence":"In vivo mutant analysis, biochemical APC/C activity assays, and site-directed mutagenesis in budding yeast","pmids":["31562243"],"confidence":"Medium","gaps":["Quantitative kinetics of processivity loss not fully resolved","Single lab"]},{"year":2021,"claim":"Identified a moonlighting role in which APC10 binds NLRP3 in interphase to promote inflammasome activation and dissociates in mitosis, coupling cell cycle phase to innate immune output.","evidence":"Co-IP and cell-cycle stage-specific inflammasome activation assays","pmids":["34407203"],"confidence":"Medium","gaps":["Whether this requires APC/C catalytic activity or is APC/C-independent unresolved","Single lab, no reciprocal in vivo validation"]},{"year":null,"claim":"How APC10's non-canonical interactions (Smad3/HEF1, NLRP3) are mechanistically integrated with—or separated from—its constitutive APC/C role remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No reconstituted system distinguishing APC/C-dependent from APC/C-independent APC10 activities","Physiological significance of testis-restricted expression uncharacterized"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,3,4,9]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,1,6]}],"localization":[{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[5]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[11]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[3,4,8,9]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,3,6]}],"complexes":["APC/C (anaphase-promoting complex/cyclosome)"],"partners":["CDC27","CDH1","SMAD3","NEDD9","NLRP3"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9UM13","full_name":"Anaphase-promoting complex subunit 10","aliases":["Cyclosome subunit 10"],"length_aa":185,"mass_kda":21.3,"function":"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: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:18485873). The APC/C complex catalyzes assembly of branched 'Lys-11'-/'Lys-48'-linked branched ubiquitin chains on target proteins (PubMed:29033132)","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/Q9UM13/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/ANAPC10","classification":"Common Essential","n_dependent_lines":1188,"n_total_lines":1208,"dependency_fraction":0.9834437086092715},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"ANAPC16","stoichiometry":10.0},{"gene":"ANAPC4","stoichiometry":10.0},{"gene":"CDC16","stoichiometry":10.0},{"gene":"CDC23","stoichiometry":10.0},{"gene":"ANAPC2","stoichiometry":4.0},{"gene":"CDC26","stoichiometry":4.0},{"gene":"CDC27","stoichiometry":4.0},{"gene":"HIST2H2BE","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/ANAPC10","total_profiled":1310},"omim":[{"mim_id":"619459","title":"ZINC FINGER ZZ-TYPE DOMAIN- AND EF-HAND DOMAIN-CONTAINING PROTEIN 1; ZZEF1","url":"https://www.omim.org/entry/619459"},{"mim_id":"614534","title":"ANAPHASE-PROMOTING COMPLEX SUBUNIT 11; ANAPC11","url":"https://www.omim.org/entry/614534"},{"mim_id":"613745","title":"ANAPHASE-PROMOTING COMPLEX, SUBUNIT 10; ANAPC10","url":"https://www.omim.org/entry/613745"},{"mim_id":"609110","title":"F-BOX ONLY PROTEIN 43; FBXO43","url":"https://www.omim.org/entry/609110"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Golgi apparatus","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/ANAPC10"},"hgnc":{"alias_symbol":["APC10","DOC1","DKFZP564L0562"],"prev_symbol":[]},"alphafold":{"accession":"Q9UM13","domains":[{"cath_id":"2.60.120.260","chopping":"12-180","consensus_level":"high","plddt":91.3005,"start":12,"end":180}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9UM13","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9UM13-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9UM13-F1-predicted_aligned_error_v6.png","plddt_mean":90.19},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ANAPC10","jax_strain_url":"https://www.jax.org/strain/search?query=ANAPC10"},"sequence":{"accession":"Q9UM13","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9UM13.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9UM13/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9UM13"}},"corpus_meta":[{"pmid":"21107322","id":"PMC_21107322","title":"Structures of APC/C(Cdh1) with substrates identify Cdh1 and Apc10 as the D-box co-receptor.","date":"2010","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/21107322","citation_count":172,"is_preprint":false},{"pmid":"9736616","id":"PMC_9736616","title":"Apc10 and Ste9/Srw1, two regulators of the APC-cyclosome, as well as the CDK inhibitor Rum1 are required for G1 cell-cycle arrest in fission yeast.","date":"1998","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/9736616","citation_count":88,"is_preprint":false},{"pmid":"10318877","id":"PMC_10318877","title":"Characterization of the DOC1/APC10 subunit of the yeast and the human anaphase-promoting complex.","date":"1999","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10318877","citation_count":78,"is_preprint":false},{"pmid":"11524682","id":"PMC_11524682","title":"Crystal structure of the APC10/DOC1 subunit of the human anaphase-promoting complex.","date":"2001","source":"Nature structural biology","url":"https://pubmed.ncbi.nlm.nih.gov/11524682","citation_count":77,"is_preprint":false},{"pmid":"11884135","id":"PMC_11884135","title":"Implications for the ubiquitination reaction of the anaphase-promoting complex from the crystal structure of the Doc1/Apc10 subunit.","date":"2002","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/11884135","citation_count":42,"is_preprint":false},{"pmid":"15144564","id":"PMC_15144564","title":"Direct interaction between Smad3, APC10, CDH1 and HEF1 in proteasomal degradation of HEF1.","date":"2004","source":"BMC cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/15144564","citation_count":38,"is_preprint":false},{"pmid":"10498862","id":"PMC_10498862","title":"Identification of human APC10/Doc1 as a subunit of anaphase promoting 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potential chemopreventive agents in the Apc(Min) mouse model in vivo.","date":"2009","source":"European journal of cancer (Oxford, England : 1990)","url":"https://pubmed.ncbi.nlm.nih.gov/19695862","citation_count":8,"is_preprint":false},{"pmid":"34665359","id":"PMC_34665359","title":"Docking and ADMET studies for investigating the anticancer potency of Moscatilin on APC10/DOC1 and PKM2 against five clinical drugs.","date":"2021","source":"Journal, genetic engineering & biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/34665359","citation_count":7,"is_preprint":false},{"pmid":"31562243","id":"PMC_31562243","title":"The pseudosubstrate inhibitor Acm1 inhibits the anaphase-promoting complex/cyclosome by combining high-affinity activator binding with disruption of Doc1/Apc10 function.","date":"2019","source":"The Journal of biological 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Hungarica","url":"https://pubmed.ncbi.nlm.nih.gov/18297794","citation_count":4,"is_preprint":false},{"pmid":"20232159","id":"PMC_20232159","title":"Sequence characterization and promoter identification of porcine APC10 gene.","date":"2010","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/20232159","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":9680,"output_tokens":3017,"usd":0.037147,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10375,"output_tokens":3090,"usd":0.064562,"stage2_stop_reason":"end_turn"},"total_usd":0.101709,"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\": 2010,\n      \"finding\": \"Cryo-EM structure of APC/C(Cdh1) bound to a D-box peptide at ~10 Å resolution revealed that Cdh1 and Apc10 together form a co-receptor for the D-box degron motif. Cdh1 repositions toward Apc10 within the central cavity of the APC/C, and NMR spectroscopy demonstrated direct D-box–Apc10 interactions, establishing that Apc10 directly contributes to D-box recognition alongside the co-activator.\",\n      \"method\": \"Single-particle cryo-EM, difference mapping, NMR spectroscopy\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — cryo-EM structure plus NMR with direct binding validation; multiple orthogonal methods in one rigorous study\",\n      \"pmids\": [\"21107322\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"Crystal structure of human APC10/DOC1 at 1.6 Å resolution revealed a beta-sandwich jellyroll fold homologous to ligand-binding domains of galactose oxidase and coagulation factor Va. Biochemical experiments further demonstrated that the C-terminus of APC10 directly binds CDC27/APC3, a TPR-repeat-containing APC subunit.\",\n      \"method\": \"X-ray crystallography (1.6 Å); biochemical binding assay (C-terminus of APC10 binding to CDC27/APC3)\",\n      \"journal\": \"Nature structural biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure plus biochemical binding assay, single lab but two orthogonal methods\",\n      \"pmids\": [\"11524682\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Crystal structure of S. cerevisiae Doc1/Apc10 at 2.2 Å resolution showed a beta-sandwich homologous to the galactose-binding domain of galactose oxidase, the C2 domain of coagulation factor, and XRCC1. Residues invariant across Doc1/Apc10 sequences, including a temperature-sensitive mitotic arrest mutant, map to a beta-sheet region proposed to mediate biomolecular interactions and APC ubiquitination function.\",\n      \"method\": \"X-ray crystallography (2.2 Å); comparative structural analysis with mutagenesis data mapping\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure combined with functional mutant mapping, single lab\",\n      \"pmids\": [\"11884135\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"Doc1/Apc10 was shown to be a stoichiometric subunit of the yeast APC throughout the cell cycle. Mutation of Doc1/Apc10 inactivates APC ubiquitination activity without destabilizing the complex. The orthologous human APC10 protein is also a genuine APC subunit in vertebrates (human and frog), and its cellular levels and APC association are not cell-cycle-regulated, as determined by biochemical fractionation and mass spectrometric analysis.\",\n      \"method\": \"Immunoprecipitation, biochemical fractionation, mass spectrometry, in vivo ubiquitination assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP, mass spectrometry, functional ubiquitination assays; replicated in yeast and vertebrates\",\n      \"pmids\": [\"10318877\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"In fission yeast, apc10+ is essential for viability and required for ubiquitination and degradation of mitotic B-type cyclins. apc10 mutants show temperature-sensitive growth with defects in chromosome segregation and fail to arrest at G1 upon nitrogen starvation. A subpopulation of Apc10 co-immunoprecipitates with the APC, though it does not co-sediment with the 20S complex, suggesting a regulatory association.\",\n      \"method\": \"Genetic mutant analysis, co-immunoprecipitation, cell cycle analysis, ubiquitination assay\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic loss-of-function with defined mitotic phenotype, co-IP, ubiquitination assay in fission yeast\",\n      \"pmids\": [\"9736616\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"Human APC10/Doc1 binds APC core subunits throughout the cell cycle and localizes to centrosomes and mitotic spindles during mitosis, to kinetochores from prophase to anaphase, and to the midbody during telophase/cytokinesis, as determined by co-immunoprecipitation and immunofluorescence localization studies.\",\n      \"method\": \"Co-immunoprecipitation; immunofluorescence microscopy (subcellular localization)\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — co-IP and direct localization imaging, single lab, two orthogonal methods but no functional consequence of localization directly tested\",\n      \"pmids\": [\"10498862\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"ANAPC10 physically interacts with Smad3 (via the MH2 domain), and together with CDH1 forms a complex with HEF1 (NEDD9). Domain mapping showed distinct Smad3 MH2 subdomains bind APC10 and HEF1. Overexpression of APC10 and CDH1 regulated HEF1 protein levels, suggesting Smad3 recruits the APC/C to HEF1 for ubiquitination and proteasomal degradation via direct Smad3–APC10 interaction.\",\n      \"method\": \"Co-immunoprecipitation, domain mapping, overexpression/protein level assay\",\n      \"journal\": \"BMC cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-IP and domain mapping, single lab, multiple interaction partners tested but no in vitro reconstitution\",\n      \"pmids\": [\"15144564\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"The pseudosubstrate APC/C inhibitor Acm1 from budding yeast suppresses APC/C activity by combining high-affinity Cdh1 binding with a C-terminal D-box extension that specifically disrupts the normal interaction with Doc1/Apc10, thereby perturbing reaction processivity in ubiquitylation. Mutation of the conserved D-box converted Acm1 into an ABBA-motif-dependent APC/CCdh1 substrate, and biochemical analysis confirmed the extension's role in inhibiting processivity via Doc1/Apc10.\",\n      \"method\": \"In vivo mutant analysis, biochemical APC/C activity assays, site-directed mutagenesis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — biochemical assay with mutagenesis and in vivo validation, single lab\",\n      \"pmids\": [\"31562243\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"Disruption of the mouse Apc10/Doc1 gene underlies the oligosyndactylism (Os) radiation-induced mutation and two transgene-induced alleles (94-A and 94-K), all exhibiting a cell-autonomous block in metaphase-to-anaphase transition, establishing that Apc10/Doc1 is required for this cell cycle transition in vivo.\",\n      \"method\": \"Genetic mapping, molecular characterization of three mutant alleles, cell cycle phenotype analysis\",\n      \"journal\": \"Genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — three independent loss-of-function alleles with defined mitotic arrest phenotype, single study\",\n      \"pmids\": [\"11247669\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"In Drosophila, loss-of-function mutations in Apc10/Doc1 cause metaphase-like arrest, chromosome overcondensation, high mitotic index, and accumulation of cyclin B (an APC/C substrate) in larval neuroblasts, establishing that Apc10/Doc1 is essential for APC/C E3 ubiquitin ligase activity and cyclin B ubiquitination in vivo.\",\n      \"method\": \"Genetic loss-of-function (mutant alleles), immunostaining for cyclin B, mitotic index quantification\",\n      \"journal\": \"Acta biologica Hungarica\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with defined substrate accumulation phenotype, single lab\",\n      \"pmids\": [\"18297794\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"During interphase, APC10 interacts with NLRP3 and promotes NLRP3 inflammasome activation; during mitosis, APC10 dissociates from NLRP3 to repress inflammatory responses, establishing a cell-cycle-dependent switch role for APC10 in innate immune signaling.\",\n      \"method\": \"Co-immunoprecipitation, cell cycle stage-specific analysis, inflammasome activation assays\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-IP plus functional inflammasome assay, single lab, two orthogonal approaches\",\n      \"pmids\": [\"34407203\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"ANAPC10 protein is mainly expressed in the cytoplasm of spermatogonia and leptotene/pachytene spermatocytes in the developing mouse testis, as established by immunofluorescence and in situ hybridization.\",\n      \"method\": \"In situ hybridization, immunofluorescence microscopy\",\n      \"journal\": \"Biology of reproduction\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, localization only, no direct functional consequence demonstrated for this tissue-specific localization\",\n      \"pmids\": [\"22190705\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ANAPC10 (APC10/DOC1) is a core, stoichiometric subunit of the APC/C ubiquitin ligase whose jellyroll beta-sandwich DOC domain directly contacts the destruction (D-box) degron of substrates in concert with the co-activator Cdh1, stabilizes the active holoenzyme for processive ubiquitylation, binds the APC subunit CDC27/APC3 via its C-terminus, and is essential for APC/C-dependent cyclin B ubiquitination and the metaphase-to-anaphase transition; it additionally interacts with Smad3 to recruit the APC/C for HEF1 degradation and, during interphase, binds NLRP3 to promote inflammasome activation in a cell-cycle-regulated manner.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ANAPC10 (APC10/DOC1) is a core, stoichiometric subunit of the anaphase-promoting complex/cyclosome (APC/C) E3 ubiquitin ligase that is required for substrate degron recognition and the metaphase-to-anaphase transition [#3, #8]. Its structure is a beta-sandwich jellyroll DOC domain, and its C-terminus directly binds the TPR-repeat APC subunit CDC27/APC3 to anchor it within the complex [#1, #2]. Functionally, APC10 together with the co-activator Cdh1 forms a co-receptor for the destruction-box (D-box) degron: cryo-EM positions Cdh1 adjacent to Apc10 in the central APC/C cavity, and NMR demonstrates direct D-box–Apc10 contacts, explaining why APC10 is needed for processive substrate ubiquitylation rather than for complex assembly [#0, #7]. Loss of APC10 inactivates APC/C ubiquitination activity without destabilizing the complex, causing cell-autonomous metaphase arrest and accumulation of the substrate cyclin B across yeast, fly, and mouse systems [#3, #4, #9]. Beyond its constitutive APC/C role, APC10 interacts with Smad3 to recruit the APC/C–CDH1 ligase for degradation of HEF1/NEDD9 [#6], and during interphase binds NLRP3 to promote inflammasome activation, dissociating during mitosis—a cell-cycle-gated function in innate immune signaling [#10].\",\n  \"teleology\": [\n    {\n      \"year\": 1998,\n      \"claim\": \"Established that Apc10/Doc1 is essential for APC/C-mediated mitotic cyclin destruction, linking it directly to chromosome segregation control.\",\n      \"evidence\": \"Genetic loss-of-function, co-IP, and ubiquitination assays in fission yeast\",\n      \"pmids\": [\"9736616\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of D-box recognition not yet defined\", \"Only a subpopulation co-IPs with APC, leaving stoichiometry ambiguous at the time\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Resolved whether Apc10 is a regulatory factor or a true subunit by showing it is a stoichiometric, cell-cycle-invariant APC component whose mutation abolishes ubiquitination without disassembling the complex.\",\n      \"evidence\": \"Reciprocal co-IP, biochemical fractionation, mass spectrometry, and in vivo ubiquitination assays in yeast and vertebrates\",\n      \"pmids\": [\"10318877\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular basis of how a stable subunit contributes to catalysis unresolved\", \"No structural model\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Provided spatial context by mapping human APC10 to centrosomes, spindles, kinetochores, and the midbody through mitosis.\",\n      \"evidence\": \"Co-IP and immunofluorescence microscopy in human cells\",\n      \"pmids\": [\"10498862\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of each localization not tested\", \"Single lab\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Defined the APC10 fold as a beta-sandwich jellyroll resembling ligand-binding domains and showed its C-terminus binds CDC27/APC3, explaining how it is tethered within the APC/C.\",\n      \"evidence\": \"1.6 Å X-ray crystallography plus biochemical binding assay\",\n      \"pmids\": [\"11524682\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not identify the ligand engaged by the jellyroll surface\", \"D-box binding not yet demonstrated\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Demonstrated in mammals that Apc10/Doc1 disruption causes a cell-autonomous metaphase-to-anaphase block, confirming in vivo requirement in a vertebrate.\",\n      \"evidence\": \"Genetic mapping and characterization of three mouse mutant alleles with cell cycle phenotyping\",\n      \"pmids\": [\"11247669\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Substrate-level mechanism not directly assayed\", \"Single study\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Mapped functionally essential, invariant residues—including a temperature-sensitive arrest mutant—onto a beta-sheet surface, predicting the interaction face used for APC ubiquitination.\",\n      \"evidence\": \"2.2 Å crystallography of yeast Doc1 with mutagenesis mapping\",\n      \"pmids\": [\"11884135\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The predicted interaction partner of the beta-sheet face not yet identified structurally\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Extended APC10 function beyond the core cell cycle by showing Smad3 directly binds APC10 to recruit the APC/C–CDH1 ligase for HEF1/NEDD9 degradation.\",\n      \"evidence\": \"Co-IP, domain mapping, and overexpression protein-level assays\",\n      \"pmids\": [\"15144564\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No in vitro reconstitution of HEF1 ubiquitination\", \"Single lab\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Settled how APC10 contributes to specificity by showing it acts with Cdh1 as a bipartite D-box co-receptor, with direct Apc10–D-box contacts within the APC/C cavity.\",\n      \"evidence\": \"Single-particle cryo-EM with difference mapping and NMR of D-box–Apc10 binding\",\n      \"pmids\": [\"21107322\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Atomic-resolution contacts limited by ~10 Å EM\", \"Generality across diverse D-box substrates not exhaustively tested\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Reinforced the processivity role by showing the inhibitor Acm1 uses a D-box extension to specifically disrupt the Doc1/Apc10 interaction, confirming Apc10–degron engagement drives processive ubiquitylation.\",\n      \"evidence\": \"In vivo mutant analysis, biochemical APC/C activity assays, and site-directed mutagenesis in budding yeast\",\n      \"pmids\": [\"31562243\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Quantitative kinetics of processivity loss not fully resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identified a moonlighting role in which APC10 binds NLRP3 in interphase to promote inflammasome activation and dissociates in mitosis, coupling cell cycle phase to innate immune output.\",\n      \"evidence\": \"Co-IP and cell-cycle stage-specific inflammasome activation assays\",\n      \"pmids\": [\"34407203\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether this requires APC/C catalytic activity or is APC/C-independent unresolved\", \"Single lab, no reciprocal in vivo validation\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How APC10's non-canonical interactions (Smad3/HEF1, NLRP3) are mechanistically integrated with—or separated from—its constitutive APC/C role remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No reconstituted system distinguishing APC/C-dependent from APC/C-independent APC10 activities\", \"Physiological significance of testis-restricted expression uncharacterized\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 3, 4, 9]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [11]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [3, 4, 8, 9]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 3, 6]}\n    ],\n    \"complexes\": [\"APC/C (anaphase-promoting complex/cyclosome)\"],\n    \"partners\": [\"CDC27\", \"CDH1\", \"SMAD3\", \"NEDD9\", \"NLRP3\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}