{"gene":"FBXO5","run_date":"2026-06-09T23:54:43","timeline":{"discoveries":[{"year":2001,"finding":"EMI1 (FBXO5) binds the APC activator Cdc20 and inhibits APC/C ubiquitin ligase activity; it contains a zinc-binding region (ZBR) essential for APC inhibition; immunodepletion from cycling Xenopus extracts delays cyclin B accumulation and mitotic entry, while non-destructible EMI1 stabilizes APC substrates and causes mitotic block.","method":"Xenopus extract immunodepletion, gain-of-function rescue, Co-immunoprecipitation, domain mutagenesis (ZBR)","journal":"Cell","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal methods (immunodepletion, reconstitution in Xenopus extracts, domain mutagenesis, Co-IP); foundational paper, independently replicated in subsequent studies","pmids":["11389834"],"is_preprint":false},{"year":2002,"finding":"EMI1 is necessary and sufficient to inhibit APC/C(Cdc20) and maintain CSF (cytostatic factor) metaphase II arrest in Xenopus eggs; immunodepletion of EMI1 from CSF extracts causes premature cyclin B degradation and mitotic exit in the absence of calcium.","method":"Xenopus egg extract immunodepletion, add-back rescue experiments","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — reconstitution in Xenopus egg extracts, reciprocal add-back rescue, replicated context from founding lab","pmids":["11976684"],"is_preprint":false},{"year":2003,"finding":"EMI1 is phosphorylated by Cdc2 on a DSGxxS consensus degron, which is then recognized by the SCF(β-TrCP/Slimb) ubiquitin ligase, leading to EMI1 ubiquitylation and destruction in prophase; failure of β-TrCP-dependent EMI1 destruction stabilizes APC substrates and causes mitotic catastrophe including centrosome overduplication.","method":"Phosphorylation assays, SCF(β-TrCP) binding assays, loss-of-function in cells and Drosophila slimb mutants, ubiquitination assays","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro phosphorylation and ubiquitination assays combined with cell-based and genetic (Drosophila) validation; independently replicated by Hansen et al. 2004","pmids":["12791267"],"is_preprint":false},{"year":2004,"finding":"Polo-like kinase 1 (Plk1) is strictly required for EMI1 destruction: Plk1 phosphorylates EMI1 to generate the DSGxxS degron recognized by SCF(β-TrCP), stimulating β-TrCP binding and ubiquitination in vitro; cyclin B/Cdk1 enhances these effects. Plk1 binds EMI1 in mitosis and both proteins co-localize on mitotic spindle poles.","method":"In vitro kinase assays, ubiquitination assays, Co-immunoprecipitation, co-localization by immunofluorescence, Plk1 overexpression and dominant-negative approaches","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro reconstitution (kinase + ubiquitination assays) plus reciprocal Co-IP and co-localization; multiple orthogonal methods in single study, confirmatory of Margottin-Goguet 2003","pmids":["15469984"],"is_preprint":false},{"year":2006,"finding":"EMI1 acts as a pseudosubstrate inhibitor of APC/C(Cdh1): it stably binds both APC/C and its Cdh1 activator, competes with APC substrates at the D-box receptor site via its C-terminal D-box (providing binding affinity), and antagonizes E3 ligase activity via its ZBR independent of tight APC binding. Mutation of the ZBR converts EMI1 into a D-box-dependent APC substrate.","method":"In vitro binding assays, APC ubiquitination assays, mutagenesis of D-box and ZBR, competition assays with APC substrates","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with mutagenesis of multiple domains; mechanistic model confirmed by multiple orthogonal biochemical assays","pmids":["16921029"],"is_preprint":false},{"year":2006,"finding":"The Evi5 oncogene binds EMI1 adjacent to its DSGxxS degron and stabilizes EMI1 by blocking degron phosphorylation by Polo-like kinases and subsequent β-TrCP binding, thereby antagonizing SCF(β-TrCP)-dependent EMI1 ubiquitination; Evi5 depletion causes precocious EMI1 degradation, premature APC/C activation, cyclin destruction, and mitotic catastrophe.","method":"Co-immunoprecipitation (Evi5-EMI1 interaction), in vitro ubiquitination assays, siRNA knockdown with cell cycle phenotyping","journal":"Cell","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — reciprocal Co-IP, in vitro ubiquitination rescue assays, and KD phenotype with multiple cell cycle readouts","pmids":["16439210"],"is_preprint":false},{"year":2006,"finding":"Pin1 stabilizes EMI1 during G2 phase by binding EMI1 and preventing its association with SCF(β-TrCP) in an isomerization-dependent manner; Pin1-EMI1 interaction is detectable in vivo during G2, protecting EMI1 from degradation at this cell cycle phase.","method":"Co-immunoprecipitation in Xenopus XL2 cells, Pin1-EMI1 binding assays, isomerization-dependent competition with β-TrCP, loss-of-function cell cycle analysis","journal":"EMBO reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP in vivo during G2 plus functional rescue; single lab, two orthogonal methods","pmids":["17159919"],"is_preprint":false},{"year":2007,"finding":"EMI1 is essential for preventing rereplication: EMI1 depletion causes premature APC/C activation, destabilizing geminin and cyclin A (which redundantly prevent rereplication in mammalian cells); cyclin A's anti-rereplication role is mediated through its association with Cdk2 and Cdk1.","method":"siRNA knockdown, rescue experiments with non-degradable geminin/cyclin A, flow cytometry (rereplication readout), co-immunoprecipitation","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis (double depletion rescue), multiple KD/rescue with defined molecular mechanism; replicated across multiple studies","pmids":["17234884"],"is_preprint":false},{"year":2007,"finding":"EMI1 degradation is NOT required to activate APC/C at mitotic entry; instead, EMI1's essential function is to inhibit APC/C in interphase to stabilize mitotic cyclins and geminin to promote mitotic entry and prevent rereplication.","method":"Live-cell imaging of EMI1 degradation timing vs. APC/C activation, siRNA knockdown, cell cycle analysis","journal":"The Journal of cell biology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — single lab, live-cell imaging with functional readouts; contradicts some prior claims; single method set","pmids":["17485488"],"is_preprint":false},{"year":2013,"finding":"EMI1's 143-residue C-terminal domain inhibits APC/C(CDH1) through multiple mechanisms simultaneously: the intrinsically disordered D-box, linker, and tail elements together with the structured ZBR bind distinct regions of APC/C(CDH1) to synergistically block the substrate-binding site and inhibit ubiquitin-chain elongation.","method":"NMR, cryo-electron microscopy, enzymology (APC/C ubiquitination assays), domain deletion/mutation analysis","journal":"Nature structural & molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — hybrid structural approach (NMR + EM) combined with enzyme assays and domain mapping; multiple orthogonal methods in single rigorous study","pmids":["23708605"],"is_preprint":false},{"year":2013,"finding":"EMI1 inhibits APC ubiquitylation at two distinct steps: (1) substrate binding and (2) ubiquitin chain elongation; the ZBR allows multiple monoubiquitylation of substrates but preferentially suppresses ubiquitin chain elongation by UBCH10; the C-terminal tail of EMI1 antagonizes chain elongation by Ube2S through competitive inhibition of its binding to the APC cullin subunit via electrostatic interaction.","method":"In vitro APC ubiquitination kinetic assays, domain mutagenesis, competition binding assays with Ube2S","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with kinetic analysis and mutagenesis; single lab but multiple orthogonal biochemical approaches","pmids":["23708001"],"is_preprint":false},{"year":2018,"finding":"Cell-cycle commitment at the G1/S transition is mediated by an EMI1-APC/C(CDH1) dual-negative feedback switch: EMI1 transitions from being a substrate of APC/C(CDH1) to being its inhibitor, triggered by increased CDK2 activity and EMI1 mRNA expression. In vitro reconstitution showed this dual-negative feedback is bistable and irreversible.","method":"Human cell models with live-cell reporters, in vitro reconstitution of EMI1-APC/C switch, mathematical modeling, siRNA knockdown, cell synchronization","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro reconstitution plus live-cell functional analysis, multiple orthogonal methods, high-impact venue","pmids":["29875408"],"is_preprint":false},{"year":2018,"finding":"EMI1's F-box domain is required for assembly of a canonical SCF ubiquitin ligase complex that constitutively targets RAD51 for proteasomal degradation; in response to genotoxic stress, CHK1-mediated phosphorylation of RAD51 counteracts EMI1-dependent degradation by enhancing RAD51's affinity for BRCA2. EMI1 depletion restores RAD51 accumulation and homologous recombination repair in BRCA1-deficient cells.","method":"Genetic screen for PARPi sensitivity, F-box domain mutagenesis, Co-immunoprecipitation (EMI1-RAD51 SCF complex), ubiquitination assays, siRNA knockdown, orthotopic mouse model","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — genetic screen, domain mutagenesis (ΔF-box), biochemical ubiquitination assays, in vivo mouse model; multiple orthogonal methods","pmids":["30554948"],"is_preprint":false},{"year":2004,"finding":"p90Rsk2 associates with and phosphorylates EMI1 upstream of the Cdc20 binding region, stabilizing the EMI1-Cdc20 interaction and contributing to metaphase arrest in mouse oocytes; the Mos-MAPK pathway establishes CSF activity through stabilization of an APC-inhibitory complex composed of EMI1 and Cdc20.","method":"Co-immunoprecipitation (p90Rsk2-EMI1), in vitro phosphorylation assays, RNA interference, transfection into two-cell embryos","journal":"The EMBO journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and in vitro kinase assay plus functional RNAi rescue; single lab, two orthogonal methods","pmids":["15526037"],"is_preprint":false},{"year":2011,"finding":"Bcr-Abl increases EMI1 stability through Src kinase-mediated tyrosine phosphorylation of EMI1 at Tyr142; mutation Y142F abolishes phosphorylation by recombinant Src kinase and reduces EMI1 stability. Stable EMI1 prevents APC/Cdh1-mediated ubiquitination of Skp2, thereby maintaining Skp2 protein levels in CML cells.","method":"Kinase inhibitor treatment, site-directed mutagenesis (Y142F), in vitro phosphorylation by recombinant Src, Co-immunoprecipitation, ubiquitination assays, half-life analysis","journal":"Journal of cellular physiology","confidence":"Medium","confidence_rationale":"Tier 1–2 / Moderate — in vitro phosphorylation with mutagenesis plus cellular ubiquitination assays; single lab, two orthogonal methods","pmids":["20717963"],"is_preprint":false},{"year":2024,"finding":"FBXO5 directly binds RNF183 and promotes its ubiquitin-dependent proteasomal degradation through its F-box domain; this FBXO5/RNF183 axis regulates ER stress-induced apoptosis. F-box domain deletion (ΔF-box mutant) abolishes the anti-apoptotic effect, establishing requirement for SCF complex assembly.","method":"Co-immunoprecipitation (FBXO5-RNF183), ubiquitination assays, F-box deletion mutagenesis, siRNA knockdown, in vivo tumor xenograft","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assays, F-box mutagenesis, in vivo validation; single lab but multiple orthogonal methods","pmids":["38212299"],"is_preprint":false},{"year":2021,"finding":"PUMA associates with EMI1 and RAD51 in the cytoplasm, facilitating EMI1-mediated cytoplasmic RAD51 ubiquitination and degradation, thereby inhibiting RAD51 nuclear translocation and homologous recombination DNA repair in embryonic and hematopoietic progenitor cells.","method":"Co-immunoprecipitation (PUMA-EMI1-RAD51), ubiquitination assays, subcellular fractionation, nuclear translocation assays, HR repair assays","journal":"Signal transduction and targeted therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assays, and functional HR readout; single lab, multiple orthogonal methods","pmids":["33785736"],"is_preprint":false},{"year":2009,"finding":"EMI1 down-regulation by p21(WAF1) after DNA damage contributes to APC/C activation, resulting in degradation of cyclins A2 and B1 and maintenance of G2 arrest; siRNA-mediated EMI1 depletion prevents irradiated p21-deficient cells from entering mitosis, and this effect is counteracted by APC/C inactivation.","method":"siRNA knockdown, genetic epistasis (p21+/+ vs p21-/- cells), flow cytometry (G2 arrest), Western blot for cyclin levels","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with double KD rescue; single lab, multiple cell line contexts","pmids":["19211842"],"is_preprint":false},{"year":2024,"finding":"Cryo-EM structures of human APC/C(CDH1):EMI1 ternary complex at 2.9 Å resolution reveal the N-terminus of CDH1 (CDH1α1) at the APC/C interface; a zinc-binding module in APC2 confers structural stability; intrinsically disordered regions of multiple APC/C subunits involved in assembly and regulation are resolved, confirming multimodal EMI1 binding.","method":"Cryo-electron microscopy, AlphaFold-assisted model building, experimental zinc ion detection","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1 / Moderate — high-resolution cryo-EM structure (2.9 Å) with experimental validation of zinc; single study but structurally definitive","pmids":["39567505"],"is_preprint":false},{"year":2022,"finding":"Using comprehensive enzyme assays, the context-dependent roles of multiple EMI1 C-terminal motifs were determined: EMI1 motifs inhibit both APC/C substrate recruitment and APC/C-associated E2s (UBE2C and UBE2S); additionally, an isolated C-terminal peptide fragment of EMI1 activates APC/C-dependent substrate priming by UBE2C.","method":"In vitro ubiquitination assays with truncation and point mutants of EMI1, APC/C reconstitution","journal":"Protein science","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — in vitro reconstitution with domain analysis; single lab, comprehensive but single publication","pmids":["35634770"],"is_preprint":false},{"year":2025,"finding":"FBXO5 mediates K48-linked ubiquitination and proteasomal degradation of SARS-CoV-2 NSP7; NSP7 ubiquitination is co-regulated by β-TrCP1 and the kinase TAF1. A small molecule that disrupts the β-TrCP1-FBXO5 interaction stabilizes FBXO5 and enhances NSP7 degradation.","method":"esiRNA screen, ubiquitination assays (K48-linkage specific), Co-immunoprecipitation (β-TrCP1-FBXO5), small molecule stabilizer screen, SARS-CoV-2 infection assays","journal":"Advanced science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, linkage-specific ubiquitination assays, functional antiviral readout; single lab, multiple orthogonal methods","pmids":["40577599"],"is_preprint":false},{"year":2025,"finding":"FBXO5 promotes K48-linked polyubiquitination and proteasomal degradation of DOK6; DOK6 depletion enhances mechanical rigidity in GBM tumor cells and initiates survival signaling that confers temozolomide resistance.","method":"CRISPR/Cas9 sgRNA library screen, Co-immunoprecipitation, ubiquitination assays (K48 linkage), FBXO5 knockdown/overexpression, in vitro and in vivo tumor models","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CRISPR screen plus Co-IP and ubiquitination assays with in vivo validation; single lab, multiple orthogonal methods","pmids":["41045986"],"is_preprint":false},{"year":2025,"finding":"FBXO5 facilitates TP53 ubiquitination-mediated degradation; Co-IP confirmed direct interaction between FBXO5 and TP53; CHX chase and ubiquitination assays established that FBXO5 reduces TP53 protein stability via the proteasome pathway, promoting osteogenic differentiation of human stem cells from the apical papilla.","method":"Co-immunoprecipitation, cycloheximide chase assay, ubiquitination assay, siRNA knockdown, osteogenic differentiation assays","journal":"International dental journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, CHX chase, and ubiquitination assays; single lab, three orthogonal biochemical methods","pmids":["41240879"],"is_preprint":false},{"year":2005,"finding":"EMI1 is required for the meiosis I to meiosis II (MI-MII) transition in Xenopus oocytes: acute antibody-mediated neutralization of EMI1 immediately after GVBD causes rapid loss of Cdc2 activity with simultaneous cyclin B loss and MAPK inactivation, causing chromosome decondensation and DNA replication instead of MII progression; these defects are rescued by non-destructible cyclin B, Cdc20 depletion, or methyl-ubiquitin.","method":"Neutralizing antibody injection in Xenopus oocytes, rescue with Delta90 cyclin B, Cdc20 depletion, methyl-ubiquitin, cell cycle phase analysis","journal":"Cell cycle","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — antibody neutralization plus multiple genetic epistasis rescues in Xenopus; single lab, multiple rescue conditions","pmids":["15701974"],"is_preprint":false},{"year":2009,"finding":"EMI1 depletion in human cells or zebrafish embryos causes chromosomal rereplication, leading to rereplicated unsegregated chromosomes and polyploidy; EMI1-depleted mammalian cells rely on topoisomerase IIα-dependent mitotic decatenation to progress through metaphase.","method":"Morpholino knockdown in zebrafish, siRNA in human cells, flow cytometry, metaphase chromosome preparation, BrdU incorporation, topoisomerase IIα inhibition","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — morpholino KD in vivo plus siRNA KD with defined genomic readouts; single lab, cross-species validation","pmids":["19704007"],"is_preprint":false}],"current_model":"FBXO5/EMI1 is an F-box protein that functions as a potent, multimodal pseudosubstrate inhibitor of the APC/C ubiquitin ligase: it binds APC/C and its co-activators (Cdc20, Cdh1) via a C-terminal D-box, blocks substrate access, and suppresses ubiquitin chain elongation via its zinc-binding region (ZBR) and C-terminal tail; it is regulated by SCF(β-TrCP)-mediated destruction triggered by sequential Plk1/Cdk1 phosphorylation of its DSGxxS degron, stabilized by Evi5 and Pin1 during S/G2, and—through its F-box domain—assembles SCF complexes that target substrates including RAD51, RNF183, DOK6, and TP53 for ubiquitin-dependent degradation, establishing roles in coordinating DNA replication with mitosis, preventing rereplication, maintaining genomic stability, regulating DNA repair, and modulating responses to viral infection."},"narrative":{"mechanistic_narrative":"FBXO5/EMI1 is a cell-cycle regulator that couples DNA replication to mitosis by acting as a potent, multimodal pseudosubstrate inhibitor of the anaphase-promoting complex/cyclosome (APC/C) [PMID:11389834, PMID:16921029]. It stably engages APC/C and its co-activators Cdc20 and Cdh1, using a C-terminal D-box to compete with substrates at the D-box receptor while its zinc-binding region (ZBR) and C-terminal tail simultaneously suppress ubiquitin-chain elongation by the E2 enzymes UBCH10/UBE2C and Ube2S [PMID:16921029, PMID:23708605, PMID:23708001]; cryo-EM of the APC/C(CDH1):EMI1 ternary complex confirms this multimodal binding mode [PMID:39567505]. Through this inhibition EMI1 stabilizes mitotic cyclins and geminin during interphase to promote mitotic entry and prevent re-replication, and its loss triggers premature APC/C activation, geminin/cyclin A destabilization, rereplication, and polyploidy [PMID:17234884, PMID:17485488, PMID:19704007]. At the G1/S transition EMI1 switches from an APC/C(CDH1) substrate to its inhibitor, forming a bistable dual-negative feedback loop driven by rising CDK2 activity that enforces irreversible cell-cycle commitment [PMID:29875408]. EMI1 levels are tightly controlled by phosphorylation: sequential Plk1 and Cdk1/Cdc2 phosphorylation of a DSGxxS degron generates the recognition site for SCF(β-TrCP/Slimb)-mediated ubiquitylation and destruction, while Evi5 and Pin1 protect the degron during S/G2 [PMID:12791267, PMID:15469984, PMID:16439210, PMID:17159919]. Independently of its APC/C-inhibitory role, EMI1's own F-box domain nucleates a canonical SCF ubiquitin ligase that targets substrates for proteasomal degradation, including RAD51—where SCF-EMI1 restrains homologous recombination and modulates PARP-inhibitor and BRCA-deficient repair responses [PMID:30554948, PMID:33785736]—as well as RNF183 in ER-stress apoptosis, DOK6, TP53, and the SARS-CoV-2 protein NSP7 [PMID:38212299, PMID:40577599, PMID:41045986, PMID:41240879]. EMI1 also enforces metaphase II/cytostatic-factor arrest and the meiosis I–II transition in oocytes by stabilizing the cyclin B/Cdc20 axis [PMID:11976684, PMID:15701974].","teleology":[{"year":2001,"claim":"Established that EMI1 is an APC/C inhibitor controlling mitotic timing, answering how cyclin B is allowed to accumulate before mitosis.","evidence":"Xenopus extract immunodepletion, add-back rescue, Co-IP, and ZBR domain mutagenesis","pmids":["11389834"],"confidence":"High","gaps":["Molecular mechanism of APC/C inhibition (substrate competition vs. catalytic block) not yet resolved","Role across full cell cycle in somatic cells untested"]},{"year":2002,"claim":"Showed EMI1 is necessary and sufficient to maintain cytostatic-factor metaphase II arrest, defining its role in vertebrate egg cell-cycle arrest.","evidence":"Xenopus egg extract immunodepletion with reciprocal add-back rescue","pmids":["11976684"],"confidence":"High","gaps":["Upstream signals stabilizing EMI1 during CSF arrest unaddressed","Relationship to Mos-MAPK pathway not yet defined"]},{"year":2003,"claim":"Defined the destruction mechanism: Cdc2 phosphorylation of a DSGxxS degron recruits SCF(β-TrCP) to destroy EMI1 in prophase, explaining how APC/C inhibition is relieved.","evidence":"Phosphorylation and ubiquitination assays, β-TrCP binding, Drosophila slimb mutants","pmids":["12791267"],"confidence":"High","gaps":["Whether Cdc2 is the sole/priming kinase unresolved","Spatial regulation of degron phosphorylation unknown"]},{"year":2004,"claim":"Identified Plk1 as the strictly required degron kinase and established its co-localization with EMI1 at spindle poles, refining the phosphorylation hierarchy for EMI1 destruction.","evidence":"In vitro kinase and ubiquitination assays, Co-IP, immunofluorescence co-localization, dominant-negative Plk1","pmids":["15469984"],"confidence":"High","gaps":["Order of Plk1 vs. Cdk1 phosphorylation not fully separated","In vivo requirement in mammalian cells limited"]},{"year":2004,"claim":"Linked the Mos-MAPK pathway to EMI1 by showing p90Rsk2 phosphorylates EMI1 to stabilize the EMI1-Cdc20 inhibitory complex during oocyte metaphase arrest.","evidence":"Co-IP, in vitro kinase assay, RNAi, two-cell embryo transfection in mouse oocytes","pmids":["15526037"],"confidence":"Medium","gaps":["Single-lab finding with two orthogonal methods","Phosphosite mapping incomplete","Generalizability beyond oocytes untested"]},{"year":2005,"claim":"Demonstrated EMI1 is required for the meiosis I to meiosis II transition, extending its arrest function to meiotic progression.","evidence":"Neutralizing antibody injection in Xenopus oocytes with cyclin B, Cdc20-depletion, and methyl-ubiquitin rescues","pmids":["15701974"],"confidence":"Medium","gaps":["Antibody neutralization may have off-target effects","Mechanism distinguishing MI vs. MII control unclear"]},{"year":2006,"claim":"Defined EMI1 as a pseudosubstrate inhibitor of APC/C(Cdh1), separating D-box-mediated binding affinity from ZBR-mediated catalytic inhibition.","evidence":"In vitro binding and APC ubiquitination assays with D-box and ZBR mutagenesis and substrate competition","pmids":["16921029"],"confidence":"High","gaps":["Structural basis of multimodal binding not yet resolved","Contribution of C-terminal tail not yet dissected"]},{"year":2006,"claim":"Identified Evi5 as a stabilizer of EMI1 that blocks degron phosphorylation, revealing positive regulation of EMI1 abundance during the cell cycle.","evidence":"Reciprocal Co-IP, in vitro ubiquitination rescue, siRNA with cell-cycle phenotyping","pmids":["16439210"],"confidence":"High","gaps":["Precise cell-cycle window of Evi5 action partially defined","Coordination with Pin1 stabilization unresolved"]},{"year":2006,"claim":"Showed Pin1 stabilizes EMI1 in G2 via isomerization-dependent blocking of β-TrCP binding, adding a phase-specific layer of EMI1 protection.","evidence":"Co-IP in Xenopus XL2 cells, isomerization-dependent β-TrCP competition, loss-of-function cell-cycle analysis","pmids":["17159919"],"confidence":"Medium","gaps":["Single-lab finding","Pin1 target proline residue not definitively mapped"]},{"year":2007,"claim":"Established that EMI1 prevents rereplication by stabilizing geminin and cyclin A through APC/C inhibition, defining its genome-protective function.","evidence":"siRNA with non-degradable geminin/cyclin A rescue, flow cytometry, Co-IP","pmids":["17234884"],"confidence":"High","gaps":["Relative contribution of geminin vs. cyclin A context-dependent","Mechanism of cyclin A anti-rereplication action partly inferred"]},{"year":2007,"claim":"Clarified that EMI1 destruction is not required to activate APC/C at mitotic entry, repositioning EMI1's essential role to interphase APC/C inhibition.","evidence":"Live-cell imaging of degradation timing vs. APC/C activation, siRNA, cell-cycle analysis","pmids":["17485488"],"confidence":"Medium","gaps":["Single method set; contradicts some prior models","Does not exclude partial roles for EMI1 turnover at mitosis"]},{"year":2009,"claim":"Connected EMI1 to the DNA damage response by showing p21-mediated EMI1 downregulation activates APC/C to enforce G2 arrest.","evidence":"siRNA, p21+/+ vs p21-/- epistasis, flow cytometry, cyclin Western blots","pmids":["19211842"],"confidence":"Medium","gaps":["Mechanism of p21-driven EMI1 loss not defined","Single-lab finding"]},{"year":2009,"claim":"Showed EMI1 depletion causes rereplication and polyploidy in human cells and zebrafish and creates dependence on topoisomerase IIα decatenation, demonstrating in vivo genome-stability roles.","evidence":"Zebrafish morpholino and human siRNA knockdown, flow cytometry, metaphase chromosome preparation, topoisomerase inhibition","pmids":["19704007"],"confidence":"Medium","gaps":["Single-lab finding","Molecular basis of topoisomerase dependence not fully resolved"]},{"year":2011,"claim":"Linked oncogenic signaling to EMI1 stability by showing Bcr-Abl/Src tyrosine phosphorylation at Tyr142 stabilizes EMI1 and sustains Skp2 in CML cells.","evidence":"Kinase inhibitors, Y142F mutagenesis, in vitro Src phosphorylation, Co-IP, ubiquitination and half-life assays","pmids":["20717963"],"confidence":"Medium","gaps":["Single-lab finding","In vivo relevance of Tyr142 in normal cells untested"]},{"year":2013,"claim":"Resolved the structural and kinetic basis of multimodal APC/C inhibition, showing the C-terminal domain blocks substrate binding while ZBR and tail suppress chain elongation by distinct E2s.","evidence":"NMR, cryo-EM, APC/C ubiquitination kinetics, domain mutagenesis (two complementary studies)","pmids":["23708605","23708001"],"confidence":"High","gaps":["Atomic-resolution view of the full ternary complex not yet achieved","Regulation of these motifs by phosphorylation unaddressed"]},{"year":2018,"claim":"Defined the EMI1–APC/C(CDH1) dual-negative feedback switch as the bistable basis of irreversible G1/S commitment, integrating EMI1 regulation into cell-cycle decision-making.","evidence":"Live-cell reporters, in vitro reconstitution of the switch, mathematical modeling, siRNA","pmids":["29875408"],"confidence":"High","gaps":["Quantitative thresholds in vivo across cell types not mapped","Coupling to mitogen signaling partly inferred"]},{"year":2018,"claim":"Established a distinct SCF-EMI1 ligase function: the F-box domain assembles an SCF that degrades RAD51 to restrain homologous recombination, with implications for PARP-inhibitor sensitivity in BRCA-deficient cells.","evidence":"PARPi-sensitivity genetic screen, ΔF-box mutagenesis, Co-IP, ubiquitination assays, siRNA, orthotopic mouse model","pmids":["30554948"],"confidence":"High","gaps":["Whether SCF-EMI1 and APC/C-inhibitory functions are temporally separated unclear","Full substrate repertoire of SCF-EMI1 unknown"]},{"year":2021,"claim":"Showed PUMA scaffolds cytoplasmic EMI1-mediated RAD51 ubiquitination to block RAD51 nuclear import and HR in progenitor cells, adding a subcellular dimension to EMI1's repair regulation.","evidence":"Co-IP, ubiquitination assays, subcellular fractionation, nuclear translocation and HR repair assays","pmids":["33785736"],"confidence":"Medium","gaps":["Single-lab finding","Direct vs. scaffolded EMI1-RAD51 contact not separated from PUMA role"]},{"year":2022,"claim":"Mapped context-dependent activities of individual EMI1 C-terminal motifs, finding inhibition of substrate recruitment and E2 activity but paradoxical activation of UBE2C priming by an isolated peptide.","evidence":"In vitro ubiquitination assays with EMI1 truncations and point mutants, APC/C reconstitution","pmids":["35634770"],"confidence":"Medium","gaps":["Physiological relevance of the activating peptide fragment unknown","Single comprehensive in vitro study"]},{"year":2024,"claim":"Provided a 2.9 Å cryo-EM structure of the APC/C(CDH1):EMI1 ternary complex, confirming multimodal EMI1 binding and resolving regulatory disordered regions and an APC2 zinc module.","evidence":"Cryo-EM with AlphaFold-assisted modeling and experimental zinc detection","pmids":["39567505"],"confidence":"High","gaps":["Dynamics of the inhibitory-to-substrate transition not captured","Structures with phosphorylated EMI1 absent"]},{"year":2024,"claim":"Extended SCF-EMI1 substrate repertoire to RNF183, linking the FBXO5/RNF183 axis to control of ER stress-induced apoptosis.","evidence":"Co-IP, ubiquitination assays, ΔF-box mutagenesis, siRNA, tumor xenograft","pmids":["38212299"],"confidence":"Medium","gaps":["Single-lab finding","How ER stress regulates this axis undefined"]},{"year":2025,"claim":"Identified additional SCF-EMI1 substrates DOK6 and TP53 and an antiviral role degrading SARS-CoV-2 NSP7, broadening FBXO5 ligase function into glioma drug resistance, stem-cell differentiation, and infection.","evidence":"CRISPR/esiRNA screens, Co-IP, K48-linkage ubiquitination assays, CHX chase, in vivo tumor models, SARS-CoV-2 infection assays","pmids":["41045986","41240879","40577599"],"confidence":"Medium","gaps":["Each substrate reported by a single lab","Degron/recognition motifs for these substrates not defined","Relationship to cell-cycle SCF-EMI1 function unclear"]},{"year":null,"claim":"How EMI1's APC/C-inhibitory and SCF-ligase functions are temporally, spatially, and structurally partitioned within a single protein remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of the F-box/SCF-EMI1 complex with substrates","Degron recognition logic for the expanding SCF-EMI1 substrate set undefined","Switch governing inhibitor vs. ligase mode unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[12,15,20,21,22]},{"term_id":"GO:0098772","term_label":"molecular function regulator 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During mitotic cell cycle plays a role as both substrate and inhibitor of APC-FZR1 complex (PubMed:16921029, PubMed:17234884, PubMed:17485488, PubMed:17875940, PubMed:23708001, PubMed:23708605, PubMed:29875408). During G1 phase, plays a role as substrate of APC-FZR1 complex E3 ligase (PubMed:29875408). Then switches as an inhibitor of APC-FZR1 complex during S and G2 leading to cell-cycle commitment (PubMed:29875408). As APC inhibitor, prevents the degradation of APC substrates at multiple levels: by interacting with APC and blocking access of APC substrates to the D-box coreceptor, formed by FZR1 and ANAPC10; by suppressing ubiquitin ligation and chain elongation by APC by preventing the UBE2C and UBE2S activities (PubMed:16921029, PubMed:23708001, PubMed:23708605). Plays a role in genome integrity preservation by coordinating DNA replication with mitosis through APC inhibition in interphase to stabilize CCNA2 and GMNN in order to promote mitosis and prevent rereplication and DNA damage-induced cellular senescence (PubMed:17234884, PubMed:17485488, PubMed:17875940). During oocyte maturation, plays a role in meiosis through inactivation of APC-FZR1 complex. Inhibits APC through RPS6KA2 interaction that increases FBXO5 affiniy for CDC20 leading to the metaphase arrest of the second meiotic division before fertilization (By similarity). Controls entry into the first meiotic division through inactivation of APC-FZR1 complex (By similarity). 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FBXO5","url":"https://www.omim.org/entry/606013"},{"mim_id":"602942","title":"ECOTROPIC VIRAL INTEGRATION SITE 5; EVI5","url":"https://www.omim.org/entry/602942"},{"mim_id":"142957","title":"HOMEOBOX A10; HOXA10","url":"https://www.omim.org/entry/142957"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"}],"tissue_specificity":"Group enriched","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"bone marrow","ntpm":41.6},{"tissue":"lymphoid 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block.\",\n      \"method\": \"Xenopus extract immunodepletion, gain-of-function rescue, Co-immunoprecipitation, domain mutagenesis (ZBR)\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal methods (immunodepletion, reconstitution in Xenopus extracts, domain mutagenesis, Co-IP); foundational paper, independently replicated in subsequent studies\",\n      \"pmids\": [\"11389834\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"EMI1 is necessary and sufficient to inhibit APC/C(Cdc20) and maintain CSF (cytostatic factor) metaphase II arrest in Xenopus eggs; immunodepletion of EMI1 from CSF extracts causes premature cyclin B degradation and mitotic exit in the absence of calcium.\",\n      \"method\": \"Xenopus egg extract immunodepletion, add-back rescue experiments\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — reconstitution in Xenopus egg extracts, reciprocal add-back rescue, replicated context from founding lab\",\n      \"pmids\": [\"11976684\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"EMI1 is phosphorylated by Cdc2 on a DSGxxS consensus degron, which is then recognized by the SCF(β-TrCP/Slimb) ubiquitin ligase, leading to EMI1 ubiquitylation and destruction in prophase; failure of β-TrCP-dependent EMI1 destruction stabilizes APC substrates and causes mitotic catastrophe including centrosome overduplication.\",\n      \"method\": \"Phosphorylation assays, SCF(β-TrCP) binding assays, loss-of-function in cells and Drosophila slimb mutants, ubiquitination assays\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro phosphorylation and ubiquitination assays combined with cell-based and genetic (Drosophila) validation; independently replicated by Hansen et al. 2004\",\n      \"pmids\": [\"12791267\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Polo-like kinase 1 (Plk1) is strictly required for EMI1 destruction: Plk1 phosphorylates EMI1 to generate the DSGxxS degron recognized by SCF(β-TrCP), stimulating β-TrCP binding and ubiquitination in vitro; cyclin B/Cdk1 enhances these effects. Plk1 binds EMI1 in mitosis and both proteins co-localize on mitotic spindle poles.\",\n      \"method\": \"In vitro kinase assays, ubiquitination assays, Co-immunoprecipitation, co-localization by immunofluorescence, Plk1 overexpression and dominant-negative approaches\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro reconstitution (kinase + ubiquitination assays) plus reciprocal Co-IP and co-localization; multiple orthogonal methods in single study, confirmatory of Margottin-Goguet 2003\",\n      \"pmids\": [\"15469984\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"EMI1 acts as a pseudosubstrate inhibitor of APC/C(Cdh1): it stably binds both APC/C and its Cdh1 activator, competes with APC substrates at the D-box receptor site via its C-terminal D-box (providing binding affinity), and antagonizes E3 ligase activity via its ZBR independent of tight APC binding. Mutation of the ZBR converts EMI1 into a D-box-dependent APC substrate.\",\n      \"method\": \"In vitro binding assays, APC ubiquitination assays, mutagenesis of D-box and ZBR, competition assays with APC substrates\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with mutagenesis of multiple domains; mechanistic model confirmed by multiple orthogonal biochemical assays\",\n      \"pmids\": [\"16921029\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"The Evi5 oncogene binds EMI1 adjacent to its DSGxxS degron and stabilizes EMI1 by blocking degron phosphorylation by Polo-like kinases and subsequent β-TrCP binding, thereby antagonizing SCF(β-TrCP)-dependent EMI1 ubiquitination; Evi5 depletion causes precocious EMI1 degradation, premature APC/C activation, cyclin destruction, and mitotic catastrophe.\",\n      \"method\": \"Co-immunoprecipitation (Evi5-EMI1 interaction), in vitro ubiquitination assays, siRNA knockdown with cell cycle phenotyping\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — reciprocal Co-IP, in vitro ubiquitination rescue assays, and KD phenotype with multiple cell cycle readouts\",\n      \"pmids\": [\"16439210\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Pin1 stabilizes EMI1 during G2 phase by binding EMI1 and preventing its association with SCF(β-TrCP) in an isomerization-dependent manner; Pin1-EMI1 interaction is detectable in vivo during G2, protecting EMI1 from degradation at this cell cycle phase.\",\n      \"method\": \"Co-immunoprecipitation in Xenopus XL2 cells, Pin1-EMI1 binding assays, isomerization-dependent competition with β-TrCP, loss-of-function cell cycle analysis\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP in vivo during G2 plus functional rescue; single lab, two orthogonal methods\",\n      \"pmids\": [\"17159919\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"EMI1 is essential for preventing rereplication: EMI1 depletion causes premature APC/C activation, destabilizing geminin and cyclin A (which redundantly prevent rereplication in mammalian cells); cyclin A's anti-rereplication role is mediated through its association with Cdk2 and Cdk1.\",\n      \"method\": \"siRNA knockdown, rescue experiments with non-degradable geminin/cyclin A, flow cytometry (rereplication readout), co-immunoprecipitation\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis (double depletion rescue), multiple KD/rescue with defined molecular mechanism; replicated across multiple studies\",\n      \"pmids\": [\"17234884\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"EMI1 degradation is NOT required to activate APC/C at mitotic entry; instead, EMI1's essential function is to inhibit APC/C in interphase to stabilize mitotic cyclins and geminin to promote mitotic entry and prevent rereplication.\",\n      \"method\": \"Live-cell imaging of EMI1 degradation timing vs. APC/C activation, siRNA knockdown, cell cycle analysis\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — single lab, live-cell imaging with functional readouts; contradicts some prior claims; single method set\",\n      \"pmids\": [\"17485488\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"EMI1's 143-residue C-terminal domain inhibits APC/C(CDH1) through multiple mechanisms simultaneously: the intrinsically disordered D-box, linker, and tail elements together with the structured ZBR bind distinct regions of APC/C(CDH1) to synergistically block the substrate-binding site and inhibit ubiquitin-chain elongation.\",\n      \"method\": \"NMR, cryo-electron microscopy, enzymology (APC/C ubiquitination assays), domain deletion/mutation analysis\",\n      \"journal\": \"Nature structural & molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — hybrid structural approach (NMR + EM) combined with enzyme assays and domain mapping; multiple orthogonal methods in single rigorous study\",\n      \"pmids\": [\"23708605\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"EMI1 inhibits APC ubiquitylation at two distinct steps: (1) substrate binding and (2) ubiquitin chain elongation; the ZBR allows multiple monoubiquitylation of substrates but preferentially suppresses ubiquitin chain elongation by UBCH10; the C-terminal tail of EMI1 antagonizes chain elongation by Ube2S through competitive inhibition of its binding to the APC cullin subunit via electrostatic interaction.\",\n      \"method\": \"In vitro APC ubiquitination kinetic assays, domain mutagenesis, competition binding assays with Ube2S\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with kinetic analysis and mutagenesis; single lab but multiple orthogonal biochemical approaches\",\n      \"pmids\": [\"23708001\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Cell-cycle commitment at the G1/S transition is mediated by an EMI1-APC/C(CDH1) dual-negative feedback switch: EMI1 transitions from being a substrate of APC/C(CDH1) to being its inhibitor, triggered by increased CDK2 activity and EMI1 mRNA expression. In vitro reconstitution showed this dual-negative feedback is bistable and irreversible.\",\n      \"method\": \"Human cell models with live-cell reporters, in vitro reconstitution of EMI1-APC/C switch, mathematical modeling, siRNA knockdown, cell synchronization\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro reconstitution plus live-cell functional analysis, multiple orthogonal methods, high-impact venue\",\n      \"pmids\": [\"29875408\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"EMI1's F-box domain is required for assembly of a canonical SCF ubiquitin ligase complex that constitutively targets RAD51 for proteasomal degradation; in response to genotoxic stress, CHK1-mediated phosphorylation of RAD51 counteracts EMI1-dependent degradation by enhancing RAD51's affinity for BRCA2. EMI1 depletion restores RAD51 accumulation and homologous recombination repair in BRCA1-deficient cells.\",\n      \"method\": \"Genetic screen for PARPi sensitivity, F-box domain mutagenesis, Co-immunoprecipitation (EMI1-RAD51 SCF complex), ubiquitination assays, siRNA knockdown, orthotopic mouse model\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — genetic screen, domain mutagenesis (ΔF-box), biochemical ubiquitination assays, in vivo mouse model; multiple orthogonal methods\",\n      \"pmids\": [\"30554948\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"p90Rsk2 associates with and phosphorylates EMI1 upstream of the Cdc20 binding region, stabilizing the EMI1-Cdc20 interaction and contributing to metaphase arrest in mouse oocytes; the Mos-MAPK pathway establishes CSF activity through stabilization of an APC-inhibitory complex composed of EMI1 and Cdc20.\",\n      \"method\": \"Co-immunoprecipitation (p90Rsk2-EMI1), in vitro phosphorylation assays, RNA interference, transfection into two-cell embryos\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and in vitro kinase assay plus functional RNAi rescue; single lab, two orthogonal methods\",\n      \"pmids\": [\"15526037\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Bcr-Abl increases EMI1 stability through Src kinase-mediated tyrosine phosphorylation of EMI1 at Tyr142; mutation Y142F abolishes phosphorylation by recombinant Src kinase and reduces EMI1 stability. Stable EMI1 prevents APC/Cdh1-mediated ubiquitination of Skp2, thereby maintaining Skp2 protein levels in CML cells.\",\n      \"method\": \"Kinase inhibitor treatment, site-directed mutagenesis (Y142F), in vitro phosphorylation by recombinant Src, Co-immunoprecipitation, ubiquitination assays, half-life analysis\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro phosphorylation with mutagenesis plus cellular ubiquitination assays; single lab, two orthogonal methods\",\n      \"pmids\": [\"20717963\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"FBXO5 directly binds RNF183 and promotes its ubiquitin-dependent proteasomal degradation through its F-box domain; this FBXO5/RNF183 axis regulates ER stress-induced apoptosis. F-box domain deletion (ΔF-box mutant) abolishes the anti-apoptotic effect, establishing requirement for SCF complex assembly.\",\n      \"method\": \"Co-immunoprecipitation (FBXO5-RNF183), ubiquitination assays, F-box deletion mutagenesis, siRNA knockdown, in vivo tumor xenograft\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assays, F-box mutagenesis, in vivo validation; single lab but multiple orthogonal methods\",\n      \"pmids\": [\"38212299\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"PUMA associates with EMI1 and RAD51 in the cytoplasm, facilitating EMI1-mediated cytoplasmic RAD51 ubiquitination and degradation, thereby inhibiting RAD51 nuclear translocation and homologous recombination DNA repair in embryonic and hematopoietic progenitor cells.\",\n      \"method\": \"Co-immunoprecipitation (PUMA-EMI1-RAD51), ubiquitination assays, subcellular fractionation, nuclear translocation assays, HR repair assays\",\n      \"journal\": \"Signal transduction and targeted therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assays, and functional HR readout; single lab, multiple orthogonal methods\",\n      \"pmids\": [\"33785736\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"EMI1 down-regulation by p21(WAF1) after DNA damage contributes to APC/C activation, resulting in degradation of cyclins A2 and B1 and maintenance of G2 arrest; siRNA-mediated EMI1 depletion prevents irradiated p21-deficient cells from entering mitosis, and this effect is counteracted by APC/C inactivation.\",\n      \"method\": \"siRNA knockdown, genetic epistasis (p21+/+ vs p21-/- cells), flow cytometry (G2 arrest), Western blot for cyclin levels\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with double KD rescue; single lab, multiple cell line contexts\",\n      \"pmids\": [\"19211842\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Cryo-EM structures of human APC/C(CDH1):EMI1 ternary complex at 2.9 Å resolution reveal the N-terminus of CDH1 (CDH1α1) at the APC/C interface; a zinc-binding module in APC2 confers structural stability; intrinsically disordered regions of multiple APC/C subunits involved in assembly and regulation are resolved, confirming multimodal EMI1 binding.\",\n      \"method\": \"Cryo-electron microscopy, AlphaFold-assisted model building, experimental zinc ion detection\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — high-resolution cryo-EM structure (2.9 Å) with experimental validation of zinc; single study but structurally definitive\",\n      \"pmids\": [\"39567505\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Using comprehensive enzyme assays, the context-dependent roles of multiple EMI1 C-terminal motifs were determined: EMI1 motifs inhibit both APC/C substrate recruitment and APC/C-associated E2s (UBE2C and UBE2S); additionally, an isolated C-terminal peptide fragment of EMI1 activates APC/C-dependent substrate priming by UBE2C.\",\n      \"method\": \"In vitro ubiquitination assays with truncation and point mutants of EMI1, APC/C reconstitution\",\n      \"journal\": \"Protein science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — in vitro reconstitution with domain analysis; single lab, comprehensive but single publication\",\n      \"pmids\": [\"35634770\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"FBXO5 mediates K48-linked ubiquitination and proteasomal degradation of SARS-CoV-2 NSP7; NSP7 ubiquitination is co-regulated by β-TrCP1 and the kinase TAF1. A small molecule that disrupts the β-TrCP1-FBXO5 interaction stabilizes FBXO5 and enhances NSP7 degradation.\",\n      \"method\": \"esiRNA screen, ubiquitination assays (K48-linkage specific), Co-immunoprecipitation (β-TrCP1-FBXO5), small molecule stabilizer screen, SARS-CoV-2 infection assays\",\n      \"journal\": \"Advanced science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, linkage-specific ubiquitination assays, functional antiviral readout; single lab, multiple orthogonal methods\",\n      \"pmids\": [\"40577599\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"FBXO5 promotes K48-linked polyubiquitination and proteasomal degradation of DOK6; DOK6 depletion enhances mechanical rigidity in GBM tumor cells and initiates survival signaling that confers temozolomide resistance.\",\n      \"method\": \"CRISPR/Cas9 sgRNA library screen, Co-immunoprecipitation, ubiquitination assays (K48 linkage), FBXO5 knockdown/overexpression, in vitro and in vivo tumor models\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CRISPR screen plus Co-IP and ubiquitination assays with in vivo validation; single lab, multiple orthogonal methods\",\n      \"pmids\": [\"41045986\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"FBXO5 facilitates TP53 ubiquitination-mediated degradation; Co-IP confirmed direct interaction between FBXO5 and TP53; CHX chase and ubiquitination assays established that FBXO5 reduces TP53 protein stability via the proteasome pathway, promoting osteogenic differentiation of human stem cells from the apical papilla.\",\n      \"method\": \"Co-immunoprecipitation, cycloheximide chase assay, ubiquitination assay, siRNA knockdown, osteogenic differentiation assays\",\n      \"journal\": \"International dental journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, CHX chase, and ubiquitination assays; single lab, three orthogonal biochemical methods\",\n      \"pmids\": [\"41240879\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"EMI1 is required for the meiosis I to meiosis II (MI-MII) transition in Xenopus oocytes: acute antibody-mediated neutralization of EMI1 immediately after GVBD causes rapid loss of Cdc2 activity with simultaneous cyclin B loss and MAPK inactivation, causing chromosome decondensation and DNA replication instead of MII progression; these defects are rescued by non-destructible cyclin B, Cdc20 depletion, or methyl-ubiquitin.\",\n      \"method\": \"Neutralizing antibody injection in Xenopus oocytes, rescue with Delta90 cyclin B, Cdc20 depletion, methyl-ubiquitin, cell cycle phase analysis\",\n      \"journal\": \"Cell cycle\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — antibody neutralization plus multiple genetic epistasis rescues in Xenopus; single lab, multiple rescue conditions\",\n      \"pmids\": [\"15701974\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"EMI1 depletion in human cells or zebrafish embryos causes chromosomal rereplication, leading to rereplicated unsegregated chromosomes and polyploidy; EMI1-depleted mammalian cells rely on topoisomerase IIα-dependent mitotic decatenation to progress through metaphase.\",\n      \"method\": \"Morpholino knockdown in zebrafish, siRNA in human cells, flow cytometry, metaphase chromosome preparation, BrdU incorporation, topoisomerase IIα inhibition\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — morpholino KD in vivo plus siRNA KD with defined genomic readouts; single lab, cross-species validation\",\n      \"pmids\": [\"19704007\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"FBXO5/EMI1 is an F-box protein that functions as a potent, multimodal pseudosubstrate inhibitor of the APC/C ubiquitin ligase: it binds APC/C and its co-activators (Cdc20, Cdh1) via a C-terminal D-box, blocks substrate access, and suppresses ubiquitin chain elongation via its zinc-binding region (ZBR) and C-terminal tail; it is regulated by SCF(β-TrCP)-mediated destruction triggered by sequential Plk1/Cdk1 phosphorylation of its DSGxxS degron, stabilized by Evi5 and Pin1 during S/G2, and—through its F-box domain—assembles SCF complexes that target substrates including RAD51, RNF183, DOK6, and TP53 for ubiquitin-dependent degradation, establishing roles in coordinating DNA replication with mitosis, preventing rereplication, maintaining genomic stability, regulating DNA repair, and modulating responses to viral infection.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"FBXO5/EMI1 is a cell-cycle regulator that couples DNA replication to mitosis by acting as a potent, multimodal pseudosubstrate inhibitor of the anaphase-promoting complex/cyclosome (APC/C) [#0, #4]. It stably engages APC/C and its co-activators Cdc20 and Cdh1, using a C-terminal D-box to compete with substrates at the D-box receptor while its zinc-binding region (ZBR) and C-terminal tail simultaneously suppress ubiquitin-chain elongation by the E2 enzymes UBCH10/UBE2C and Ube2S [#4, #9, #10]; cryo-EM of the APC/C(CDH1):EMI1 ternary complex confirms this multimodal binding mode [#18]. Through this inhibition EMI1 stabilizes mitotic cyclins and geminin during interphase to promote mitotic entry and prevent re-replication, and its loss triggers premature APC/C activation, geminin/cyclin A destabilization, rereplication, and polyploidy [#7, #8, #24]. At the G1/S transition EMI1 switches from an APC/C(CDH1) substrate to its inhibitor, forming a bistable dual-negative feedback loop driven by rising CDK2 activity that enforces irreversible cell-cycle commitment [#11]. EMI1 levels are tightly controlled by phosphorylation: sequential Plk1 and Cdk1/Cdc2 phosphorylation of a DSGxxS degron generates the recognition site for SCF(\\u03b2-TrCP/Slimb)-mediated ubiquitylation and destruction, while Evi5 and Pin1 protect the degron during S/G2 [#2, #3, #5, #6]. Independently of its APC/C-inhibitory role, EMI1's own F-box domain nucleates a canonical SCF ubiquitin ligase that targets substrates for proteasomal degradation, including RAD51\\u2014where SCF-EMI1 restrains homologous recombination and modulates PARP-inhibitor and BRCA-deficient repair responses [#12, #16]\\u2014as well as RNF183 in ER-stress apoptosis, DOK6, TP53, and the SARS-CoV-2 protein NSP7 [#15, #20, #21, #22]. EMI1 also enforces metaphase II/cytostatic-factor arrest and the meiosis I\\u2013II transition in oocytes by stabilizing the cyclin B/Cdc20 axis [#1, #23].\",\n  \"teleology\": [\n    {\n      \"year\": 2001,\n      \"claim\": \"Established that EMI1 is an APC/C inhibitor controlling mitotic timing, answering how cyclin B is allowed to accumulate before mitosis.\",\n      \"evidence\": \"Xenopus extract immunodepletion, add-back rescue, Co-IP, and ZBR domain mutagenesis\",\n      \"pmids\": [\"11389834\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism of APC/C inhibition (substrate competition vs. catalytic block) not yet resolved\", \"Role across full cell cycle in somatic cells untested\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Showed EMI1 is necessary and sufficient to maintain cytostatic-factor metaphase II arrest, defining its role in vertebrate egg cell-cycle arrest.\",\n      \"evidence\": \"Xenopus egg extract immunodepletion with reciprocal add-back rescue\",\n      \"pmids\": [\"11976684\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Upstream signals stabilizing EMI1 during CSF arrest unaddressed\", \"Relationship to Mos-MAPK pathway not yet defined\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Defined the destruction mechanism: Cdc2 phosphorylation of a DSGxxS degron recruits SCF(\\u03b2-TrCP) to destroy EMI1 in prophase, explaining how APC/C inhibition is relieved.\",\n      \"evidence\": \"Phosphorylation and ubiquitination assays, \\u03b2-TrCP binding, Drosophila slimb mutants\",\n      \"pmids\": [\"12791267\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether Cdc2 is the sole/priming kinase unresolved\", \"Spatial regulation of degron phosphorylation unknown\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Identified Plk1 as the strictly required degron kinase and established its co-localization with EMI1 at spindle poles, refining the phosphorylation hierarchy for EMI1 destruction.\",\n      \"evidence\": \"In vitro kinase and ubiquitination assays, Co-IP, immunofluorescence co-localization, dominant-negative Plk1\",\n      \"pmids\": [\"15469984\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Order of Plk1 vs. Cdk1 phosphorylation not fully separated\", \"In vivo requirement in mammalian cells limited\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Linked the Mos-MAPK pathway to EMI1 by showing p90Rsk2 phosphorylates EMI1 to stabilize the EMI1-Cdc20 inhibitory complex during oocyte metaphase arrest.\",\n      \"evidence\": \"Co-IP, in vitro kinase assay, RNAi, two-cell embryo transfection in mouse oocytes\",\n      \"pmids\": [\"15526037\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab finding with two orthogonal methods\", \"Phosphosite mapping incomplete\", \"Generalizability beyond oocytes untested\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Demonstrated EMI1 is required for the meiosis I to meiosis II transition, extending its arrest function to meiotic progression.\",\n      \"evidence\": \"Neutralizing antibody injection in Xenopus oocytes with cyclin B, Cdc20-depletion, and methyl-ubiquitin rescues\",\n      \"pmids\": [\"15701974\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Antibody neutralization may have off-target effects\", \"Mechanism distinguishing MI vs. MII control unclear\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Defined EMI1 as a pseudosubstrate inhibitor of APC/C(Cdh1), separating D-box-mediated binding affinity from ZBR-mediated catalytic inhibition.\",\n      \"evidence\": \"In vitro binding and APC ubiquitination assays with D-box and ZBR mutagenesis and substrate competition\",\n      \"pmids\": [\"16921029\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of multimodal binding not yet resolved\", \"Contribution of C-terminal tail not yet dissected\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Identified Evi5 as a stabilizer of EMI1 that blocks degron phosphorylation, revealing positive regulation of EMI1 abundance during the cell cycle.\",\n      \"evidence\": \"Reciprocal Co-IP, in vitro ubiquitination rescue, siRNA with cell-cycle phenotyping\",\n      \"pmids\": [\"16439210\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Precise cell-cycle window of Evi5 action partially defined\", \"Coordination with Pin1 stabilization unresolved\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Showed Pin1 stabilizes EMI1 in G2 via isomerization-dependent blocking of \\u03b2-TrCP binding, adding a phase-specific layer of EMI1 protection.\",\n      \"evidence\": \"Co-IP in Xenopus XL2 cells, isomerization-dependent \\u03b2-TrCP competition, loss-of-function cell-cycle analysis\",\n      \"pmids\": [\"17159919\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab finding\", \"Pin1 target proline residue not definitively mapped\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Established that EMI1 prevents rereplication by stabilizing geminin and cyclin A through APC/C inhibition, defining its genome-protective function.\",\n      \"evidence\": \"siRNA with non-degradable geminin/cyclin A rescue, flow cytometry, Co-IP\",\n      \"pmids\": [\"17234884\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative contribution of geminin vs. cyclin A context-dependent\", \"Mechanism of cyclin A anti-rereplication action partly inferred\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Clarified that EMI1 destruction is not required to activate APC/C at mitotic entry, repositioning EMI1's essential role to interphase APC/C inhibition.\",\n      \"evidence\": \"Live-cell imaging of degradation timing vs. APC/C activation, siRNA, cell-cycle analysis\",\n      \"pmids\": [\"17485488\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single method set; contradicts some prior models\", \"Does not exclude partial roles for EMI1 turnover at mitosis\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Connected EMI1 to the DNA damage response by showing p21-mediated EMI1 downregulation activates APC/C to enforce G2 arrest.\",\n      \"evidence\": \"siRNA, p21+/+ vs p21-/- epistasis, flow cytometry, cyclin Western blots\",\n      \"pmids\": [\"19211842\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of p21-driven EMI1 loss not defined\", \"Single-lab finding\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Showed EMI1 depletion causes rereplication and polyploidy in human cells and zebrafish and creates dependence on topoisomerase II\\u03b1 decatenation, demonstrating in vivo genome-stability roles.\",\n      \"evidence\": \"Zebrafish morpholino and human siRNA knockdown, flow cytometry, metaphase chromosome preparation, topoisomerase inhibition\",\n      \"pmids\": [\"19704007\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab finding\", \"Molecular basis of topoisomerase dependence not fully resolved\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Linked oncogenic signaling to EMI1 stability by showing Bcr-Abl/Src tyrosine phosphorylation at Tyr142 stabilizes EMI1 and sustains Skp2 in CML cells.\",\n      \"evidence\": \"Kinase inhibitors, Y142F mutagenesis, in vitro Src phosphorylation, Co-IP, ubiquitination and half-life assays\",\n      \"pmids\": [\"20717963\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab finding\", \"In vivo relevance of Tyr142 in normal cells untested\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Resolved the structural and kinetic basis of multimodal APC/C inhibition, showing the C-terminal domain blocks substrate binding while ZBR and tail suppress chain elongation by distinct E2s.\",\n      \"evidence\": \"NMR, cryo-EM, APC/C ubiquitination kinetics, domain mutagenesis (two complementary studies)\",\n      \"pmids\": [\"23708605\", \"23708001\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Atomic-resolution view of the full ternary complex not yet achieved\", \"Regulation of these motifs by phosphorylation unaddressed\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Defined the EMI1\\u2013APC/C(CDH1) dual-negative feedback switch as the bistable basis of irreversible G1/S commitment, integrating EMI1 regulation into cell-cycle decision-making.\",\n      \"evidence\": \"Live-cell reporters, in vitro reconstitution of the switch, mathematical modeling, siRNA\",\n      \"pmids\": [\"29875408\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Quantitative thresholds in vivo across cell types not mapped\", \"Coupling to mitogen signaling partly inferred\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Established a distinct SCF-EMI1 ligase function: the F-box domain assembles an SCF that degrades RAD51 to restrain homologous recombination, with implications for PARP-inhibitor sensitivity in BRCA-deficient cells.\",\n      \"evidence\": \"PARPi-sensitivity genetic screen, \\u0394F-box mutagenesis, Co-IP, ubiquitination assays, siRNA, orthotopic mouse model\",\n      \"pmids\": [\"30554948\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether SCF-EMI1 and APC/C-inhibitory functions are temporally separated unclear\", \"Full substrate repertoire of SCF-EMI1 unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Showed PUMA scaffolds cytoplasmic EMI1-mediated RAD51 ubiquitination to block RAD51 nuclear import and HR in progenitor cells, adding a subcellular dimension to EMI1's repair regulation.\",\n      \"evidence\": \"Co-IP, ubiquitination assays, subcellular fractionation, nuclear translocation and HR repair assays\",\n      \"pmids\": [\"33785736\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab finding\", \"Direct vs. scaffolded EMI1-RAD51 contact not separated from PUMA role\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Mapped context-dependent activities of individual EMI1 C-terminal motifs, finding inhibition of substrate recruitment and E2 activity but paradoxical activation of UBE2C priming by an isolated peptide.\",\n      \"evidence\": \"In vitro ubiquitination assays with EMI1 truncations and point mutants, APC/C reconstitution\",\n      \"pmids\": [\"35634770\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Physiological relevance of the activating peptide fragment unknown\", \"Single comprehensive in vitro study\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Provided a 2.9 \\u00c5 cryo-EM structure of the APC/C(CDH1):EMI1 ternary complex, confirming multimodal EMI1 binding and resolving regulatory disordered regions and an APC2 zinc module.\",\n      \"evidence\": \"Cryo-EM with AlphaFold-assisted modeling and experimental zinc detection\",\n      \"pmids\": [\"39567505\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Dynamics of the inhibitory-to-substrate transition not captured\", \"Structures with phosphorylated EMI1 absent\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Extended SCF-EMI1 substrate repertoire to RNF183, linking the FBXO5/RNF183 axis to control of ER stress-induced apoptosis.\",\n      \"evidence\": \"Co-IP, ubiquitination assays, \\u0394F-box mutagenesis, siRNA, tumor xenograft\",\n      \"pmids\": [\"38212299\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab finding\", \"How ER stress regulates this axis undefined\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identified additional SCF-EMI1 substrates DOK6 and TP53 and an antiviral role degrading SARS-CoV-2 NSP7, broadening FBXO5 ligase function into glioma drug resistance, stem-cell differentiation, and infection.\",\n      \"evidence\": \"CRISPR/esiRNA screens, Co-IP, K48-linkage ubiquitination assays, CHX chase, in vivo tumor models, SARS-CoV-2 infection assays\",\n      \"pmids\": [\"41045986\", \"41240879\", \"40577599\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Each substrate reported by a single lab\", \"Degron/recognition motifs for these substrates not defined\", \"Relationship to cell-cycle SCF-EMI1 function unclear\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How EMI1's APC/C-inhibitory and SCF-ligase functions are temporally, spatially, and structurally partitioned within a single protein remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of the F-box/SCF-EMI1 complex with substrates\", \"Degron recognition logic for the expanding SCF-EMI1 substrate set undefined\", \"Switch governing inhibitor vs. ligase mode unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [12, 15, 20, 21, 22]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 4, 9, 10]},\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [12, 15, 21]},\n      {\"term_id\": \"GO:0140313\", \"supporting_discovery_ids\": [4, 9]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [16]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [7]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 2, 7, 11]},\n      {\"term_id\": \"R-HSA-69306\", \"supporting_discovery_ids\": [7, 24]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [12, 16]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 4, 12]},\n      {\"term_id\": \"R-HSA-1474165\", \"supporting_discovery_ids\": [1, 23]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [15]}\n    ],\n    \"complexes\": [\"APC/C (as bound inhibitor)\", \"SCF (Skp1-Cul1-F-box)\"],\n    \"partners\": [\"CDC20\", \"CDH1/FZR1\", \"BTRC (\\u03b2-TrCP)\", \"PLK1\", \"EVI5\", \"PIN1\", \"RAD51\", \"TP53\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}