{"gene":"KCTD10","run_date":"2026-06-10T02:59:49","timeline":{"discoveries":[{"year":2005,"finding":"KCTD10 (rat) interacts with the small subunit of DNA polymerase delta and PCNA, identifying it as a member of the PDIP1 gene family involved in DNA replication/repair.","method":"Protein interaction assays (by analogy with PDIP1 family characterization; cloning and expression analysis)","journal":"Biochimica et biophysica acta","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — initial cloning paper with interaction data, replicated in subsequent studies","pmids":["15982757"],"is_preprint":false},{"year":2009,"finding":"KCTD10 interacts with PCNA (confirmed by yeast two-hybrid and co-immunoprecipitation), and its knockdown inhibits cell proliferation in A549 cells; nuclear localization was observed.","method":"Yeast two-hybrid, co-immunoprecipitation, siRNA knockdown with cell proliferation assay, immunofluorescence","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — reciprocal Co-IP plus functional KD assay, single lab","pmids":["19125419"],"is_preprint":false},{"year":2009,"finding":"SP1 binding to the proximal KCTD10 promoter stimulates KCTD10 transcription, while AP-2alpha binding to the same region represses it, as established by deletion mutagenesis, site-directed mutation, ChIP, and reporter assays.","method":"Deletion mutagenesis, site-directed mutagenesis, chromatin immunoprecipitation (ChIP), luciferase reporter assay, qRT-PCR, western blot","journal":"The FEBS journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (ChIP, mutagenesis, reporter assay) in single lab with rigorous controls","pmids":["19154347"],"is_preprint":false},{"year":2012,"finding":"TNFAIP1 interacts with KCTD10 and promotes ubiquitin-mediated proteasomal degradation of KCTD10 protein; both KCTD10 and TNFAIP1 inhibit NF-κB and AP-1 transcriptional activity.","method":"Yeast two-hybrid, GST pull-down, co-immunoprecipitation, colocalization, protein degradation assay, ubiquitination assay with MG132, luciferase reporter assay","journal":"Molecular biology reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (Y2H, GST pull-down, Co-IP, ubiquitination assay), single lab","pmids":["22810651"],"is_preprint":false},{"year":2014,"finding":"In zebrafish, Kctd10 directly binds Tbx5a (T-box transcription factor) and represses its transcriptional activity; loss of kctd10 causes atrioventricular canal defects rescued by tbx5a or has2 knockdown.","method":"Zebrafish mutant genetic screen, morpholino knockdown epistasis, protein binding assay (Kctd10-Tbx5 direct binding), in situ hybridization for cardiac markers","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis in vivo plus direct protein binding assay, multiple orthogonal approaches","pmids":["24430697"],"is_preprint":false},{"year":2014,"finding":"KCTD10 interacts with Cullin3 and Notch1 simultaneously, mediating Notch1 proteolytic degradation; KCTD10 knockout mice die ~E10.5 with severe angiogenesis and heart defects, and Notch signaling members are upregulated in KCTD10-deficient embryos and KCTD10-silenced HUVECs.","method":"KCTD10 knockout mouse generation, co-immunoprecipitation (endogenous), qRT-PCR, western blotting, VEGF stimulation assays","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — endogenous Co-IP plus in vivo KO phenotype, single lab","pmids":["25401743"],"is_preprint":false},{"year":2014,"finding":"Zebrafish kctd10 knockdown causes pericardial edema, loss of heart formation, and loss of intersomitic vessels; the heart defect is linked to RhoA and PCNA pathways.","method":"Morpholino knockdown in zebrafish, in situ hybridization for cardiac markers (amhc, vmhc, cmlc2), Flk-1 staining for vasculature","journal":"Acta biochimica et biophysica Sinica","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KD with defined cellular/developmental phenotype and marker analysis, single lab","pmids":["24705121"],"is_preprint":false},{"year":2018,"finding":"The Cullin-3-Rbx1-KCTD10 E3 ligase complex catalyzes K63 polyubiquitination of RhoB at lysine 162 and 181, targeting RhoB to lysosomes and thereby regulating endothelial barrier integrity via control of RhoB-mediated endothelial cell contraction.","method":"Co-immunoprecipitation, ubiquitination assays (K63-specific), site-directed mutagenesis of RhoB lysines, lysosomal pathway inhibition, siRNA knockdown with endothelial barrier assay (TEER/permeability)","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro ubiquitination assay with mutagenesis, Co-IP, functional barrier assay, multiple orthogonal methods","pmids":["29358211"],"is_preprint":false},{"year":2019,"finding":"The CUL3/KCTD10 E3 ligase ubiquitinates RhoB for degradation, which relieves RhoB-mediated inhibition of Rac1 at the plasma membrane, enabling EGF-induced Rac1 activation, dorsal membrane ruffle formation, and cell proliferation in HER2-positive breast cancer cells.","method":"siRNA depletion of CUL3 or KCTD10, Rac1 activity assay, cell proliferation assay, membrane ruffle quantification, co-immunoprecipitation","journal":"Cancer science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional epistasis (CUL3/KCTD10 → RhoB → Rac1) with multiple assays, single lab","pmids":["30515933"],"is_preprint":false},{"year":2019,"finding":"The CUL3/KCTD10 complex forms a complex with EIF3D and catalyzes K27 polyubiquitination of EIF3D at lysine 153 and 275 residues in hepatocellular carcinoma HepG2 cells.","method":"Mass spectrometry (complex identification), co-immunoprecipitation, ubiquitination assay with K27-specific linkage, site-directed mutagenesis of EIF3D lysines, KCTD10 depletion","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — MS identification plus mutagenesis-confirmed ubiquitination sites, single lab","pmids":["31280863"],"is_preprint":false},{"year":2022,"finding":"KCTD10 acts as an upstream regulator of Notch signaling in brown adipose tissue; KCTD10 overexpression suppresses UCP1 expression and thermogenesis, while BAT-specific KCTD10 knockdown increases thermogenesis and cold tolerance; inhibiting Notch signaling restores KCTD10-overexpression-suppressed thermogenesis, placing KCTD10 upstream of Notch.","method":"BAT-specific knockdown mouse model, KCTD10 overexpression in BAT, UCP1 western blot, Notch inhibitor rescue experiment, metabolic phenotyping","journal":"The Journal of endocrinology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis in vivo (Notch inhibitor rescue) plus KD/OE with defined phenotype, single lab","pmids":["34854382"],"is_preprint":false},{"year":2024,"finding":"KCTD10 is the substrate-recognizing subunit of CRL3 E3 ligase that ubiquitinates SLC7A11 (cystine transporter); USP18 is the counteracting deubiquitylase. Upon cystine deprivation, KCTD10 protein decreases and USP18 increases, causing SLC7A11 accumulation. KCTD10 destabilizes SLC7A11 to reduce cystine uptake and promote ferroptosis.","method":"neddylation inhibitor (MLN4924) treatment, KCTD10 and USP18 siRNA/overexpression, ubiquitination assays, cystine uptake assay, ferroptosis assays, co-immunoprecipitation, in vivo tumor growth assay","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (Co-IP, ubiquitination assay, functional cystine/ferroptosis assays, in vivo), rigorous mechanistic dissection","pmids":["38959043"],"is_preprint":false},{"year":2024,"finding":"KCTD10 mediates ubiquitination-dependent degradation of KCTD13 protein in the developing cortex; KCTD10 deficiency causes KCTD13 accumulation, abnormal neuronal progenitor proliferation/differentiation, reduced deep-layer neurons, and motor deficits in brain-specific Kctd10 KO mice.","method":"Brain-specific Kctd10 knockout mice, co-immunoprecipitation (KCTD10-KCTD13 interaction screen), ubiquitination assay, cortical layer marker analysis, behavioral testing","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — in vivo KO with defined phenotype, Co-IP interaction, ubiquitination assay, multiple orthogonal methods in single rigorous study","pmids":["38489388"],"is_preprint":false},{"year":2024,"finding":"KCTD10 undergoes liquid-liquid phase separation (LLPS) mediated by its intrinsically disordered region (IDR); the schizophrenia-associated p.C124W mutation disrupts LLPS, impairing RHOB degradation and causing excessive RHOB accumulation in postsynaptic density, leading to synaptic abnormalities and schizophrenia-like behaviors in heterozygous mice.","method":"Heterozygous KCTD10 C124W knockin mice, LLPS assay, IDR deletion mutant, ubiquitination/degradation assay for RHOB, postsynaptic density fractionation, behavioral assays","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — in vivo knockin mice, LLPS mechanistic assays, domain mutant analysis, multiple orthogonal methods in single rigorous study","pmids":["39565307"],"is_preprint":false},{"year":2025,"finding":"The CUL3-KCTD10 E3 ligase acts as a bivalent sensor of co-directional transcription-replication conflicts (TRCs): KCTD10 interacts with both the replisome and transcription machinery simultaneously; upon detecting co-directional TRCs, KCTD10 complexes assemble higher-order assemblies recruiting CUL3 to ubiquitinate and remove the RNA polymerase elongation factor TCEA2, allowing replisome bypass. KCTD10 loss causes TCEA2 retention, TRC accumulation, and increased DNA damage.","method":"Co-immunoprecipitation (KCTD10 with replisome and transcription machinery), ubiquitination assay for TCEA2, TRC detection assay, DNA damage markers, KCTD10 KO cells","journal":"Nature","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (Co-IP with both complexes, ubiquitination assay, TRC assay, KO phenotype), peer-reviewed in Nature","pmids":["41062692"],"is_preprint":false},{"year":2025,"finding":"The BTB domain of KCTD10 interacts with Armadillo repeat domains 1-9 of β-catenin and facilitates K48-linked ubiquitin-dependent degradation of β-catenin, leading to downregulation of PD-L1; endothelial-specific Kctd10 knockout promotes lung cancer metastasis and tumor angiogenesis via β-catenin signaling.","method":"IP-mass spectrometry, co-immunoprecipitation, ubiquitination assay (K48-specific), domain mapping, Kctd10 endothelial-specific KO mice (Kctd10flox/flox CDH5-CreERT2), in vivo tumor assays","journal":"Frontiers in immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — IP-MS plus domain-mapping Co-IP, K48-specific ubiquitination assay, conditional KO in vivo, multiple orthogonal methods","pmids":["40873559"],"is_preprint":false},{"year":2025,"finding":"KCTD10 promotes ubiquitination and proteasomal degradation of IGF2BP1; KCTD10 overexpression reduces IGF2BP1-mediated m6A stabilization of IL-11 mRNA, thereby restraining immune evasion in lung adenocarcinoma.","method":"Ubiquitination assay, IGF2BP1 half-life detection, RNA pull-down (IGF2BP1-IL-11 mRNA), ELISA, western blot, co-immunoprecipitation, allograft experiment","journal":"British journal of pharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ubiquitination assay plus functional downstream readout (m6A/IL-11 axis), single lab with multiple methods","pmids":["42107984"],"is_preprint":false}],"current_model":"KCTD10 functions primarily as the substrate-recognition adaptor of the CUL3-RING E3 ubiquitin ligase complex, targeting diverse substrates (RhoB via K63-ubiquitination for lysosomal degradation, SLC7A11 for proteasomal degradation, EIF3D via K27-ubiquitination, KCTD13 for degradation, β-catenin via K48-ubiquitination, IGF2BP1, and the RNA Pol II elongation factor TCEA2) to regulate endothelial barrier function, ferroptosis, cancer cell proliferation, brain development, and genome stability at transcription-replication conflicts; it also undergoes liquid-liquid phase separation via its intrinsically disordered region to facilitate RhoB degradation at synapses, and can directly bind and repress transcription factors (Tbx5a) to control cardiac development, while negatively regulating Notch signaling in multiple developmental and metabolic contexts."},"narrative":{"mechanistic_narrative":"KCTD10 is the substrate-recognition subunit of a CUL3-RING (CRL3) E3 ubiquitin ligase that selects diverse protein substrates for ubiquitination, thereby controlling endothelial barrier function, cancer cell behavior, brain development, and genome stability [PMID:29358211, PMID:41062692]. Through its BTB domain it bridges CUL3 to substrates and directs distinct ubiquitin linkages to different fates: K63-linked polyubiquitination of RhoB at K162/K181 routes RhoB to lysosomes to govern endothelial contraction and barrier integrity, and the same axis relieves RhoB-mediated suppression of Rac1 to enable membrane ruffling and proliferation [PMID:29358211, PMID:30515933]; K27-linked ubiquitination of EIF3D and K48-linked degradation of β-catenin (via BTB engagement of the Armadillo repeats), the latter lowering PD-L1, are additional substrate routes [PMID:31280863, PMID:40873559]. KCTD10 further destabilizes SLC7A11 to limit cystine uptake and promote ferroptosis, with USP18 acting as the counteracting deubiquitylase [PMID:38959043], and degrades IGF2BP1 to curb m6A-dependent IL-11 mRNA stabilization and immune evasion [PMID:42107984]. In development it degrades KCTD13 to control cortical neuronal progenitor proliferation [PMID:38489388] and degrades Notch1 in concert with Cullin3, with KCTD10 loss causing embryonic lethality with angiogenesis and heart defects and Notch upregulation [PMID:25401743], while it also acts upstream of Notch in brown adipose thermogenesis [PMID:34854382] and directly binds and represses the transcription factor Tbx5a during cardiac development [PMID:24430697]. At co-directional transcription-replication conflicts KCTD10 acts as a bivalent sensor, simultaneously engaging the replisome and transcription machinery and forming higher-order assemblies that recruit CUL3 to ubiquitinate and remove the elongation factor TCEA2, permitting replisome bypass [PMID:41062692]. KCTD10 partitions into condensates through liquid-liquid phase separation driven by its intrinsically disordered region; the schizophrenia-associated p.C124W mutation disrupts this phase separation and impairs RhoB degradation, causing synaptic abnormalities and schizophrenia-like behavior in mice [PMID:39565307].","teleology":[{"year":2005,"claim":"Established the first molecular associations of KCTD10, placing it with the DNA replication/repair machinery before any enzymatic role was known.","evidence":"Cloning and protein interaction assays linking rat KCTD10 to DNA polymerase delta small subunit and PCNA","pmids":["15982757"],"confidence":"Medium","gaps":["No catalytic activity assigned","Interaction with replication machinery not mechanistically connected to a ubiquitination function at this stage"]},{"year":2009,"claim":"Confirmed the PCNA interaction and tied KCTD10 to cell proliferation, with nuclear localization, reinforcing a replication-associated role.","evidence":"Yeast two-hybrid, reciprocal Co-IP, siRNA knockdown proliferation assay and immunofluorescence in A549 cells","pmids":["19125419"],"confidence":"Medium","gaps":["Mechanism linking PCNA binding to proliferation unresolved","No E3 ligase activity demonstrated"]},{"year":2009,"claim":"Defined transcriptional control of KCTD10 itself, showing opposing SP1 activation and AP-2alpha repression at its promoter.","evidence":"Deletion/site-directed mutagenesis, ChIP, luciferase reporter assays, qRT-PCR and western blot","pmids":["19154347"],"confidence":"High","gaps":["Upstream signals controlling SP1/AP-2alpha balance not defined","Does not address KCTD10 protein function"]},{"year":2012,"claim":"Identified KCTD10 as a regulator of NF-kappaB/AP-1 transcriptional activity and a target of TNFAIP1-promoted proteasomal degradation, introducing reciprocal regulation within the KCTD/TNFAIP1 family.","evidence":"Yeast two-hybrid, GST pull-down, Co-IP, MG132 ubiquitination assay, luciferase reporter assays","pmids":["22810651"],"confidence":"Medium","gaps":["E3 ligase responsible for KCTD10 turnover not defined","Direct vs indirect effect on NF-kappaB/AP-1 unresolved"]},{"year":2014,"claim":"Revealed KCTD10's developmental requirement, showing it directly binds and represses Tbx5a to pattern the atrioventricular canal in vivo.","evidence":"Zebrafish mutant screen, morpholino epistasis with tbx5a and has2, direct Kctd10-Tbx5 binding assay, cardiac marker in situ hybridization","pmids":["24430697"],"confidence":"High","gaps":["Whether repression is ubiquitination-dependent not addressed","Relationship to CUL3 complex unclear in this context"]},{"year":2014,"claim":"Connected KCTD10 to the CUL3 E3 machinery and Notch signaling, establishing that it mediates Notch1 degradation and is essential for embryonic angiogenesis and cardiac development.","evidence":"KCTD10 knockout mice (~E10.5 lethality), endogenous Co-IP with Cullin3 and Notch1, qRT-PCR, VEGF stimulation in HUVECs; parallel zebrafish knockdown linking heart defects to RhoA/PCNA","pmids":["25401743","24705121"],"confidence":"Medium","gaps":["Direct ubiquitination of Notch1 not biochemically resolved","Ubiquitin linkage type unspecified"]},{"year":2018,"claim":"Defined the first rigorous substrate-and-linkage mechanism: the CUL3-Rbx1-KCTD10 ligase catalyzes K63 polyubiquitination of RhoB to route it to lysosomes and control endothelial barrier integrity.","evidence":"Co-IP, K63-specific ubiquitination assays, RhoB lysine mutagenesis (K162/K181), lysosomal inhibition, endothelial barrier (TEER/permeability) assays","pmids":["29358211"],"confidence":"High","gaps":["How KCTD10 selects RhoB over other Rho GTPases not defined","In vivo barrier relevance not yet tested at this stage"]},{"year":2019,"claim":"Extended the RhoB axis to cancer signaling and identified a second substrate, showing CUL3/KCTD10-driven RhoB degradation enables Rac1 activation and proliferation, and that the complex K27-ubiquitinates EIF3D.","evidence":"siRNA depletion, Rac1 activity and proliferation assays, membrane ruffle quantification (RhoB/Rac1); MS, K27-specific ubiquitination assay and EIF3D lysine mutagenesis (K153/K275)","pmids":["30515933","31280863"],"confidence":"Medium","gaps":["Functional consequence of EIF3D K27 ubiquitination not established","Substrate determinants distinguishing linkage choice unknown"]},{"year":2022,"claim":"Placed KCTD10 genetically upstream of Notch in metabolic tissue, where it restrains brown-fat thermogenesis.","evidence":"BAT-specific knockdown and overexpression mice, UCP1 western blot, Notch inhibitor rescue, metabolic phenotyping","pmids":["34854382"],"confidence":"Medium","gaps":["Whether the Notch effect is via direct KCTD10-mediated ubiquitination in BAT not shown","Substrate in this context not identified"]},{"year":2024,"claim":"Expanded the substrate repertoire and physiological reach: KCTD10 destabilizes SLC7A11 to promote ferroptosis (opposed by USP18), degrades KCTD13 to control cortical development, and undergoes IDR-driven phase separation required for RhoB degradation at synapses, disrupted by a schizophrenia mutation.","evidence":"MLN4924, ubiquitination/cystine-uptake/ferroptosis assays and tumor growth (SLC7A11); brain-specific KO, Co-IP and ubiquitination, cortical markers and behavior (KCTD13); C124W knockin mice, LLPS and IDR-deletion assays, PSD fractionation (RhoB)","pmids":["38959043","38489388","39565307"],"confidence":"High","gaps":["How phase separation is coupled to E3 catalytic output mechanistically unresolved","Structural basis of distinct substrate recognition across contexts not defined"]},{"year":2025,"claim":"Revealed a genome-stability role and further substrates: KCTD10 senses co-directional transcription-replication conflicts and removes TCEA2 to allow replisome bypass, degrades β-catenin via its BTB domain (lowering PD-L1), and degrades IGF2BP1 to limit m6A-driven IL-11 immune evasion.","evidence":"Co-IP with replisome and transcription machinery, TCEA2 ubiquitination and TRC/DNA-damage assays in KO cells (Nature); IP-MS, K48 ubiquitination, domain mapping and endothelial-specific KO mice (β-catenin); ubiquitination, half-life, RNA pull-down and allograft (IGF2BP1)","pmids":["41062692","40873559","42107984"],"confidence":"High","gaps":["How KCTD10 discriminates co-directional from head-on conflicts mechanistically unclear","Whether higher-order assembly applies to non-TRC substrates unknown"]},{"year":null,"claim":"It remains unresolved how a single substrate adaptor selects such diverse substrates and dictates different ubiquitin linkage types (K27, K48, K63) and outcomes across tissues, and how phase separation, conflict sensing, and CUL3 catalysis are integrated.","evidence":"","pmids":[],"confidence":"High","gaps":["No structural model of KCTD10 substrate recognition across its many targets","Rules governing K27/K48/K63 linkage selection unknown","Relationship between early PCNA/replication interactions and the later TRC-sensing role not biochemically reconciled"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[7,9,11,12,14,15,16]},{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[7,9,14,15]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[7,14,15]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[4]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[1]}],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[7,11,12,15]},{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[14]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[4,5,12]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[5,8,10,15]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[11]}],"complexes":["CUL3-RING (CRL3) E3 ubiquitin ligase"],"partners":["CUL3","RHOB","NOTCH1","SLC7A11","KCTD13","EIF3D","TCEA2","CTNNB1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9H3F6","full_name":"BTB/POZ domain-containing adapter for CUL3-mediated RhoA degradation protein 3","aliases":["BTB/POZ domain-containing protein KCTD10","Potassium channel tetramerization domain-containing protein 10"],"length_aa":313,"mass_kda":35.4,"function":"Substrate-specific adapter of a BCR (BTB-CUL3-RBX1) E3 ubiquitin-protein ligase complex. The BCR(BACURD3) E3 ubiquitin ligase complex mediates the ubiquitination of target proteins, leading to their degradation by the proteasome (By similarity)","subcellular_location":"Nucleus","url":"https://www.uniprot.org/uniprotkb/Q9H3F6/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/KCTD10","classification":"Not Classified","n_dependent_lines":404,"n_total_lines":1208,"dependency_fraction":0.3344370860927152},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/KCTD10","total_profiled":1310},"omim":[{"mim_id":"614950","title":"TRANSMEMBRANE PROTEIN 17; TMEM17","url":"https://www.omim.org/entry/614950"},{"mim_id":"614949","title":"TRANSMEMBRANE PROTEIN 231; TMEM231","url":"https://www.omim.org/entry/614949"},{"mim_id":"614144","title":"B9 DOMAIN-CONTAINING PROTEIN 1; B9D1","url":"https://www.omim.org/entry/614144"},{"mim_id":"613421","title":"POTASSIUM CHANNEL TETRAMERIZATION DOMAIN-CONTAINING PROTEIN 10; KCTD10","url":"https://www.omim.org/entry/613421"},{"mim_id":"612013","title":"COILED-COIL AND C2 DOMAINS-CONTAINING PROTEIN 2A; CC2D2A","url":"https://www.omim.org/entry/612013"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/KCTD10"},"hgnc":{"alias_symbol":["MSTP028","BTBD28"],"prev_symbol":[]},"alphafold":{"accession":"Q9H3F6","domains":[{"cath_id":"3.30.710.10","chopping":"19-128","consensus_level":"high","plddt":90.5843,"start":19,"end":128},{"cath_id":"3.40.30.10","chopping":"141-274","consensus_level":"high","plddt":90.3192,"start":141,"end":274}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9H3F6","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9H3F6-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9H3F6-F1-predicted_aligned_error_v6.png","plddt_mean":82.25},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=KCTD10","jax_strain_url":"https://www.jax.org/strain/search?query=KCTD10"},"sequence":{"accession":"Q9H3F6","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9H3F6.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9H3F6/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9H3F6"}},"corpus_meta":[{"pmid":"38959043","id":"PMC_38959043","title":"The CRL3KCTD10 ubiquitin ligase-USP18 axis coordinately regulates cystine uptake and ferroptosis by modulating SLC7A11.","date":"2024","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/38959043","citation_count":49,"is_preprint":false},{"pmid":"29358211","id":"PMC_29358211","title":"The Cullin-3-Rbx1-KCTD10 complex controls endothelial barrier function via K63 ubiquitination of RhoB.","date":"2018","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/29358211","citation_count":49,"is_preprint":false},{"pmid":"24430697","id":"PMC_24430697","title":"Kctd10 regulates heart morphogenesis by repressing the transcriptional activity of Tbx5a in zebrafish.","date":"2014","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/24430697","citation_count":44,"is_preprint":false},{"pmid":"30515933","id":"PMC_30515933","title":"Cullin-3/KCTD10 E3 complex is essential for Rac1 activation through RhoB degradation in human epidermal growth factor receptor 2-positive breast cancer cells.","date":"2019","source":"Cancer science","url":"https://pubmed.ncbi.nlm.nih.gov/30515933","citation_count":42,"is_preprint":false},{"pmid":"25401743","id":"PMC_25401743","title":"KCTD10 is involved in the cardiovascular system and Notch signaling during early embryonic development.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/25401743","citation_count":33,"is_preprint":false},{"pmid":"15982757","id":"PMC_15982757","title":"A novel PDIP1-related protein, KCTD10, that interacts with proliferating cell nuclear antigen and DNA polymerase delta.","date":"2005","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/15982757","citation_count":31,"is_preprint":false},{"pmid":"19125419","id":"PMC_19125419","title":"KCTD10 interacts with proliferating cell nuclear antigen and its down-regulation could inhibit cell proliferation.","date":"2009","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/19125419","citation_count":30,"is_preprint":false},{"pmid":"19605566","id":"PMC_19605566","title":"Novel variants at KCTD10, MVK, and MMAB genes interact with dietary carbohydrates to modulate HDL-cholesterol concentrations in the Genetics of Lipid Lowering Drugs and Diet Network Study.","date":"2009","source":"The American journal of clinical nutrition","url":"https://pubmed.ncbi.nlm.nih.gov/19605566","citation_count":24,"is_preprint":false},{"pmid":"22810651","id":"PMC_22810651","title":"TNFAIP1 interacts with KCTD10 to promote the degradation of KCTD10 proteins and inhibit the transcriptional activities of NF-κB and AP-1.","date":"2012","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/22810651","citation_count":18,"is_preprint":false},{"pmid":"19154347","id":"PMC_19154347","title":"Transcription factor specificity protein 1 (SP1) and activating protein 2alpha (AP-2alpha) regulate expression of human KCTD10 gene by binding to proximal region of promoter.","date":"2009","source":"The FEBS journal","url":"https://pubmed.ncbi.nlm.nih.gov/19154347","citation_count":17,"is_preprint":false},{"pmid":"24705121","id":"PMC_24705121","title":"KCTD10 is critical for heart and blood vessel development of zebrafish.","date":"2014","source":"Acta biochimica et biophysica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/24705121","citation_count":15,"is_preprint":false},{"pmid":"27716295","id":"PMC_27716295","title":"Association of KCTD10, MVK, and MMAB polymorphisms with dyslipidemia and coronary heart disease in Han Chinese population.","date":"2016","source":"Lipids in health and disease","url":"https://pubmed.ncbi.nlm.nih.gov/27716295","citation_count":14,"is_preprint":false},{"pmid":"31280863","id":"PMC_31280863","title":"Cullin-3/KCTD10 complex is essential for K27-polyubiquitination of EIF3D in human hepatocellular carcinoma HepG2 cells.","date":"2019","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/31280863","citation_count":14,"is_preprint":false},{"pmid":"32484264","id":"PMC_32484264","title":"KCTD10 Biology: An Adaptor for the Ubiquitin E3 Complex Meets Multiple Substrates: Emerging Divergent Roles of the cullin-3/KCTD10 E3 Ubiquitin Ligase Complex in Various Cell Lines.","date":"2020","source":"BioEssays : news and reviews in molecular, cellular and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/32484264","citation_count":13,"is_preprint":false},{"pmid":"30478832","id":"PMC_30478832","title":"Downregulation of microRNA-592 protects mice from hypoplastic heart and congenital heart disease by inhibition of the Notch signaling pathway through upregulating KCTD10.","date":"2018","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/30478832","citation_count":13,"is_preprint":false},{"pmid":"32800877","id":"PMC_32800877","title":"Melanocyte Precursors in the Hair Follicle Bulge of Repigmented Vitiligo Skin Are Controlled by RHO-GTPase, KCTD10, and CTNNB1 Signaling.","date":"2020","source":"The Journal of investigative dermatology","url":"https://pubmed.ncbi.nlm.nih.gov/32800877","citation_count":13,"is_preprint":false},{"pmid":"34854382","id":"PMC_34854382","title":"KCTD10 regulates brown adipose tissue thermogenesis and metabolic function via Notch signaling.","date":"2022","source":"The Journal of endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/34854382","citation_count":12,"is_preprint":false},{"pmid":"31583032","id":"PMC_31583032","title":"Haplotypes of HTRA1 rs1120638, TIMP3 rs9621532, VEGFA rs833068, CFI rs10033900, ERCC6 rs3793784, and KCTD10 rs56209061 Gene Polymorphisms in Age-Related Macular Degeneration.","date":"2019","source":"Disease markers","url":"https://pubmed.ncbi.nlm.nih.gov/31583032","citation_count":10,"is_preprint":false},{"pmid":"36777839","id":"PMC_36777839","title":"KCTD10 functions as a tumor suppressor in hepatocellular carcinoma by triggering the Notch signaling pathway.","date":"2023","source":"American journal of translational research","url":"https://pubmed.ncbi.nlm.nih.gov/36777839","citation_count":9,"is_preprint":false},{"pmid":"38489388","id":"PMC_38489388","title":"KCTD10 regulates brain development by destabilizing brain disorder-associated protein KCTD13.","date":"2024","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/38489388","citation_count":8,"is_preprint":false},{"pmid":"36381427","id":"PMC_36381427","title":"Inhibition of KCTD10 Affects Diabetic Retinopathy Progression by Reducing VEGF and Affecting Angiogenesis.","date":"2022","source":"Genetics 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immunology","url":"https://pubmed.ncbi.nlm.nih.gov/40873559","citation_count":3,"is_preprint":false},{"pmid":"18257228","id":"PMC_18257228","title":"[Preparation of mouse KCTD10 antibody and expression analysis of KCTD10 in neuroepithelium of neural tube and dorsal root ganglion].","date":"2007","source":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/18257228","citation_count":3,"is_preprint":false},{"pmid":"40059128","id":"PMC_40059128","title":"Elucidating the role of KCTD10 in coronary atherosclerosis: Harnessing bioinformatics and machine learning to advance understanding.","date":"2025","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/40059128","citation_count":2,"is_preprint":false},{"pmid":"40121532","id":"PMC_40121532","title":"Identification of de novo variants in KCTD10 as a proposed cause for multiple congenital anomalies.","date":"2025","source":"HGG advances","url":"https://pubmed.ncbi.nlm.nih.gov/40121532","citation_count":1,"is_preprint":false},{"pmid":"40795335","id":"PMC_40795335","title":"KCTD10 promoting PD-L1 expression in colorectal cancer enhanced the anti-tumor effect of PD-1 antibody.","date":"2025","source":"Integrative biology : quantitative biosciences from nano to macro","url":"https://pubmed.ncbi.nlm.nih.gov/40795335","citation_count":0,"is_preprint":false},{"pmid":"42128708","id":"PMC_42128708","title":"KCTD10 resolves co-directional transcription-replication conflicts.","date":"2026","source":"Trends in cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/42128708","citation_count":0,"is_preprint":false},{"pmid":"40634059","id":"PMC_40634059","title":"Nicotine Induces Fetal Cardiac Dysfunction by Promoting Cardiomyocyte Apoptosis Through Regulating the KCTD10-Notch Signaling.","date":"2025","source":"International heart journal","url":"https://pubmed.ncbi.nlm.nih.gov/40634059","citation_count":0,"is_preprint":false},{"pmid":"42107984","id":"PMC_42107984","title":"KCTD10 weakens m6A modification of IL-11 in a ubiquitin-dependent manner to repress tumour immune escape in lung adenocarcinoma.","date":"2026","source":"British journal of pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/42107984","citation_count":0,"is_preprint":false},{"pmid":"42037670","id":"PMC_42037670","title":"KCTD10 as a selective cancer dependency from transcription-replication conflicts (TRCs).","date":"2026","source":"Ageing and cancer research & treatment","url":"https://pubmed.ncbi.nlm.nih.gov/42037670","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":16906,"output_tokens":4445,"usd":0.058696,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":12311,"output_tokens":4490,"usd":0.086902,"stage2_stop_reason":"end_turn"},"total_usd":0.145598,"stage1_batch_id":"msgbatch_01BbTCMyrRPyhtEq6b1MdwCN","stage2_batch_id":"msgbatch_011dEed5bbxYvTAaQWKJevxu","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2005,\n      \"finding\": \"KCTD10 (rat) interacts with the small subunit of DNA polymerase delta and PCNA, identifying it as a member of the PDIP1 gene family involved in DNA replication/repair.\",\n      \"method\": \"Protein interaction assays (by analogy with PDIP1 family characterization; cloning and expression analysis)\",\n      \"journal\": \"Biochimica et biophysica acta\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — initial cloning paper with interaction data, replicated in subsequent studies\",\n      \"pmids\": [\"15982757\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"KCTD10 interacts with PCNA (confirmed by yeast two-hybrid and co-immunoprecipitation), and its knockdown inhibits cell proliferation in A549 cells; nuclear localization was observed.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation, siRNA knockdown with cell proliferation assay, immunofluorescence\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — reciprocal Co-IP plus functional KD assay, single lab\",\n      \"pmids\": [\"19125419\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"SP1 binding to the proximal KCTD10 promoter stimulates KCTD10 transcription, while AP-2alpha binding to the same region represses it, as established by deletion mutagenesis, site-directed mutation, ChIP, and reporter assays.\",\n      \"method\": \"Deletion mutagenesis, site-directed mutagenesis, chromatin immunoprecipitation (ChIP), luciferase reporter assay, qRT-PCR, western blot\",\n      \"journal\": \"The FEBS journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (ChIP, mutagenesis, reporter assay) in single lab with rigorous controls\",\n      \"pmids\": [\"19154347\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"TNFAIP1 interacts with KCTD10 and promotes ubiquitin-mediated proteasomal degradation of KCTD10 protein; both KCTD10 and TNFAIP1 inhibit NF-κB and AP-1 transcriptional activity.\",\n      \"method\": \"Yeast two-hybrid, GST pull-down, co-immunoprecipitation, colocalization, protein degradation assay, ubiquitination assay with MG132, luciferase reporter assay\",\n      \"journal\": \"Molecular biology reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (Y2H, GST pull-down, Co-IP, ubiquitination assay), single lab\",\n      \"pmids\": [\"22810651\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"In zebrafish, Kctd10 directly binds Tbx5a (T-box transcription factor) and represses its transcriptional activity; loss of kctd10 causes atrioventricular canal defects rescued by tbx5a or has2 knockdown.\",\n      \"method\": \"Zebrafish mutant genetic screen, morpholino knockdown epistasis, protein binding assay (Kctd10-Tbx5 direct binding), in situ hybridization for cardiac markers\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis in vivo plus direct protein binding assay, multiple orthogonal approaches\",\n      \"pmids\": [\"24430697\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"KCTD10 interacts with Cullin3 and Notch1 simultaneously, mediating Notch1 proteolytic degradation; KCTD10 knockout mice die ~E10.5 with severe angiogenesis and heart defects, and Notch signaling members are upregulated in KCTD10-deficient embryos and KCTD10-silenced HUVECs.\",\n      \"method\": \"KCTD10 knockout mouse generation, co-immunoprecipitation (endogenous), qRT-PCR, western blotting, VEGF stimulation assays\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — endogenous Co-IP plus in vivo KO phenotype, single lab\",\n      \"pmids\": [\"25401743\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Zebrafish kctd10 knockdown causes pericardial edema, loss of heart formation, and loss of intersomitic vessels; the heart defect is linked to RhoA and PCNA pathways.\",\n      \"method\": \"Morpholino knockdown in zebrafish, in situ hybridization for cardiac markers (amhc, vmhc, cmlc2), Flk-1 staining for vasculature\",\n      \"journal\": \"Acta biochimica et biophysica Sinica\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KD with defined cellular/developmental phenotype and marker analysis, single lab\",\n      \"pmids\": [\"24705121\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"The Cullin-3-Rbx1-KCTD10 E3 ligase complex catalyzes K63 polyubiquitination of RhoB at lysine 162 and 181, targeting RhoB to lysosomes and thereby regulating endothelial barrier integrity via control of RhoB-mediated endothelial cell contraction.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays (K63-specific), site-directed mutagenesis of RhoB lysines, lysosomal pathway inhibition, siRNA knockdown with endothelial barrier assay (TEER/permeability)\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro ubiquitination assay with mutagenesis, Co-IP, functional barrier assay, multiple orthogonal methods\",\n      \"pmids\": [\"29358211\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"The CUL3/KCTD10 E3 ligase ubiquitinates RhoB for degradation, which relieves RhoB-mediated inhibition of Rac1 at the plasma membrane, enabling EGF-induced Rac1 activation, dorsal membrane ruffle formation, and cell proliferation in HER2-positive breast cancer cells.\",\n      \"method\": \"siRNA depletion of CUL3 or KCTD10, Rac1 activity assay, cell proliferation assay, membrane ruffle quantification, co-immunoprecipitation\",\n      \"journal\": \"Cancer science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional epistasis (CUL3/KCTD10 → RhoB → Rac1) with multiple assays, single lab\",\n      \"pmids\": [\"30515933\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"The CUL3/KCTD10 complex forms a complex with EIF3D and catalyzes K27 polyubiquitination of EIF3D at lysine 153 and 275 residues in hepatocellular carcinoma HepG2 cells.\",\n      \"method\": \"Mass spectrometry (complex identification), co-immunoprecipitation, ubiquitination assay with K27-specific linkage, site-directed mutagenesis of EIF3D lysines, KCTD10 depletion\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MS identification plus mutagenesis-confirmed ubiquitination sites, single lab\",\n      \"pmids\": [\"31280863\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"KCTD10 acts as an upstream regulator of Notch signaling in brown adipose tissue; KCTD10 overexpression suppresses UCP1 expression and thermogenesis, while BAT-specific KCTD10 knockdown increases thermogenesis and cold tolerance; inhibiting Notch signaling restores KCTD10-overexpression-suppressed thermogenesis, placing KCTD10 upstream of Notch.\",\n      \"method\": \"BAT-specific knockdown mouse model, KCTD10 overexpression in BAT, UCP1 western blot, Notch inhibitor rescue experiment, metabolic phenotyping\",\n      \"journal\": \"The Journal of endocrinology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis in vivo (Notch inhibitor rescue) plus KD/OE with defined phenotype, single lab\",\n      \"pmids\": [\"34854382\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"KCTD10 is the substrate-recognizing subunit of CRL3 E3 ligase that ubiquitinates SLC7A11 (cystine transporter); USP18 is the counteracting deubiquitylase. Upon cystine deprivation, KCTD10 protein decreases and USP18 increases, causing SLC7A11 accumulation. KCTD10 destabilizes SLC7A11 to reduce cystine uptake and promote ferroptosis.\",\n      \"method\": \"neddylation inhibitor (MLN4924) treatment, KCTD10 and USP18 siRNA/overexpression, ubiquitination assays, cystine uptake assay, ferroptosis assays, co-immunoprecipitation, in vivo tumor growth assay\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (Co-IP, ubiquitination assay, functional cystine/ferroptosis assays, in vivo), rigorous mechanistic dissection\",\n      \"pmids\": [\"38959043\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"KCTD10 mediates ubiquitination-dependent degradation of KCTD13 protein in the developing cortex; KCTD10 deficiency causes KCTD13 accumulation, abnormal neuronal progenitor proliferation/differentiation, reduced deep-layer neurons, and motor deficits in brain-specific Kctd10 KO mice.\",\n      \"method\": \"Brain-specific Kctd10 knockout mice, co-immunoprecipitation (KCTD10-KCTD13 interaction screen), ubiquitination assay, cortical layer marker analysis, behavioral testing\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — in vivo KO with defined phenotype, Co-IP interaction, ubiquitination assay, multiple orthogonal methods in single rigorous study\",\n      \"pmids\": [\"38489388\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"KCTD10 undergoes liquid-liquid phase separation (LLPS) mediated by its intrinsically disordered region (IDR); the schizophrenia-associated p.C124W mutation disrupts LLPS, impairing RHOB degradation and causing excessive RHOB accumulation in postsynaptic density, leading to synaptic abnormalities and schizophrenia-like behaviors in heterozygous mice.\",\n      \"method\": \"Heterozygous KCTD10 C124W knockin mice, LLPS assay, IDR deletion mutant, ubiquitination/degradation assay for RHOB, postsynaptic density fractionation, behavioral assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — in vivo knockin mice, LLPS mechanistic assays, domain mutant analysis, multiple orthogonal methods in single rigorous study\",\n      \"pmids\": [\"39565307\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"The CUL3-KCTD10 E3 ligase acts as a bivalent sensor of co-directional transcription-replication conflicts (TRCs): KCTD10 interacts with both the replisome and transcription machinery simultaneously; upon detecting co-directional TRCs, KCTD10 complexes assemble higher-order assemblies recruiting CUL3 to ubiquitinate and remove the RNA polymerase elongation factor TCEA2, allowing replisome bypass. KCTD10 loss causes TCEA2 retention, TRC accumulation, and increased DNA damage.\",\n      \"method\": \"Co-immunoprecipitation (KCTD10 with replisome and transcription machinery), ubiquitination assay for TCEA2, TRC detection assay, DNA damage markers, KCTD10 KO cells\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (Co-IP with both complexes, ubiquitination assay, TRC assay, KO phenotype), peer-reviewed in Nature\",\n      \"pmids\": [\"41062692\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"The BTB domain of KCTD10 interacts with Armadillo repeat domains 1-9 of β-catenin and facilitates K48-linked ubiquitin-dependent degradation of β-catenin, leading to downregulation of PD-L1; endothelial-specific Kctd10 knockout promotes lung cancer metastasis and tumor angiogenesis via β-catenin signaling.\",\n      \"method\": \"IP-mass spectrometry, co-immunoprecipitation, ubiquitination assay (K48-specific), domain mapping, Kctd10 endothelial-specific KO mice (Kctd10flox/flox CDH5-CreERT2), in vivo tumor assays\",\n      \"journal\": \"Frontiers in immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — IP-MS plus domain-mapping Co-IP, K48-specific ubiquitination assay, conditional KO in vivo, multiple orthogonal methods\",\n      \"pmids\": [\"40873559\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"KCTD10 promotes ubiquitination and proteasomal degradation of IGF2BP1; KCTD10 overexpression reduces IGF2BP1-mediated m6A stabilization of IL-11 mRNA, thereby restraining immune evasion in lung adenocarcinoma.\",\n      \"method\": \"Ubiquitination assay, IGF2BP1 half-life detection, RNA pull-down (IGF2BP1-IL-11 mRNA), ELISA, western blot, co-immunoprecipitation, allograft experiment\",\n      \"journal\": \"British journal of pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ubiquitination assay plus functional downstream readout (m6A/IL-11 axis), single lab with multiple methods\",\n      \"pmids\": [\"42107984\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"KCTD10 functions primarily as the substrate-recognition adaptor of the CUL3-RING E3 ubiquitin ligase complex, targeting diverse substrates (RhoB via K63-ubiquitination for lysosomal degradation, SLC7A11 for proteasomal degradation, EIF3D via K27-ubiquitination, KCTD13 for degradation, β-catenin via K48-ubiquitination, IGF2BP1, and the RNA Pol II elongation factor TCEA2) to regulate endothelial barrier function, ferroptosis, cancer cell proliferation, brain development, and genome stability at transcription-replication conflicts; it also undergoes liquid-liquid phase separation via its intrinsically disordered region to facilitate RhoB degradation at synapses, and can directly bind and repress transcription factors (Tbx5a) to control cardiac development, while negatively regulating Notch signaling in multiple developmental and metabolic contexts.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"KCTD10 is the substrate-recognition subunit of a CUL3-RING (CRL3) E3 ubiquitin ligase that selects diverse protein substrates for ubiquitination, thereby controlling endothelial barrier function, cancer cell behavior, brain development, and genome stability [#7, #14]. Through its BTB domain it bridges CUL3 to substrates and directs distinct ubiquitin linkages to different fates: K63-linked polyubiquitination of RhoB at K162/K181 routes RhoB to lysosomes to govern endothelial contraction and barrier integrity, and the same axis relieves RhoB-mediated suppression of Rac1 to enable membrane ruffling and proliferation [#7, #8]; K27-linked ubiquitination of EIF3D and K48-linked degradation of β-catenin (via BTB engagement of the Armadillo repeats), the latter lowering PD-L1, are additional substrate routes [#9, #15]. KCTD10 further destabilizes SLC7A11 to limit cystine uptake and promote ferroptosis, with USP18 acting as the counteracting deubiquitylase [#11], and degrades IGF2BP1 to curb m6A-dependent IL-11 mRNA stabilization and immune evasion [#16]. In development it degrades KCTD13 to control cortical neuronal progenitor proliferation [#12] and degrades Notch1 in concert with Cullin3, with KCTD10 loss causing embryonic lethality with angiogenesis and heart defects and Notch upregulation [#5], while it also acts upstream of Notch in brown adipose thermogenesis [#10] and directly binds and represses the transcription factor Tbx5a during cardiac development [#4]. At co-directional transcription-replication conflicts KCTD10 acts as a bivalent sensor, simultaneously engaging the replisome and transcription machinery and forming higher-order assemblies that recruit CUL3 to ubiquitinate and remove the elongation factor TCEA2, permitting replisome bypass [#14]. KCTD10 partitions into condensates through liquid-liquid phase separation driven by its intrinsically disordered region; the schizophrenia-associated p.C124W mutation disrupts this phase separation and impairs RhoB degradation, causing synaptic abnormalities and schizophrenia-like behavior in mice [#13].\",\n  \"teleology\": [\n    {\n      \"year\": 2005,\n      \"claim\": \"Established the first molecular associations of KCTD10, placing it with the DNA replication/repair machinery before any enzymatic role was known.\",\n      \"evidence\": \"Cloning and protein interaction assays linking rat KCTD10 to DNA polymerase delta small subunit and PCNA\",\n      \"pmids\": [\"15982757\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No catalytic activity assigned\", \"Interaction with replication machinery not mechanistically connected to a ubiquitination function at this stage\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Confirmed the PCNA interaction and tied KCTD10 to cell proliferation, with nuclear localization, reinforcing a replication-associated role.\",\n      \"evidence\": \"Yeast two-hybrid, reciprocal Co-IP, siRNA knockdown proliferation assay and immunofluorescence in A549 cells\",\n      \"pmids\": [\"19125419\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking PCNA binding to proliferation unresolved\", \"No E3 ligase activity demonstrated\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defined transcriptional control of KCTD10 itself, showing opposing SP1 activation and AP-2alpha repression at its promoter.\",\n      \"evidence\": \"Deletion/site-directed mutagenesis, ChIP, luciferase reporter assays, qRT-PCR and western blot\",\n      \"pmids\": [\"19154347\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Upstream signals controlling SP1/AP-2alpha balance not defined\", \"Does not address KCTD10 protein function\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Identified KCTD10 as a regulator of NF-kappaB/AP-1 transcriptional activity and a target of TNFAIP1-promoted proteasomal degradation, introducing reciprocal regulation within the KCTD/TNFAIP1 family.\",\n      \"evidence\": \"Yeast two-hybrid, GST pull-down, Co-IP, MG132 ubiquitination assay, luciferase reporter assays\",\n      \"pmids\": [\"22810651\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"E3 ligase responsible for KCTD10 turnover not defined\", \"Direct vs indirect effect on NF-kappaB/AP-1 unresolved\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Revealed KCTD10's developmental requirement, showing it directly binds and represses Tbx5a to pattern the atrioventricular canal in vivo.\",\n      \"evidence\": \"Zebrafish mutant screen, morpholino epistasis with tbx5a and has2, direct Kctd10-Tbx5 binding assay, cardiac marker in situ hybridization\",\n      \"pmids\": [\"24430697\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether repression is ubiquitination-dependent not addressed\", \"Relationship to CUL3 complex unclear in this context\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Connected KCTD10 to the CUL3 E3 machinery and Notch signaling, establishing that it mediates Notch1 degradation and is essential for embryonic angiogenesis and cardiac development.\",\n      \"evidence\": \"KCTD10 knockout mice (~E10.5 lethality), endogenous Co-IP with Cullin3 and Notch1, qRT-PCR, VEGF stimulation in HUVECs; parallel zebrafish knockdown linking heart defects to RhoA/PCNA\",\n      \"pmids\": [\"25401743\", \"24705121\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct ubiquitination of Notch1 not biochemically resolved\", \"Ubiquitin linkage type unspecified\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Defined the first rigorous substrate-and-linkage mechanism: the CUL3-Rbx1-KCTD10 ligase catalyzes K63 polyubiquitination of RhoB to route it to lysosomes and control endothelial barrier integrity.\",\n      \"evidence\": \"Co-IP, K63-specific ubiquitination assays, RhoB lysine mutagenesis (K162/K181), lysosomal inhibition, endothelial barrier (TEER/permeability) assays\",\n      \"pmids\": [\"29358211\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How KCTD10 selects RhoB over other Rho GTPases not defined\", \"In vivo barrier relevance not yet tested at this stage\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Extended the RhoB axis to cancer signaling and identified a second substrate, showing CUL3/KCTD10-driven RhoB degradation enables Rac1 activation and proliferation, and that the complex K27-ubiquitinates EIF3D.\",\n      \"evidence\": \"siRNA depletion, Rac1 activity and proliferation assays, membrane ruffle quantification (RhoB/Rac1); MS, K27-specific ubiquitination assay and EIF3D lysine mutagenesis (K153/K275)\",\n      \"pmids\": [\"30515933\", \"31280863\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of EIF3D K27 ubiquitination not established\", \"Substrate determinants distinguishing linkage choice unknown\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Placed KCTD10 genetically upstream of Notch in metabolic tissue, where it restrains brown-fat thermogenesis.\",\n      \"evidence\": \"BAT-specific knockdown and overexpression mice, UCP1 western blot, Notch inhibitor rescue, metabolic phenotyping\",\n      \"pmids\": [\"34854382\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether the Notch effect is via direct KCTD10-mediated ubiquitination in BAT not shown\", \"Substrate in this context not identified\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Expanded the substrate repertoire and physiological reach: KCTD10 destabilizes SLC7A11 to promote ferroptosis (opposed by USP18), degrades KCTD13 to control cortical development, and undergoes IDR-driven phase separation required for RhoB degradation at synapses, disrupted by a schizophrenia mutation.\",\n      \"evidence\": \"MLN4924, ubiquitination/cystine-uptake/ferroptosis assays and tumor growth (SLC7A11); brain-specific KO, Co-IP and ubiquitination, cortical markers and behavior (KCTD13); C124W knockin mice, LLPS and IDR-deletion assays, PSD fractionation (RhoB)\",\n      \"pmids\": [\"38959043\", \"38489388\", \"39565307\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How phase separation is coupled to E3 catalytic output mechanistically unresolved\", \"Structural basis of distinct substrate recognition across contexts not defined\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Revealed a genome-stability role and further substrates: KCTD10 senses co-directional transcription-replication conflicts and removes TCEA2 to allow replisome bypass, degrades β-catenin via its BTB domain (lowering PD-L1), and degrades IGF2BP1 to limit m6A-driven IL-11 immune evasion.\",\n      \"evidence\": \"Co-IP with replisome and transcription machinery, TCEA2 ubiquitination and TRC/DNA-damage assays in KO cells (Nature); IP-MS, K48 ubiquitination, domain mapping and endothelial-specific KO mice (β-catenin); ubiquitination, half-life, RNA pull-down and allograft (IGF2BP1)\",\n      \"pmids\": [\"41062692\", \"40873559\", \"42107984\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How KCTD10 discriminates co-directional from head-on conflicts mechanistically unclear\", \"Whether higher-order assembly applies to non-TRC substrates unknown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how a single substrate adaptor selects such diverse substrates and dictates different ubiquitin linkage types (K27, K48, K63) and outcomes across tissues, and how phase separation, conflict sensing, and CUL3 catalysis are integrated.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No structural model of KCTD10 substrate recognition across its many targets\", \"Rules governing K27/K48/K63 linkage selection unknown\", \"Relationship between early PCNA/replication interactions and the later TRC-sensing role not biochemically reconciled\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [7, 9, 11, 12, 14, 15, 16]},\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [7, 9, 14, 15]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [7, 14, 15]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [7, 11, 12, 15]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [14]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [4, 5, 12]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [5, 8, 10, 15]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [11]}\n    ],\n    \"complexes\": [\n      \"CUL3-RING (CRL3) E3 ubiquitin ligase\"\n    ],\n    \"partners\": [\n      \"CUL3\",\n      \"RhoB\",\n      \"Notch1\",\n      \"SLC7A11\",\n      \"KCTD13\",\n      \"EIF3D\",\n      \"TCEA2\",\n      \"CTNNB1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":6,"faith_pct":83.33333333333333}}