{"gene":"USP24","run_date":"2026-06-11T09:02:06","timeline":{"discoveries":[{"year":2015,"finding":"USP24 directly deubiquitinates p53, stabilizing it in response to UV-induced DNA damage; purified USP24 protein cleaves ubiquitinated p53 in vitro, and USP24 depletion destabilizes p53, impairs PUMA activation, and renders cells resistant to apoptosis after UV irradiation.","method":"In vitro deubiquitination assay with purified USP24; siRNA knockdown with p53 stability and apoptosis readouts; forced re-expression rescue experiment","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with purified protein plus cellular knockdown/rescue with multiple orthogonal readouts in a single focused study","pmids":["25578727"],"is_preprint":false},{"year":2012,"finding":"USP24 interacts with DDB2 (a nucleotide excision repair factor) and stabilizes it by removing ubiquitin; USP24 knockdown decreases steady-state DDB2 levels, and purified USP24 cleaves ubiquitinated DDB2 in vitro.","method":"Yeast two-hybrid screen; co-immunoprecipitation; siRNA knockdown; in vitro deubiquitination assay","journal":"Cell cycle (Georgetown, Tex.)","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution plus reciprocal interaction confirmation and knockdown, single lab with multiple orthogonal methods","pmids":["23159851"],"is_preprint":false},{"year":2019,"finding":"USP24 negatively regulates autophagy by controlling ubiquitination and stability of ULK1; USP24 knockdown in cell lines and iPSC-derived dopaminergic neurons elevates ULK1 protein levels and increases autophagy flux in an MTORC1-independent, class III PtdIns3K-dependent manner.","method":"siRNA knockdown in cell lines and iPSC-derived dopaminergic neurons; autophagy flux assays; MTORC1 and PtdIns3K inhibitor epistasis","journal":"Autophagy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean knockdown with defined cellular phenotype and pathway placement via pharmacological epistasis, single lab","pmids":["30957634"],"is_preprint":false},{"year":2018,"finding":"USP24 stabilizes p300 and β-TrCP proteins in M2 macrophages and lung cancer cells, leading to increased histone-3 acetylation and NF-κB levels while decreasing DNMT1 and IκB, thereby increasing IL-6 transcription.","method":"siRNA knockdown; western blot for substrate levels; conditioned medium transfer assays; IL-6 rescue experiments","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — knockdown with defined molecular mechanism across two cell types, multiple downstream readouts, single lab","pmids":["30266897"],"is_preprint":false},{"year":2021,"finding":"USP24 interacts with GSDMB and prevents its proteasomal degradation (deubiquitination-mediated stabilization), and the USP24/GSDMB complex promotes STAT3 phosphorylation and activation in bladder cancer cells.","method":"Mass spectrometry; co-immunoprecipitation; western blot; USP24 inhibitor treatment","journal":"International journal of biological sciences","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-IP confirmed interaction and stabilization across multiple experiments, single lab","pmids":["34326684"],"is_preprint":false},{"year":2019,"finding":"USP24 stabilizes Mcl-1; WP1130 inhibits USP24 catalytic activity (molecular docking and cellular thermal shift assay), leading to Mcl-1 destabilization and apoptosis via mitochondrial transmembrane potential collapse in T-ALL cells.","method":"Molecular docking; cellular thermal shift assay (CETSA); siRNA knockdown; Mcl-1 protein level measurement; mitochondrial transmembrane potential assay","journal":"Cancer cell international","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CETSA confirms direct drug-target engagement, knockdown phenocopies drug effect, single lab","pmids":["30911287"],"is_preprint":false},{"year":2020,"finding":"USP24 interacts with the bromodomain (BRD) via its C-terminus and stabilizes most BRD-containing proteins by removing ubiquitin from Lys391/Lys400 of the BRD; loss of the USP24-BRD interaction motif abolishes this stabilization and USP24's effects on cancer progression.","method":"Co-immunoprecipitation; western blot for BRD protein stability; mutagenesis of ubiquitin acceptor sites (Lys391/Lys400); deletion mutant analysis","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — site-directed mutagenesis of substrate ubiquitination sites plus interaction mapping, single lab","pmids":["33257797"],"is_preprint":false},{"year":2020,"finding":"USP24 interacts with and stabilizes CRMP2; deletion of Usp24 in mice leads to CRMP2 degradation, causing spindle defects, chromosome missegregation, and aneuploidy that are rescued by CRMP2 restoration, identifying USP24 as a tumor suppressor maintaining mitotic accuracy.","method":"Usp24 knockout mouse model; genetic rescue with CRMP2 restoration; chromosome segregation and spindle defect assays","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic rescue experiment in murine knockout model with defined cellular phenotype, single lab","pmids":["33355202"],"is_preprint":false},{"year":2021,"finding":"USP24 represses DNA-damage repair by decreasing Rad51 expression (promoting genomic instability) and increases ABC transporter levels (P-gp, ABCG2, ezrin) to enhance Taxol efflux from cancer cells, thereby promoting drug resistance.","method":"USP24 knockdown/overexpression; western blot for Rad51 and ABC transporter levels; drug efflux assays; NCI677397 inhibitor treatment","journal":"Cell death and differentiation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — knockdown with defined molecular readouts and pharmacological validation, single lab","pmids":["33846536"],"is_preprint":false},{"year":2023,"finding":"USP24 is recruited to phospho-Runx2 (pRunx2, phosphorylated at Ser28 by p38 kinase) and stabilizes pRunx2 by deubiquitination, allowing pRunx2 to assemble a NCOA3-p300-pRunx2 complex that transactivates ADAMTS genes and promotes ECM degradation in intervertebral disc degeneration.","method":"Co-immunoprecipitation; mass spectrometry; western blot; RT-qPCR; p38 inhibitor epistasis; LPS-induced chronic inflammation mouse model","journal":"Biology direct","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP/MS confirmed complex, pharmacological epistasis defined pathway, single lab","pmids":["37415159"],"is_preprint":false},{"year":2024,"finding":"USP24 reduces K48-linked ubiquitination of Beclin1, thereby stabilizing it and promoting autophagy-dependent ferroptosis in hepatocellular carcinoma; effects of USP24 overexpression on HCC proliferation are partially reversed by Beclin1 silencing.","method":"Co-immunoprecipitation confirming endogenous USP24-Beclin1 interaction; ubiquitination assay (K48-linkage specific); genetic rescue with Beclin1 silencing; in vivo xenograft","journal":"Communications biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus K48-specific ubiquitination assay and genetic epistasis, single lab","pmids":["39379617"],"is_preprint":false},{"year":2024,"finding":"USP24 binds to and deubiquitinates TRAF2, preventing its degradation; stabilized TRAF2 activates the AKT/NF-κB signaling pathway to promote HCC cell survival.","method":"Co-immunoprecipitation; western blot for TRAF2 stability after USP24 manipulation; AKT/NF-κB pathway readouts","journal":"Biochemical pharmacology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-IP and substrate stabilization confirmed across multiple experiments, single lab","pmids":["39127151"],"is_preprint":false},{"year":2024,"finding":"USP24 activates autophagy in both interphase and mitotic periods of lung cancer cells by inhibiting E2F4 and TRAF6, respectively; the catalytic mutant USP24C1695A abolishes this effect, and autophagy induction is required for USP24 inhibitor-mediated suppression of drug resistance and maintenance of genomic integrity and PD-L1 degradation.","method":"CRISPR-mediated catalytic knockout (USP24C1695A); USP24 inhibitor USP24-i-101; bafilomycin-A1 autophagy inhibition rescue; doxycycline-inducible EGFRL858R lung cancer mouse model","journal":"Cell death and differentiation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — catalytic mutant confirms enzyme-dependent mechanism, in vivo mouse model, autophagy epistasis, single lab","pmids":["38491202"],"is_preprint":false},{"year":2025,"finding":"USP24 removes K48-linked polyubiquitin from PD-1, stabilizing PD-1 protein in CD8+ T cells; IL-6 transcriptionally activates USP24 expression, linking the IL-6/STAT3 axis to PD-1 stabilization; USP24 catalytic-deficient (Usp24 catalytic mutant) mice show reduced PD-1 levels and attenuated lung tumorigenesis.","method":"Co-immunoprecipitation; K48-ubiquitination assay; catalytic-deficient mouse model; USP24-i-101 inhibitor treatment; anti-CTLA4 combination in vivo","journal":"Science advances","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — specific ubiquitin linkage identified, in vivo genetic catalytic-mutant model, pharmacological validation, multiple orthogonal methods in one study","pmids":["40238877"],"is_preprint":false},{"year":2025,"finding":"USP24 directly interacts with and deubiquitinates DHODH, stabilizing it to maintain coenzyme Q reduction and protect TNBC cells from lipid peroxidation-driven ferroptosis; pharmacological USP24 inhibition synergizes with ferroptosis inducers via a DHODH-dependent pathway.","method":"Co-immunoprecipitation; deubiquitination assay; siRNA knockdown; in vitro and in vivo ferroptosis assays with DHODH rescue","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus DHODH-dependent genetic rescue and pharmacological validation, single lab","pmids":["40715045"],"is_preprint":false},{"year":2025,"finding":"USP24 stabilizes YAP1 by directly interacting with and deubiquitinating it; USP24 depletion suppresses HCC cell proliferation and tumor growth, and these effects are rescued by YAP1 restoration.","method":"Co-immunoprecipitation; deubiquitination assay; siRNA knockdown; YAP1 rescue experiment; xenograft mouse model","journal":"Cancer cell international","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, deubiquitination assay, and genetic rescue in vivo, single lab","pmids":["40287768"],"is_preprint":false},{"year":2025,"finding":"USP24 interacts with and stabilizes PKA-Cα by deubiquitination; USP24 expression is upregulated by PKA activation during adipocyte differentiation, creating a positive feedback loop that increases CREB phosphorylation and lipogenic gene expression; free fatty acids increase USP24 to activate NF-κB and TGFβ pathways driving inflammation and fibrosis.","method":"Co-immunoprecipitation; CRISPR/Cas9 catalytic knockout (USP24C1695A) mice; USP24-i-101 inhibitor; western blot; RNA-sequencing","journal":"Journal of biomedical science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus catalytic knockout mouse model and transcriptomic data, single lab","pmids":["40448065"],"is_preprint":false},{"year":2026,"finding":"USP24 stabilizes STAT2 through deubiquitination in osteoblasts; loss of USP24 reduces osteoblast differentiation and bone formation, phenocopying STAT2 deficiency; rAAV-mediated Usp24 silencing in a FOP mouse model diminishes heterotopic ossification and reduces STAT2 protein levels.","method":"Co-immunoprecipitation; deubiquitination assay; USP24 knockout/knockdown; STAT2 knockdown phenocopy; rAAV-mediated silencing in FOP mouse model","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic interaction confirmed by phenocopy and in vivo model, single lab","pmids":["42218160"],"is_preprint":false},{"year":2026,"finding":"USP24 deubiquitinates TRAF6, stabilizing it; stabilized TRAF6 then ubiquitinates Beclin-1 to promote its degradation, thereby inhibiting autophagy in diabetic myocardial disorder; TRAF6 overexpression reverses the autophagy rescue caused by USP24 knockdown.","method":"Co-immunoprecipitation; western blot for TRAF6 and Beclin-1 levels; lentiviral knockdown/overexpression; autophagic flux assays (mRFP-GFP-LC3, TEM); genetic epistasis (TRAF6 OE reverses USP24 KD effect)","journal":"Life sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, substrate ubiquitination, and genetic epistasis, single lab","pmids":["41785730"],"is_preprint":false},{"year":2024,"finding":"USP24 is an ISG15 cross-reactive deubiquitylase; recombinant USP24 processes pro-ISG15 and cleaves ISG15-linked synthetic substrates in vitro; USP24 depletion increases ISG15 conjugates upon IFN-β stimulation; USP24 specifically deISGylates the RNA helicase MOV10, and ISGylated MOV10 enhances IFN-β production, while USP24-mediated deISGylation of MOV10 negatively regulates the innate immune response.","method":"Activity-based protein profiling (ABPP); in vitro deISGylation assay with recombinant USP24 and synthetic ISG15-substrates; USP24 depletion; total proteome, GG-peptidome, and ISG15 interactome proteomics","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — in vitro reconstitution with recombinant protein plus proteomics, but preprint not yet peer-reviewed, single study","pmids":[],"is_preprint":true},{"year":2023,"finding":"USP24 promotes NF-κB accumulation in diabetic cardiomyopathy; USP24 knockdown reduces NF-κB levels, whereas NF-κB knockdown does not reduce USP24 expression, establishing USP24 as upstream of NF-κB; elevated NF-κB promotes FACL4 expression and ferroptosis while reducing SLC7A11 and FTH1.","method":"siRNA knockdown in H9c2 cells; PMA-mediated NF-κB re-activation rescue; western blot; in vivo HFD/STZ mouse model","journal":"Free radical biology & medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — directional epistasis established by unidirectional knockdown and pharmacological rescue, single lab","pmids":["38056575"],"is_preprint":false},{"year":2023,"finding":"USP24 interacts with SIRT7 and mediates its deubiquitination; inhibition of miR-21-5p downregulates the ubiquitination level of SIRT7 via USP24, affecting autophagy levels in hepatocellular carcinoma.","method":"Co-immunoprecipitation; western blot for SIRT7 ubiquitination","journal":"Life sciences","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single co-IP experiment, mechanistic link to USP24 catalytic activity not directly demonstrated, single lab","pmids":["37187452"],"is_preprint":false}],"current_model":"USP24 is a large deubiquitinating enzyme (DUB) that stabilizes a broad set of substrates — including p53, DDB2, ULK1, CRMP2, Mcl-1, BRD-containing proteins, GSDMB, p300, β-TrCP, TRAF2, TRAF6, Runx2, Beclin1, PKA-Cα, STAT2, YAP1, DHODH, and PD-1 — by removing K48-linked (and in some contexts K63-linked) ubiquitin chains to prevent proteasomal degradation; through these substrates it regulates the DNA damage response, autophagy flux, ferroptosis, NF-κB and STAT3 signaling, mitotic fidelity, osteoblast differentiation, innate immune ISGylation (via MOV10 deISGylation), and T-cell PD-1-mediated exhaustion, while its catalytic activity (centered on Cys1695) is required for all these functions as shown by catalytic-dead mutant studies."},"narrative":{"mechanistic_narrative":"USP24 is a large cysteine-protease deubiquitinating enzyme (DUB) that acts as a master post-translational stabilizer, removing K48-linked polyubiquitin from a broad panel of substrates to spare them from proteasomal degradation, with catalytic activity centered on Cys1695 required for its functions [PMID:38491202, PMID:40238877]. Its earliest-defined roles are in the genome-stability arm: USP24 directly deubiquitinates and stabilizes p53 to enable UV-induced PUMA activation and apoptosis [PMID:25578727] and stabilizes the nucleotide excision repair factor DDB2 [PMID:23159851], while loss of USP24 destabilizes the microtubule regulator CRMP2 to cause spindle defects and aneuploidy, casting USP24 as a tumor suppressor of mitotic fidelity [PMID:33355202]. USP24 also bidirectionally tunes autophagy: it restrains flux by destabilizing ULK1 [PMID:30957634], yet in other contexts stabilizes Beclin1 to drive autophagy-dependent ferroptosis [PMID:39379617] or acts through TRAF6 to control Beclin-1 turnover [PMID:41785730]. In cancer and immune settings it sustains pro-tumorigenic signaling by stabilizing substrates including GSDMB (promoting STAT3 activation) [PMID:34326684], TRAF2 (driving AKT/NF-κB) [PMID:39127151], YAP1 [PMID:40287768], and the anti-ferroptotic enzyme DHODH [PMID:40715045], and it stabilizes PD-1 in CD8+ T cells downstream of an IL-6/STAT3 axis, linking it to T-cell exhaustion and lung tumorigenesis [PMID:40238877]. Substrate recognition is partly encoded by a C-terminal motif that engages bromodomain-containing proteins at defined ubiquitin acceptor lysines [PMID:33257797]. Beyond canonical ubiquitin, USP24 is ISG15 cross-reactive and deISGylates the RNA helicase MOV10 to negatively regulate innate IFN-β responses.","teleology":[{"year":2012,"claim":"Established USP24 as a bona fide deubiquitinase by showing it physically engages and enzymatically stabilizes a defined substrate, the NER factor DDB2.","evidence":"Yeast two-hybrid, co-IP, siRNA knockdown, and in vitro deubiquitination with purified USP24","pmids":["23159851"],"confidence":"High","gaps":["Did not define ubiquitin linkage specificity","No structural basis for substrate selection"]},{"year":2015,"claim":"Placed USP24 in the DNA-damage response by demonstrating direct deubiquitination of p53 governs UV-induced apoptosis, framing USP24 as a stress-responsive stabilizer.","evidence":"In vitro deubiquitination with purified USP24, siRNA knockdown with p53 stability/apoptosis readouts, and re-expression rescue","pmids":["25578727"],"confidence":"High","gaps":["How DNA damage triggers USP24-p53 engagement is unresolved","Linkage type not specified"]},{"year":2019,"claim":"Revealed USP24 as a regulator of autophagy and macrophage-driven inflammation, expanding it beyond DNA repair into proteostasis and immune signaling.","evidence":"siRNA knockdown in cell lines and iPSC dopaminergic neurons (ULK1/autophagy flux); knockdown in M2 macrophages/lung cancer with IL-6 and p300/β-TrCP readouts","pmids":["30957634","30266897"],"confidence":"Medium","gaps":["Direct deubiquitination of ULK1 not reconstituted in vitro","Whether p300/β-TrCP are direct substrates not shown by in vitro assay"]},{"year":2020,"claim":"Defined a substrate-recognition mechanism and a genetic tumor-suppressor role, connecting molecular substrate engagement to organismal phenotype.","evidence":"C-terminus/bromodomain interaction mapping and acceptor-lysine mutagenesis; Usp24 knockout mouse with CRMP2 rescue of spindle/segregation defects","pmids":["33257797","33355202"],"confidence":"Medium","gaps":["No structure of the USP24-BRD interface","How a single DUB selects such diverse substrates remains unexplained"]},{"year":2021,"claim":"Extended USP24 substrate stabilization to oncogenic effectors and drug-resistance machinery, linking its activity to cancer progression and chemoresistance.","evidence":"Mass spec/co-IP for GSDMB (STAT3); knockdown/overexpression with Rad51 and ABC-transporter readouts and inhibitor treatment","pmids":["34326684","33846536"],"confidence":"Medium","gaps":["Direct vs indirect effects on Rad51 and ABC transporters not separated","Linkage specificity not defined"]},{"year":2024,"claim":"Resolved ubiquitin-linkage specificity (K48) for multiple substrates and showed USP24 controls ferroptosis and survival signaling in hepatocellular carcinoma.","evidence":"Co-IP, K48-linkage-specific ubiquitination assays, genetic rescue (Beclin1, TRAF2) and xenografts","pmids":["39379617","39127151"],"confidence":"Medium","gaps":["Context-dependence of pro- vs anti-autophagy outcomes unexplained","Substrate competition/hierarchy not addressed"]},{"year":2024,"claim":"Demonstrated catalytic-dependence in vivo and integrated USP24 into autophagy-linked drug resistance and immune checkpoint control in lung cancer.","evidence":"CRISPR catalytic knockout (C1695A), USP24-i-101 inhibitor, bafilomycin epistasis, inducible EGFR-L858R mouse model","pmids":["38491202"],"confidence":"Medium","gaps":["Direct vs indirect regulation of E2F4/TRAF6 not fully separated","PD-L1 degradation mechanism not molecularly defined here"]},{"year":2024,"claim":"Identified USP24 as ISG15 cross-reactive, broadening its activity beyond ubiquitin to deISGylation of MOV10 in innate immunity.","evidence":"ABPP, in vitro deISGylation with recombinant USP24, depletion with ISG15 conjugate/interactome proteomics (preprint)","pmids":[],"confidence":"Medium","gaps":["Preprint, not peer-reviewed","Physiological balance between deubiquitination and deISGylation unclear"]},{"year":2025,"claim":"Established USP24 as a checkpoint regulator by K48-deubiquitinating PD-1 downstream of IL-6/STAT3, linking its activity to T-cell exhaustion and tumor immunity.","evidence":"Co-IP, K48-ubiquitination assay, catalytic-deficient mice, USP24-i-101 and anti-CTLA4 combination in vivo","pmids":["40238877"],"confidence":"High","gaps":["How IL-6/STAT3 transcriptionally drives USP24 mechanistically not fully mapped","Whether PD-1 stabilization is the dominant in vivo effect not isolated"]},{"year":2025,"claim":"Widened the substrate set to metabolic and growth-control effectors (DHODH, YAP1, PKA-Cα), positioning USP24 in ferroptosis resistance, proliferation, and adipocyte/metabolic signaling.","evidence":"Co-IP, deubiquitination assays, knockdown with substrate rescue, catalytic-knockout mice and RNA-seq","pmids":["40715045","40287768","40448065"],"confidence":"Medium","gaps":["Linkage specificity not defined for all substrates","Tissue-specific substrate selection rules unknown"]},{"year":2026,"claim":"Connected USP24 to osteoblast differentiation and skeletal disease via STAT2 stabilization and to cardiac autophagy via the TRAF6-Beclin1 axis.","evidence":"Co-IP, deubiquitination assays, STAT2 phenocopy, rAAV silencing in a FOP mouse model; TRAF6/Beclin-1 epistasis with autophagic flux assays","pmids":["42218160","41785730"],"confidence":"Medium","gaps":["Reconciliation of opposing autophagy effects across tissues not addressed","Direct STAT2/TRAF6 deubiquitination kinetics not reconstituted in vitro"]},{"year":null,"claim":"How a single large DUB achieves selectivity across this exceptionally broad substrate range, and what determines its context-dependent pro- versus anti-autophagy and pro- versus anti-ferroptosis outcomes, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of substrate recognition beyond the BRD-binding motif","No systematic linkage-specificity profiling","Rules governing tissue-specific substrate choice unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,1,6,10,13,14,15]},{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[0,1,19]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[2,18]}],"localization":[],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,1,6,10,13,14,15]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[2,10,12,18]},{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[0,1,8]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[13,19]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[0,5,10,14]}],"complexes":[],"partners":["TP53","DDB2","ULK1","GSDMB","CRMP2","TRAF2","TRAF6","YAP1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9UPU5","full_name":"Ubiquitin carboxyl-terminal hydrolase 24","aliases":["Deubiquitinating enzyme 24","Ubiquitin thioesterase 24","Ubiquitin-specific-processing protease 24"],"length_aa":2620,"mass_kda":294.4,"function":"Ubiquitin-specific protease that regulates cell survival in various contexts through modulating the protein stability of some of its substrates including DDB2, MCL1 or TP53. Plays a positive role on ferritinophagy where ferritin is degraded in lysosomes and releases free iron","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/Q9UPU5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/USP24","classification":"Not Classified","n_dependent_lines":53,"n_total_lines":1208,"dependency_fraction":0.043874172185430466},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/USP24","total_profiled":1310},"omim":[{"mim_id":"610569","title":"UBIQUITIN-SPECIFIC PROTEASE 24; USP24","url":"https://www.omim.org/entry/610569"},{"mim_id":"606852","title":"PARKINSON DISEASE 10; PARK10","url":"https://www.omim.org/entry/606852"},{"mim_id":"603199","title":"PALS1-ASSOCIATED TIGHT JUNCTION PROTEIN; PATJ","url":"https://www.omim.org/entry/603199"}],"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/USP24"},"hgnc":{"alias_symbol":["KIAA1057"],"prev_symbol":[]},"alphafold":{"accession":"Q9UPU5","domains":[{"cath_id":"-","chopping":"122-282","consensus_level":"medium","plddt":85.5527,"start":122,"end":282},{"cath_id":"3.90.70.10","chopping":"1691-1925_1951-2020_2035-2042","consensus_level":"medium","plddt":84.3469,"start":1691,"end":2042},{"cath_id":"-","chopping":"2463-2539_2547-2574","consensus_level":"medium","plddt":82.0603,"start":2463,"end":2574},{"cath_id":"1.10.8","chopping":"2-43","consensus_level":"high","plddt":65.98,"start":2,"end":43}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9UPU5","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9UPU5-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9UPU5-F1-predicted_aligned_error_v6.png","plddt_mean":75.12},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=USP24","jax_strain_url":"https://www.jax.org/strain/search?query=USP24"},"sequence":{"accession":"Q9UPU5","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9UPU5.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9UPU5/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9UPU5"}},"corpus_meta":[{"pmid":"34326684","id":"PMC_34326684","title":"USP24-GSDMB complex promotes bladder cancer proliferation via activation of the STAT3 pathway.","date":"2021","source":"International journal of biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/34326684","citation_count":101,"is_preprint":false},{"pmid":"30266897","id":"PMC_30266897","title":"USP24 induces IL-6 in tumor-associated microenvironment by stabilizing p300 and β-TrCP and promotes cancer malignancy.","date":"2018","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/30266897","citation_count":98,"is_preprint":false},{"pmid":"16917932","id":"PMC_16917932","title":"Genetic evidence for ubiquitin-specific proteases USP24 and USP40 as candidate genes for late-onset Parkinson disease.","date":"2006","source":"Human mutation","url":"https://pubmed.ncbi.nlm.nih.gov/16917932","citation_count":53,"is_preprint":false},{"pmid":"38056575","id":"PMC_38056575","title":"Upregulation of NF-κB by USP24 aggravates ferroptosis in diabetic cardiomyopathy.","date":"2023","source":"Free radical biology & medicine","url":"https://pubmed.ncbi.nlm.nih.gov/38056575","citation_count":51,"is_preprint":false},{"pmid":"25578727","id":"PMC_25578727","title":"The deubiquitinating enzyme USP24 is a regulator of the UV damage response.","date":"2015","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/25578727","citation_count":49,"is_preprint":false},{"pmid":"30957634","id":"PMC_30957634","title":"The PARK10 gene USP24 is a negative regulator of autophagy and ULK1 protein stability.","date":"2019","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/30957634","citation_count":43,"is_preprint":false},{"pmid":"23159851","id":"PMC_23159851","title":"The deubiquitinating protein USP24 interacts with DDB2 and regulates DDB2 stability.","date":"2012","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/23159851","citation_count":41,"is_preprint":false},{"pmid":"37187452","id":"PMC_37187452","title":"MiR-21-5p promotes sorafenib resistance and hepatocellular carcinoma progression by regulating SIRT7 ubiquitination through USP24.","date":"2023","source":"Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/37187452","citation_count":29,"is_preprint":false},{"pmid":"30911287","id":"PMC_30911287","title":"WP1130 reveals USP24 as a novel target in T-cell acute lymphoblastic leukemia.","date":"2019","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/30911287","citation_count":29,"is_preprint":false},{"pmid":"38491202","id":"PMC_38491202","title":"USP24-i-101 targeting of USP24 activates autophagy to inhibit drug resistance acquired during cancer therapy.","date":"2024","source":"Cell death and 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Ubiquitin Hydrolase, Novel Tumor Suppressor, and Chromosome Instability Gene Deleted in Neuroblastoma.","date":"2020","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/33355202","citation_count":19,"is_preprint":false},{"pmid":"37415159","id":"PMC_37415159","title":"USP24-dependent stabilization of Runx2 recruits a p300/NCOA3 complex to transactivate ADAMTS genes and promote degeneration of intervertebral disc in chronic inflammation mice.","date":"2023","source":"Biology direct","url":"https://pubmed.ncbi.nlm.nih.gov/37415159","citation_count":18,"is_preprint":false},{"pmid":"33257797","id":"PMC_33257797","title":"USP24 stabilizes bromodomain containing proteins to promote lung cancer malignancy.","date":"2020","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/33257797","citation_count":17,"is_preprint":false},{"pmid":"38140768","id":"PMC_38140768","title":"NCI677397 targeting USP24-mediated induction of lipid peroxidation induces ferroptosis in drug-resistant cancer cells.","date":"2023","source":"Molecular oncology","url":"https://pubmed.ncbi.nlm.nih.gov/38140768","citation_count":15,"is_preprint":false},{"pmid":"40238877","id":"PMC_40238877","title":"Deubiquitinase USP24 activated by IL-6/STAT3 enhances PD-1 protein stability and suppresses T cell antitumor response.","date":"2025","source":"Science advances","url":"https://pubmed.ncbi.nlm.nih.gov/40238877","citation_count":13,"is_preprint":false},{"pmid":"22923019","id":"PMC_22923019","title":"Association analysis of single-nucleotide polymorphisms of USP24 and USP40 with Parkinson's disease in the Han Chinese population.","date":"2012","source":"European neurology","url":"https://pubmed.ncbi.nlm.nih.gov/22923019","citation_count":12,"is_preprint":false},{"pmid":"40715045","id":"PMC_40715045","title":"The deubiquitinase USP24 suppresses ferroptosis in triple-negative breast cancer by stabilizing DHODH protein.","date":"2025","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/40715045","citation_count":10,"is_preprint":false},{"pmid":"39127151","id":"PMC_39127151","title":"USP24 promotes hepatocellular carcinoma tumorigenesis through deubiquitinating and stabilizing TRAF2.","date":"2024","source":"Biochemical pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/39127151","citation_count":10,"is_preprint":false},{"pmid":"40448065","id":"PMC_40448065","title":"USP24 upregulation stabilizes PKA-Cα to promote lipogenesis, inflammation, and fibrosis during MASH progression.","date":"2025","source":"Journal of biomedical science","url":"https://pubmed.ncbi.nlm.nih.gov/40448065","citation_count":4,"is_preprint":false},{"pmid":"29738159","id":"PMC_29738159","title":"[Expression characteristics of the USP24 gene in the mouse testis during spermatogenesis].","date":"2017","source":"Zhonghua nan ke xue = National journal of andrology","url":"https://pubmed.ncbi.nlm.nih.gov/29738159","citation_count":3,"is_preprint":false},{"pmid":"40287768","id":"PMC_40287768","title":"USP24 promotes hepatocellular carcinoma progression by deubiquitinating and stabilizing YAP1.","date":"2025","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/40287768","citation_count":2,"is_preprint":false},{"pmid":"40233501","id":"PMC_40233501","title":"The combination of USP24-i-101-Astemizole sensitizes the cytotoxicity of Taxol and Gefitinib in drug-resistant lung cancer.","date":"2025","source":"Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie","url":"https://pubmed.ncbi.nlm.nih.gov/40233501","citation_count":0,"is_preprint":false},{"pmid":"42218160","id":"PMC_42218160","title":"USP24-dependent STAT2 stabilization mediates physiologic and pathologic bone formation.","date":"2026","source":"Cell death & 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Lymphoma","date":"2025-09-20","source":"bioRxiv","url":"https://doi.org/10.1101/2025.09.18.675797","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.09.06.611391","title":"USP24 is an ISG15 cross-reactive deubiquitinase that mediates IFN-I production by de-ISGylating the RNA helicase MOV10","date":"2024-09-06","source":"bioRxiv","url":"https://doi.org/10.1101/2024.09.06.611391","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":16784,"output_tokens":5333,"usd":0.065173,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":13732,"output_tokens":3943,"usd":0.083617,"stage2_stop_reason":"end_turn"},"total_usd":0.14879,"stage1_batch_id":"msgbatch_01UT1AahFPCAyn2uVweS7A9z","stage2_batch_id":"msgbatch_01HuKWtKNT4Pqj1bc8eEP5RV","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2015,\n      \"finding\": \"USP24 directly deubiquitinates p53, stabilizing it in response to UV-induced DNA damage; purified USP24 protein cleaves ubiquitinated p53 in vitro, and USP24 depletion destabilizes p53, impairs PUMA activation, and renders cells resistant to apoptosis after UV irradiation.\",\n      \"method\": \"In vitro deubiquitination assay with purified USP24; siRNA knockdown with p53 stability and apoptosis readouts; forced re-expression rescue experiment\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with purified protein plus cellular knockdown/rescue with multiple orthogonal readouts in a single focused study\",\n      \"pmids\": [\"25578727\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"USP24 interacts with DDB2 (a nucleotide excision repair factor) and stabilizes it by removing ubiquitin; USP24 knockdown decreases steady-state DDB2 levels, and purified USP24 cleaves ubiquitinated DDB2 in vitro.\",\n      \"method\": \"Yeast two-hybrid screen; co-immunoprecipitation; siRNA knockdown; in vitro deubiquitination assay\",\n      \"journal\": \"Cell cycle (Georgetown, Tex.)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution plus reciprocal interaction confirmation and knockdown, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"23159851\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"USP24 negatively regulates autophagy by controlling ubiquitination and stability of ULK1; USP24 knockdown in cell lines and iPSC-derived dopaminergic neurons elevates ULK1 protein levels and increases autophagy flux in an MTORC1-independent, class III PtdIns3K-dependent manner.\",\n      \"method\": \"siRNA knockdown in cell lines and iPSC-derived dopaminergic neurons; autophagy flux assays; MTORC1 and PtdIns3K inhibitor epistasis\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean knockdown with defined cellular phenotype and pathway placement via pharmacological epistasis, single lab\",\n      \"pmids\": [\"30957634\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"USP24 stabilizes p300 and β-TrCP proteins in M2 macrophages and lung cancer cells, leading to increased histone-3 acetylation and NF-κB levels while decreasing DNMT1 and IκB, thereby increasing IL-6 transcription.\",\n      \"method\": \"siRNA knockdown; western blot for substrate levels; conditioned medium transfer assays; IL-6 rescue experiments\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — knockdown with defined molecular mechanism across two cell types, multiple downstream readouts, single lab\",\n      \"pmids\": [\"30266897\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"USP24 interacts with GSDMB and prevents its proteasomal degradation (deubiquitination-mediated stabilization), and the USP24/GSDMB complex promotes STAT3 phosphorylation and activation in bladder cancer cells.\",\n      \"method\": \"Mass spectrometry; co-immunoprecipitation; western blot; USP24 inhibitor treatment\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-IP confirmed interaction and stabilization across multiple experiments, single lab\",\n      \"pmids\": [\"34326684\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"USP24 stabilizes Mcl-1; WP1130 inhibits USP24 catalytic activity (molecular docking and cellular thermal shift assay), leading to Mcl-1 destabilization and apoptosis via mitochondrial transmembrane potential collapse in T-ALL cells.\",\n      \"method\": \"Molecular docking; cellular thermal shift assay (CETSA); siRNA knockdown; Mcl-1 protein level measurement; mitochondrial transmembrane potential assay\",\n      \"journal\": \"Cancer cell international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CETSA confirms direct drug-target engagement, knockdown phenocopies drug effect, single lab\",\n      \"pmids\": [\"30911287\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"USP24 interacts with the bromodomain (BRD) via its C-terminus and stabilizes most BRD-containing proteins by removing ubiquitin from Lys391/Lys400 of the BRD; loss of the USP24-BRD interaction motif abolishes this stabilization and USP24's effects on cancer progression.\",\n      \"method\": \"Co-immunoprecipitation; western blot for BRD protein stability; mutagenesis of ubiquitin acceptor sites (Lys391/Lys400); deletion mutant analysis\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — site-directed mutagenesis of substrate ubiquitination sites plus interaction mapping, single lab\",\n      \"pmids\": [\"33257797\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"USP24 interacts with and stabilizes CRMP2; deletion of Usp24 in mice leads to CRMP2 degradation, causing spindle defects, chromosome missegregation, and aneuploidy that are rescued by CRMP2 restoration, identifying USP24 as a tumor suppressor maintaining mitotic accuracy.\",\n      \"method\": \"Usp24 knockout mouse model; genetic rescue with CRMP2 restoration; chromosome segregation and spindle defect assays\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic rescue experiment in murine knockout model with defined cellular phenotype, single lab\",\n      \"pmids\": [\"33355202\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"USP24 represses DNA-damage repair by decreasing Rad51 expression (promoting genomic instability) and increases ABC transporter levels (P-gp, ABCG2, ezrin) to enhance Taxol efflux from cancer cells, thereby promoting drug resistance.\",\n      \"method\": \"USP24 knockdown/overexpression; western blot for Rad51 and ABC transporter levels; drug efflux assays; NCI677397 inhibitor treatment\",\n      \"journal\": \"Cell death and differentiation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — knockdown with defined molecular readouts and pharmacological validation, single lab\",\n      \"pmids\": [\"33846536\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"USP24 is recruited to phospho-Runx2 (pRunx2, phosphorylated at Ser28 by p38 kinase) and stabilizes pRunx2 by deubiquitination, allowing pRunx2 to assemble a NCOA3-p300-pRunx2 complex that transactivates ADAMTS genes and promotes ECM degradation in intervertebral disc degeneration.\",\n      \"method\": \"Co-immunoprecipitation; mass spectrometry; western blot; RT-qPCR; p38 inhibitor epistasis; LPS-induced chronic inflammation mouse model\",\n      \"journal\": \"Biology direct\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP/MS confirmed complex, pharmacological epistasis defined pathway, single lab\",\n      \"pmids\": [\"37415159\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"USP24 reduces K48-linked ubiquitination of Beclin1, thereby stabilizing it and promoting autophagy-dependent ferroptosis in hepatocellular carcinoma; effects of USP24 overexpression on HCC proliferation are partially reversed by Beclin1 silencing.\",\n      \"method\": \"Co-immunoprecipitation confirming endogenous USP24-Beclin1 interaction; ubiquitination assay (K48-linkage specific); genetic rescue with Beclin1 silencing; in vivo xenograft\",\n      \"journal\": \"Communications biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus K48-specific ubiquitination assay and genetic epistasis, single lab\",\n      \"pmids\": [\"39379617\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"USP24 binds to and deubiquitinates TRAF2, preventing its degradation; stabilized TRAF2 activates the AKT/NF-κB signaling pathway to promote HCC cell survival.\",\n      \"method\": \"Co-immunoprecipitation; western blot for TRAF2 stability after USP24 manipulation; AKT/NF-κB pathway readouts\",\n      \"journal\": \"Biochemical pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-IP and substrate stabilization confirmed across multiple experiments, single lab\",\n      \"pmids\": [\"39127151\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"USP24 activates autophagy in both interphase and mitotic periods of lung cancer cells by inhibiting E2F4 and TRAF6, respectively; the catalytic mutant USP24C1695A abolishes this effect, and autophagy induction is required for USP24 inhibitor-mediated suppression of drug resistance and maintenance of genomic integrity and PD-L1 degradation.\",\n      \"method\": \"CRISPR-mediated catalytic knockout (USP24C1695A); USP24 inhibitor USP24-i-101; bafilomycin-A1 autophagy inhibition rescue; doxycycline-inducible EGFRL858R lung cancer mouse model\",\n      \"journal\": \"Cell death and differentiation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — catalytic mutant confirms enzyme-dependent mechanism, in vivo mouse model, autophagy epistasis, single lab\",\n      \"pmids\": [\"38491202\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"USP24 removes K48-linked polyubiquitin from PD-1, stabilizing PD-1 protein in CD8+ T cells; IL-6 transcriptionally activates USP24 expression, linking the IL-6/STAT3 axis to PD-1 stabilization; USP24 catalytic-deficient (Usp24 catalytic mutant) mice show reduced PD-1 levels and attenuated lung tumorigenesis.\",\n      \"method\": \"Co-immunoprecipitation; K48-ubiquitination assay; catalytic-deficient mouse model; USP24-i-101 inhibitor treatment; anti-CTLA4 combination in vivo\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — specific ubiquitin linkage identified, in vivo genetic catalytic-mutant model, pharmacological validation, multiple orthogonal methods in one study\",\n      \"pmids\": [\"40238877\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"USP24 directly interacts with and deubiquitinates DHODH, stabilizing it to maintain coenzyme Q reduction and protect TNBC cells from lipid peroxidation-driven ferroptosis; pharmacological USP24 inhibition synergizes with ferroptosis inducers via a DHODH-dependent pathway.\",\n      \"method\": \"Co-immunoprecipitation; deubiquitination assay; siRNA knockdown; in vitro and in vivo ferroptosis assays with DHODH rescue\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus DHODH-dependent genetic rescue and pharmacological validation, single lab\",\n      \"pmids\": [\"40715045\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"USP24 stabilizes YAP1 by directly interacting with and deubiquitinating it; USP24 depletion suppresses HCC cell proliferation and tumor growth, and these effects are rescued by YAP1 restoration.\",\n      \"method\": \"Co-immunoprecipitation; deubiquitination assay; siRNA knockdown; YAP1 rescue experiment; xenograft mouse model\",\n      \"journal\": \"Cancer cell international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, deubiquitination assay, and genetic rescue in vivo, single lab\",\n      \"pmids\": [\"40287768\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"USP24 interacts with and stabilizes PKA-Cα by deubiquitination; USP24 expression is upregulated by PKA activation during adipocyte differentiation, creating a positive feedback loop that increases CREB phosphorylation and lipogenic gene expression; free fatty acids increase USP24 to activate NF-κB and TGFβ pathways driving inflammation and fibrosis.\",\n      \"method\": \"Co-immunoprecipitation; CRISPR/Cas9 catalytic knockout (USP24C1695A) mice; USP24-i-101 inhibitor; western blot; RNA-sequencing\",\n      \"journal\": \"Journal of biomedical science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus catalytic knockout mouse model and transcriptomic data, single lab\",\n      \"pmids\": [\"40448065\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"USP24 stabilizes STAT2 through deubiquitination in osteoblasts; loss of USP24 reduces osteoblast differentiation and bone formation, phenocopying STAT2 deficiency; rAAV-mediated Usp24 silencing in a FOP mouse model diminishes heterotopic ossification and reduces STAT2 protein levels.\",\n      \"method\": \"Co-immunoprecipitation; deubiquitination assay; USP24 knockout/knockdown; STAT2 knockdown phenocopy; rAAV-mediated silencing in FOP mouse model\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic interaction confirmed by phenocopy and in vivo model, single lab\",\n      \"pmids\": [\"42218160\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"USP24 deubiquitinates TRAF6, stabilizing it; stabilized TRAF6 then ubiquitinates Beclin-1 to promote its degradation, thereby inhibiting autophagy in diabetic myocardial disorder; TRAF6 overexpression reverses the autophagy rescue caused by USP24 knockdown.\",\n      \"method\": \"Co-immunoprecipitation; western blot for TRAF6 and Beclin-1 levels; lentiviral knockdown/overexpression; autophagic flux assays (mRFP-GFP-LC3, TEM); genetic epistasis (TRAF6 OE reverses USP24 KD effect)\",\n      \"journal\": \"Life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, substrate ubiquitination, and genetic epistasis, single lab\",\n      \"pmids\": [\"41785730\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"USP24 is an ISG15 cross-reactive deubiquitylase; recombinant USP24 processes pro-ISG15 and cleaves ISG15-linked synthetic substrates in vitro; USP24 depletion increases ISG15 conjugates upon IFN-β stimulation; USP24 specifically deISGylates the RNA helicase MOV10, and ISGylated MOV10 enhances IFN-β production, while USP24-mediated deISGylation of MOV10 negatively regulates the innate immune response.\",\n      \"method\": \"Activity-based protein profiling (ABPP); in vitro deISGylation assay with recombinant USP24 and synthetic ISG15-substrates; USP24 depletion; total proteome, GG-peptidome, and ISG15 interactome proteomics\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — in vitro reconstitution with recombinant protein plus proteomics, but preprint not yet peer-reviewed, single study\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"USP24 promotes NF-κB accumulation in diabetic cardiomyopathy; USP24 knockdown reduces NF-κB levels, whereas NF-κB knockdown does not reduce USP24 expression, establishing USP24 as upstream of NF-κB; elevated NF-κB promotes FACL4 expression and ferroptosis while reducing SLC7A11 and FTH1.\",\n      \"method\": \"siRNA knockdown in H9c2 cells; PMA-mediated NF-κB re-activation rescue; western blot; in vivo HFD/STZ mouse model\",\n      \"journal\": \"Free radical biology & medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — directional epistasis established by unidirectional knockdown and pharmacological rescue, single lab\",\n      \"pmids\": [\"38056575\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"USP24 interacts with SIRT7 and mediates its deubiquitination; inhibition of miR-21-5p downregulates the ubiquitination level of SIRT7 via USP24, affecting autophagy levels in hepatocellular carcinoma.\",\n      \"method\": \"Co-immunoprecipitation; western blot for SIRT7 ubiquitination\",\n      \"journal\": \"Life sciences\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single co-IP experiment, mechanistic link to USP24 catalytic activity not directly demonstrated, single lab\",\n      \"pmids\": [\"37187452\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"USP24 is a large deubiquitinating enzyme (DUB) that stabilizes a broad set of substrates — including p53, DDB2, ULK1, CRMP2, Mcl-1, BRD-containing proteins, GSDMB, p300, β-TrCP, TRAF2, TRAF6, Runx2, Beclin1, PKA-Cα, STAT2, YAP1, DHODH, and PD-1 — by removing K48-linked (and in some contexts K63-linked) ubiquitin chains to prevent proteasomal degradation; through these substrates it regulates the DNA damage response, autophagy flux, ferroptosis, NF-κB and STAT3 signaling, mitotic fidelity, osteoblast differentiation, innate immune ISGylation (via MOV10 deISGylation), and T-cell PD-1-mediated exhaustion, while its catalytic activity (centered on Cys1695) is required for all these functions as shown by catalytic-dead mutant studies.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"USP24 is a large cysteine-protease deubiquitinating enzyme (DUB) that acts as a master post-translational stabilizer, removing K48-linked polyubiquitin from a broad panel of substrates to spare them from proteasomal degradation, with catalytic activity centered on Cys1695 required for its functions [#12, #13]. Its earliest-defined roles are in the genome-stability arm: USP24 directly deubiquitinates and stabilizes p53 to enable UV-induced PUMA activation and apoptosis [#0] and stabilizes the nucleotide excision repair factor DDB2 [#1], while loss of USP24 destabilizes the microtubule regulator CRMP2 to cause spindle defects and aneuploidy, casting USP24 as a tumor suppressor of mitotic fidelity [#7]. USP24 also bidirectionally tunes autophagy: it restrains flux by destabilizing ULK1 [#2], yet in other contexts stabilizes Beclin1 to drive autophagy-dependent ferroptosis [#10] or acts through TRAF6 to control Beclin-1 turnover [#18]. In cancer and immune settings it sustains pro-tumorigenic signaling by stabilizing substrates including GSDMB (promoting STAT3 activation) [#4], TRAF2 (driving AKT/NF-\\u03baB) [#11], YAP1 [#15], and the anti-ferroptotic enzyme DHODH [#14], and it stabilizes PD-1 in CD8+ T cells downstream of an IL-6/STAT3 axis, linking it to T-cell exhaustion and lung tumorigenesis [#13]. Substrate recognition is partly encoded by a C-terminal motif that engages bromodomain-containing proteins at defined ubiquitin acceptor lysines [#6]. Beyond canonical ubiquitin, USP24 is ISG15 cross-reactive and deISGylates the RNA helicase MOV10 to negatively regulate innate IFN-\\u03b2 responses [#19].\",\n  \"teleology\": [\n    {\n      \"year\": 2012,\n      \"claim\": \"Established USP24 as a bona fide deubiquitinase by showing it physically engages and enzymatically stabilizes a defined substrate, the NER factor DDB2.\",\n      \"evidence\": \"Yeast two-hybrid, co-IP, siRNA knockdown, and in vitro deubiquitination with purified USP24\",\n      \"pmids\": [\"23159851\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not define ubiquitin linkage specificity\", \"No structural basis for substrate selection\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Placed USP24 in the DNA-damage response by demonstrating direct deubiquitination of p53 governs UV-induced apoptosis, framing USP24 as a stress-responsive stabilizer.\",\n      \"evidence\": \"In vitro deubiquitination with purified USP24, siRNA knockdown with p53 stability/apoptosis readouts, and re-expression rescue\",\n      \"pmids\": [\"25578727\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How DNA damage triggers USP24-p53 engagement is unresolved\", \"Linkage type not specified\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Revealed USP24 as a regulator of autophagy and macrophage-driven inflammation, expanding it beyond DNA repair into proteostasis and immune signaling.\",\n      \"evidence\": \"siRNA knockdown in cell lines and iPSC dopaminergic neurons (ULK1/autophagy flux); knockdown in M2 macrophages/lung cancer with IL-6 and p300/\\u03b2-TrCP readouts\",\n      \"pmids\": [\"30957634\", \"30266897\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct deubiquitination of ULK1 not reconstituted in vitro\", \"Whether p300/\\u03b2-TrCP are direct substrates not shown by in vitro assay\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Defined a substrate-recognition mechanism and a genetic tumor-suppressor role, connecting molecular substrate engagement to organismal phenotype.\",\n      \"evidence\": \"C-terminus/bromodomain interaction mapping and acceptor-lysine mutagenesis; Usp24 knockout mouse with CRMP2 rescue of spindle/segregation defects\",\n      \"pmids\": [\"33257797\", \"33355202\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structure of the USP24-BRD interface\", \"How a single DUB selects such diverse substrates remains unexplained\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Extended USP24 substrate stabilization to oncogenic effectors and drug-resistance machinery, linking its activity to cancer progression and chemoresistance.\",\n      \"evidence\": \"Mass spec/co-IP for GSDMB (STAT3); knockdown/overexpression with Rad51 and ABC-transporter readouts and inhibitor treatment\",\n      \"pmids\": [\"34326684\", \"33846536\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect effects on Rad51 and ABC transporters not separated\", \"Linkage specificity not defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Resolved ubiquitin-linkage specificity (K48) for multiple substrates and showed USP24 controls ferroptosis and survival signaling in hepatocellular carcinoma.\",\n      \"evidence\": \"Co-IP, K48-linkage-specific ubiquitination assays, genetic rescue (Beclin1, TRAF2) and xenografts\",\n      \"pmids\": [\"39379617\", \"39127151\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Context-dependence of pro- vs anti-autophagy outcomes unexplained\", \"Substrate competition/hierarchy not addressed\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstrated catalytic-dependence in vivo and integrated USP24 into autophagy-linked drug resistance and immune checkpoint control in lung cancer.\",\n      \"evidence\": \"CRISPR catalytic knockout (C1695A), USP24-i-101 inhibitor, bafilomycin epistasis, inducible EGFR-L858R mouse model\",\n      \"pmids\": [\"38491202\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect regulation of E2F4/TRAF6 not fully separated\", \"PD-L1 degradation mechanism not molecularly defined here\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified USP24 as ISG15 cross-reactive, broadening its activity beyond ubiquitin to deISGylation of MOV10 in innate immunity.\",\n      \"evidence\": \"ABPP, in vitro deISGylation with recombinant USP24, depletion with ISG15 conjugate/interactome proteomics (preprint)\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint, not peer-reviewed\", \"Physiological balance between deubiquitination and deISGylation unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Established USP24 as a checkpoint regulator by K48-deubiquitinating PD-1 downstream of IL-6/STAT3, linking its activity to T-cell exhaustion and tumor immunity.\",\n      \"evidence\": \"Co-IP, K48-ubiquitination assay, catalytic-deficient mice, USP24-i-101 and anti-CTLA4 combination in vivo\",\n      \"pmids\": [\"40238877\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How IL-6/STAT3 transcriptionally drives USP24 mechanistically not fully mapped\", \"Whether PD-1 stabilization is the dominant in vivo effect not isolated\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Widened the substrate set to metabolic and growth-control effectors (DHODH, YAP1, PKA-C\\u03b1), positioning USP24 in ferroptosis resistance, proliferation, and adipocyte/metabolic signaling.\",\n      \"evidence\": \"Co-IP, deubiquitination assays, knockdown with substrate rescue, catalytic-knockout mice and RNA-seq\",\n      \"pmids\": [\"40715045\", \"40287768\", \"40448065\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Linkage specificity not defined for all substrates\", \"Tissue-specific substrate selection rules unknown\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Connected USP24 to osteoblast differentiation and skeletal disease via STAT2 stabilization and to cardiac autophagy via the TRAF6-Beclin1 axis.\",\n      \"evidence\": \"Co-IP, deubiquitination assays, STAT2 phenocopy, rAAV silencing in a FOP mouse model; TRAF6/Beclin-1 epistasis with autophagic flux assays\",\n      \"pmids\": [\"42218160\", \"41785730\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Reconciliation of opposing autophagy effects across tissues not addressed\", \"Direct STAT2/TRAF6 deubiquitination kinetics not reconstituted in vitro\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How a single large DUB achieves selectivity across this exceptionally broad substrate range, and what determines its context-dependent pro- versus anti-autophagy and pro- versus anti-ferroptosis outcomes, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of substrate recognition beyond the BRD-binding motif\", \"No systematic linkage-specificity profiling\", \"Rules governing tissue-specific substrate choice unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 1, 6, 10, 13, 14, 15]},\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [0, 1, 19]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [2, 18]}\n    ],\n    \"localization\": [],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 1, 6, 10, 13, 14, 15]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [2, 10, 12, 18]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [0, 1, 8]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [13, 19]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [0, 5, 10, 14]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"TP53\", \"DDB2\", \"ULK1\", \"GSDMB\", \"CRMP2\", \"TRAF2\", \"TRAF6\", \"YAP1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}