{"gene":"PSMD14","run_date":"2026-06-10T06:43:36","timeline":{"discoveries":[{"year":2002,"finding":"Rpn11 is a metalloprotease deubiquitinase (DUB) residing in the proteasome lid subcomplex. Its JAMM motif (EX(n)HXHX(10)D) constitutes the active site; mutation of the predicted catalytic histidines to alanine (rpn11AXA) was lethal in yeast and caused mutant proteasomes—which assembled normally—to fail to deubiquitinate or degrade ubiquitinated Sic1 in vitro, demonstrating that Rpn11-mediated deubiquitination is coupled to substrate degradation.","method":"Active-site mutagenesis (rpn11AXA), yeast genetics (lethality), in vitro reconstituted degradation assay with purified mutant proteasomes","journal":"Science","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with mutagenesis, functional coupling demonstrated, independently replicated across multiple subsequent studies","pmids":["12183636"],"is_preprint":false},{"year":2002,"finding":"The MPN+ motif (five conserved polar residues resembling metalloprotease active-site residues) is essential for Rpn11 function. Single amino acid substitutions in MPN+ residues cause slow growth, temperature sensitivity, and proteasome-dependent proteolysis defects in yeast, while a conserved Cys outside MPN+ is not essential.","method":"Site-directed mutagenesis of MPN+ motif residues, yeast phenotypic analysis, proteasome proteolysis assays","journal":"BMC Biochemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — mutagenesis with multiple mutant alleles, consistent phenotypes, replicated findings","pmids":["12370088"],"is_preprint":false},{"year":2003,"finding":"Rpn11 and Ubp6 serve complementary roles in proteasomal deubiquitination. Proteasomes purified from rpn11 catalytic-motif mutants or ubp6 null strains both show slower deubiquitination rates; the double mutant is synthetically lethal. Rpn11-containing proteasomes show sensitivity to metal chelators consistent with Rpn11 being a metalloprotein. Degradation requires the intact lid (lidless proteasomes deubiquitinate but do not degrade ubiquitinated protein).","method":"Purification of proteasomes from yeast mutants, in vitro deubiquitination assays, metal chelator treatment, synthetic lethality genetics","journal":"Journal of Biological Chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal biochemical approaches, genetic epistasis, replicated in subsequent studies","pmids":["14581483"],"is_preprint":false},{"year":2004,"finding":"Rpn11 has two separable functional domains: its N-terminal MPN+/JAMM catalytic domain mediates proteasome-associated deubiquitination required for proteolysis, while a distinct C-terminal domain is required for mitochondrial morphology maintenance and is independent of catalytic activity. Overexpression of wild-type Rpn8 rescues cell cycle but not mitochondrial defects of rpn11 C-terminal mutant (mpr1-1). RPN8-RPN11 chimera analysis confirmed the C-terminal region of Rpn11 is necessary and sufficient to rescue mitochondrial phenotypes.","method":"Yeast genetics, RPN8-RPN11 chimera expression, intragenic complementation, mitochondrial morphology microscopy, cell cycle analysis","journal":"Biochemical Journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple chimeric constructs, complementation assays, functional dissection of two independent domains replicated across multiple studies","pmids":["15018611"],"is_preprint":false},{"year":2006,"finding":"POH1/PSMD14 deubiquitinates c-Jun in mammalian cells. Ectopic POH1 expression in HEK293 cells decreased c-Jun ubiquitination, stabilized c-Jun protein, redistributed it to the nucleus, and increased AP1-mediated gene expression. Mutation of Cys-120 in the MPN+ motif reduced these effects. The stabilization appeared selective for c-Jun among tested proteasomal substrates.","method":"Ectopic overexpression, ubiquitination assays, reporter assays for AP1 transcription, active-site mutagenesis (C120 mutation)","journal":"Journal of Biological Chemistry","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single lab, Co-IP/ubiquitination assay plus functional reporter, selectivity claim based on limited substrate panel","pmids":["16569633"],"is_preprint":false},{"year":2007,"finding":"The JAMM zinc metalloproteinase motif of human POH1/PSMD14 is essential for cell viability and 26S proteasome function. RNAi knockdown of endogenous POH1 reduced viability and elevated polyubiquitinated proteins; rescue with wild-type POH1 restored viability, but a JAMM active-site double histidine-to-alanine mutant failed to rescue, confirming catalytic activity is required.","method":"siRNA knockdown, RNAi complementation with wild-type vs. JAMM-mutant POH1, cell viability assays, polyubiquitin accumulation analysis in HeLa cells","journal":"Molecular Cancer Therapeutics","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — RNAi complementation with active-site mutant, functional rescue assay, single lab but orthogonal methods","pmids":["17237285"],"is_preprint":false},{"year":2009,"finding":"POH1/PSMD14 within the PA700/19S proteasome is a K63-specific deubiquitinase. Biochemical fractionation of HeLa cell extracts through seven chromatographic steps co-purified the K63-specific DUB activity with the 19S proteasome. This activity was intrinsic to PA700 and was insensitive to N-ethylmaleimide and ubiquitin aldehyde (ruling out cysteine-based DUBs), consistent with JAMM metalloprotease activity. None of the complexes cleaved K6, K11, K29, K48, or alpha-linked chains.","method":"Multi-step chromatographic co-fractionation, linkage-specific deubiquitination assays, inhibitor profiling (NEM, ubiquitin aldehyde), gel filtration","journal":"EMBO Journal","confidence":"High","confidence_rationale":"Tier 1 / Moderate — seven-step biochemical co-purification plus multiple linkage-specificity assays, single lab but rigorous biochemistry","pmids":["19214193"],"is_preprint":false},{"year":2009,"finding":"POH1/PSMD14 knockdown causes apparent loss of ErbB2 protein in HeLa cells, explained at least partially by accumulation of higher-molecular-weight ubiquitinated forms of ErbB2 rather than increased degradation rate. POH1 appears to deubiquitinate ErbB2 in a manner not necessarily coupled to proteasomal degradation. Cell-surface ErbB2 levels were only mildly affected.","method":"siRNA library screen for DUBs, Western blot, flow cytometry for surface ErbB2, comparison with proteasome inhibitor epoxomicin","journal":"PLoS ONE","confidence":"Medium","confidence_rationale":"Tier 3 / Weak — single Co-IP/knockdown approach, single lab, partial mechanistic follow-up","pmids":["19436748"],"is_preprint":false},{"year":2009,"finding":"PSMD14 knockdown in carcinoma cell lines causes G0/G1 cell cycle arrest and cellular senescence, associated with downregulation of cyclin B1-CDK1-CDC25C and cyclin D1, upregulation of p21 and p27, and markedly reduced retinoblastoma protein phosphorylation. These effects are distinct from those caused by knockdown of PSMB5 (20S subunit), indicating that the 19S and 20S proteasome subunits have distinct biological functions.","method":"siRNA knockdown, flow cytometry (cell cycle), Western blot for cell cycle regulators, comparative siRNA of PSMB5","journal":"Experimental Cell Research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KD with specific phenotypic readouts, comparison with 20S component, single lab","pmids":["19732767"],"is_preprint":false},{"year":2012,"finding":"POH1/PSMD14 processes K63-linked polyubiquitin chains generated at DNA double-strand break (DSB) sites, thereby limiting 53BP1 accumulation (via antagonism of RNF8/RNF168-mediated K63-Ub) and promoting JMJD2A chromatin retention. POH1 also promotes RAD51 loading in homologous recombination independently of 53BP1. POH1-deficient cells show increased sensitivity to DNA damaging agents.","method":"siRNA knockdown, immunofluorescence for DSB foci (53BP1, RAD51, γH2AX), K63-Ub chromatin analysis, cell survival assays","journal":"EMBO Journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods, multiple pathway endpoints, functionally linked to DNA repair mechanism","pmids":["22909820"],"is_preprint":false},{"year":2013,"finding":"POH1 relieves the barrier posed by RAP80 to DNA end resection in G2 phase. POH1 depletion enlarges 53BP1 and ubiquitin chain foci and prevents formation of an RPA-positive resection core. Co-depletion of POH1 with RAP80, BRCC36, or ABRAXAS restores the resection-competent core, suggesting POH1 removes ubiquitin chains in the IRIF core that RAP80 reads, enabling transition from NHEJ to HR. BRCA1 and POH1 act as distinct but interfacing barriers to ubiquitin chain removal.","method":"siRNA co-depletion experiments, immunofluorescence for DSB factors (53BP1, RAP80, RPA, ubiquitin FK2), cell cycle-staged analysis","journal":"Nucleic Acids Research","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple co-depletion epistasis experiments, specific molecular readouts, pathway position defined","pmids":["24013561"],"is_preprint":false},{"year":2014,"finding":"Crystal structures of the Rpn11-Rpn8 heterodimer at 2.0 Å resolution reveal that Rpn11 lacks a conserved surface for ubiquitin Ile44-patch binding, does not contact the proximal side of the scissile isopeptide bond, and exhibits no ubiquitin linkage specificity. Two distinct interfaces mediate the Rpn11-Rpn8 interaction. Mutational studies confirm these structural features explain how Rpn11 functions as a promiscuous, cotranslocational deubiquitinase.","method":"X-ray crystallography (Zn2+-free and Zn2+-bound structures), site-directed mutagenesis, biochemical DUB assays","journal":"Nature Structural & Molecular Biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure + mutagenesis + functional validation, replicated by independent structure paper (PMID:24516147)","pmids":["24463465"],"is_preprint":false},{"year":2014,"finding":"Crystal structures of the Rpn8-Rpn11 MPN-domain heterodimer (obtained via nanobody-assisted crystallization) reveal that full Rpn11 activation requires incorporation into the 26S proteasome and is dependent on ATP hydrolysis. Premature activation is prevented by: (1) low intrinsic ubiquitin affinity, (2) an insertion segment acting as a physical barrier across the substrate access channel, and (3) a conformationally unstable catalytic loop. Docking into proteasome EM density shows Rpn11 contacts ATPase subunits that stabilize the active conformation.","method":"X-ray crystallography (three structures), cryo-EM density docking, nanobody-assisted crystallization, DUB activity assay with model substrate","journal":"PNAS","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure with functional validation, complementary to PMID:24463465","pmids":["24516147"],"is_preprint":false},{"year":2014,"finding":"Rpn11 and Ubp6 process K11- and K63-linked ubiquitin chains with comparable efficiencies (increasing with chain length), whereas proteasomal processing of K48-linked chains is inversely correlated with chain length. Incorporation into proteasomes enhances Rpn11 enzymatic efficiency by roughly 2 orders of magnitude, partly by relieving autoinhibition by its C-terminus. Rpn11 shows a random cleavage mode on K48 chains (broad/endo), while Ubp6 shows endo-chain preference on K48.","method":"In vitro DUB assays with defined ubiquitin conjugates (homogeneous K11, K48, K63 chains of varying lengths, heterogeneous chains), fluorescently labeled Ub chains, purified proteasomes vs isolated DUBs","journal":"Journal of Biological Chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — rigorous in vitro biochemistry with panel of defined substrates, single lab but multiple orthogonal assays","pmids":["25389291"],"is_preprint":false},{"year":2015,"finding":"POH1/PSMD14 deubiquitylates and stabilizes E2F1. POH1 physically binds E2F1 and removes ubiquitin from it, preventing its proteasomal degradation. Conditional knockout of Poh1 in primary mouse liver cells reduces E2F1 protein levels. Stabilized E2F1 upregulates Survivin and FOXM1, promoting liver cancer cell tumor growth.","method":"Co-immunoprecipitation, in vivo ubiquitination assays, conditional Poh1 knockout mouse model, Western blot, xenograft mouse model","journal":"Nature Communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, in vivo ubiquitination assay, conditional KO mouse, replicated in multiple cancer-type papers","pmids":["26510456"],"is_preprint":false},{"year":2017,"finding":"Rpn11 DUB activity is coupled to substrate translocation by the AAA+ ATPase motor via a conformational switch of the Insert-1 (Ins-1) loop. Ubiquitin binding induces Ins-1 transition from an inactive closed state to an active β-hairpin; this switch is rate-limiting for deubiquitination and is strongly accelerated by mechanical substrate translocation. Deubiquitination by Rpn11 and ubiquitin unfolding by the ATPases are in direct competition, requiring rapid Rpn11 activation to prevent ubiquitin co-degradation.","method":"X-ray crystallography (ubiquitin-bound Rpn11 structure), mutagenesis, in vitro single-molecule and ensemble DUB assays, biochemical translocation-deubiquitination coupling assays","journal":"Molecular Cell","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure of ubiquitin-bound state, mutagenesis, multiple biochemical assays establishing mechanochemical coupling","pmids":["28844860"],"is_preprint":false},{"year":2017,"finding":"Quinoline-8-thiol derivative capzimin is a selective inhibitor of proteasomal Rpn11/PSMD14 that inhibits the JAMM metalloprotease by chelating its active-site zinc. Capzimin (>5-fold selectivity for Rpn11 over related JAMM proteases) stabilizes polyubiquitinated proteasome substrates, induces unfolded protein response, and blocks cancer cell proliferation including bortezomib-resistant cells.","method":"Biochemical DUB activity assays, proteomic analysis of stabilized substrates, cell viability assays, selectivity profiling against related JAMM proteases and metalloenzymes","journal":"Nature Chemical Biology","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — biochemical characterization, selectivity profiling, cellular proteomics, multiple cell line validation","pmids":["28244987"],"is_preprint":false},{"year":2017,"finding":"Thiolutin, a disulfide-containing antibiotic, is a zinc chelator that inhibits the JAMM metalloprotease Rpn11/PSMD14 of the 19S proteasome in its reduced form. It also inhibits related JAMM metalloproteases Csn5, AMSH, and BRCC36.","method":"Biochemical DUB inhibition assays, zinc chelation characterization, zinc-binding experiments with Rpn11 and related JAMM enzymes","journal":"Nature Chemical Biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro biochemical inhibition assays across multiple JAMM family members, zinc chelation mechanism established","pmids":["28459440"],"is_preprint":false},{"year":2018,"finding":"PSMD14 deubiquitylates SNAIL (EMT transcription factor), preventing its ubiquitin-mediated proteasomal degradation, thereby stabilizing SNAIL protein. Mass spectrometry identified PSMD14 as a SNAIL-interacting DUB. PSMD14 knockdown blocks SNAIL-induced EMT, suppresses tumor cell migration and invasion in vitro, and inhibits metastasis in vivo.","method":"Mass spectrometry interactome screen, co-immunoprecipitation, in vivo ubiquitination assay, siRNA knockdown with migration/invasion assays, xenograft metastasis model","journal":"Cancer Letters","confidence":"High","confidence_rationale":"Tier 2 / Moderate — MS-identified interaction, reciprocal Co-IP, in vivo ubiquitination, functional in vivo validation","pmids":["29331416"],"is_preprint":false},{"year":2018,"finding":"Epidithiodiketopiperazines (ETPs) inhibit proteasomal degradation by targeting Rpn11/POH1/PSMD14, the essential proteasomal DUB. ETPs also inhibit related JAMM proteases Csn5 and AMSH. An improved ETP (SOP11) stabilizes a subset of polyubiquitinated proteasome substrates, induces the unfolded protein response, and causes cell death.","method":"In vitro reconstituted proteasome-mediated protein degradation assay, biochemical Rpn11 inhibition assays, cellular proteasome substrate stabilization, UPR reporter assays","journal":"Cell Chemical Biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstituted degradation assay plus cellular validation, single lab","pmids":["30146242"],"is_preprint":false},{"year":2018,"finding":"POH1/PSMD14 deubiquitinates pro-IL-1β by removing K63-linked polyubiquitin chains, decreasing its susceptibility to cleavage and mature IL-1β production. POH1 physically interacts with pro-IL-1β. Myeloid cell-specific POH1 deletion aggravates LPS-induced systemic inflammation and alum-induced peritonitis in vivo.","method":"Co-immunoprecipitation, K63-specific deubiquitination assay, myeloid-specific conditional knockout mice, in vivo inflammation models (LPS, alum peritonitis), IL-1β production measurement","journal":"Nature Communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — Co-IP, linkage-specific ubiquitination assay, conditional KO mouse with in vivo phenotype","pmids":["30315153"],"is_preprint":false},{"year":2019,"finding":"PSMD14 deubiquitinates and stabilizes GRB2 via inhibiting its ubiquitin-mediated proteasomal degradation, promoting HCC proliferation, migration, and invasion.","method":"Co-immunoprecipitation, in vivo ubiquitination assay, siRNA knockdown, overexpression experiments, xenograft and metastasis mouse models","journal":"Cancer Letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and ubiquitination assay, in vivo validation, single lab","pmids":["31634528"],"is_preprint":false},{"year":2019,"finding":"PSMD14 deubiquitinates the ALK2 (BMP type I receptor) by removing K48-linked ubiquitin chains added by Smurf1 E3 ligase, thereby stabilizing ALK2 and promoting BMP6 signaling pathway activation. This function is stated to be independent of its intrinsic role in the 26S proteasome. PSMD14 was identified via siRNA DUB library screen.","method":"siRNA DUB library screen, immunoblot, co-immunoprecipitation, K48-specific ubiquitination assays, xenograft colorectal cancer model","journal":"EBioMedicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — DUB screen, Co-IP, linkage-specific ubiquitination assay, in vivo validation, single lab","pmids":["31685442"],"is_preprint":false},{"year":2019,"finding":"POH1 deubiquitinates TGF-β receptors (TGFBR1 and TGFBR2) and caveolin-1 (CAV1), preventing lysosome pathway-mediated turnover of TGF-β receptors and thereby hyperactivating TGF-β signaling. POH1-deficient mouse hepatocytes show severely downregulated TGF-β receptor levels. This promotes HCC metastatic properties in vitro and in vivo.","method":"Western blotting, co-immunoprecipitation, ubiquitination assays, conditional Poh1 knockout mouse (Mx-Cre+, poh1f/f), xenograft and metastasis models","journal":"EBioMedicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — Co-IP, ubiquitination assay, conditional KO mouse with in vivo phenotype, multiple substrates investigated","pmids":["30745168"],"is_preprint":false},{"year":2020,"finding":"PSMD14 depletion or pharmacological inhibition (capzimin) causes retention of Atg9A and Rab1A at the Golgi apparatus, blocking Golgi-to-ER retrograde transport and consequently inhibiting macroautophagy. PSMD14 acts specifically on K63-linked ubiquitin chains in cells. Inhibition of the 20S proteasome did not recapitulate these trafficking effects, indicating a specific role for PSMD14/K63-Ub in Golgi-to-ER retrograde transport.","method":"High-content siRNA screening (1187 ubiquitinome genes), APP trafficking reporter, siRNA knockdown, capzimin treatment, fluorescence microscopy for Golgi/ER trafficking markers, autophagy flux assays","journal":"Cells","confidence":"High","confidence_rationale":"Tier 2 / Strong — genome-scale siRNA screen, pharmacological validation, specific trafficking assays, 20S vs 19S comparison as control","pmids":["32210007"],"is_preprint":false},{"year":2021,"finding":"Arsenite inhibits PSMD14/Rpn11 metalloprotease DUB activity by substituting zinc in the MPN/JAMM domain. The proteasomal adaptor AIRAP can directly relieve this PSMD14/Rpn11 inhibition, suggesting a metal relay mechanism between arsenylated PSMD14 and AIRAP to restore proteasomal DUB function during arsenite stress.","method":"Direct arsenite binding assay, in vitro Rpn11 DUB inhibition assay, AIRAP interaction and relief-of-inhibition experiments","journal":"Biomolecules","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — biochemical inhibition and relief assays, single lab, mechanistic model proposed but limited orthogonal validation","pmids":["34572530"],"is_preprint":false},{"year":2021,"finding":"PSMD14 decreases K63-linked ubiquitination on PKM2, shifts the PKM2 oligomeric equilibrium from tetramers toward dimers/monomers, diminishes pyruvate kinase enzymatic activity, and induces nuclear translocation of PKM2, thereby promoting aerobic glycolysis in ovarian cancer cells.","method":"Co-immunoprecipitation, K63-specific ubiquitination assays, PKM2 oligomeric state analysis (native PAGE), pyruvate kinase activity assay, nuclear fractionation, siRNA knockdown/overexpression","journal":"Molecular Oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — linkage-specific ubiquitination, enzymatic activity assay, fractionation, single lab","pmids":["34382324"],"is_preprint":false},{"year":2022,"finding":"PSMD14 deubiquitinates and stabilizes LRPPRC by inhibiting its ubiquitination, thereby suppressing autophagy through the LRPPRC/Beclin1-Bcl-2/SQSTM1 signaling pathway in ovarian cancer cells.","method":"Co-immunoprecipitation, ubiquitination assays, autophagy flux assays, siRNA knockdown, xenograft and metastasis mouse models","journal":"BBA Molecular Basis of Disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assay, pathway analysis, in vivo validation, single lab","pmids":["36328147"],"is_preprint":false},{"year":2023,"finding":"PSMD14 acts as a histone H2AK119 deubiquitinase independently of the 19S regulatory particle, functioning on chromatin in complex with NSD2. This non-proteasomal PSMD14 activity facilitates NSD2-directed H3K36 dimethylation and transcriptional activation of target genes including RELA, driving myelomagenesis. RELA in turn transactivates PSMD14, forming a positive feedback loop.","method":"ChIP-seq, co-immunoprecipitation (chromatin-bound PSMD14-NSD2 complex), histone H2AK119 deubiquitination assays, integrative genomic/epigenomic analyses, PSMD14 inhibitor studies","journal":"Molecular Cell","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — chromatin biochemistry, histone DUB assay, integrative epigenomics, functional feedback loop demonstrated","pmids":["37935198"],"is_preprint":false},{"year":2023,"finding":"POH1 deubiquitinates and stabilizes the MYC protein, which potentiates acinar-to-ductal metaplasia (ADM) and pancreatic ductal adenocarcinoma (PDAC). Pancreatic-specific deletion of Poh1 attenuates ADM and impairs pancreatic carcinogenesis in murine models.","method":"Pancreatic-specific conditional Poh1 knockout mouse, co-immunoprecipitation, in vivo ubiquitination assay, ADM murine models","journal":"Cancer Letters","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional KO mouse with in vivo carcinogenesis phenotype, Co-IP, ubiquitination assay","pmids":["37844756"],"is_preprint":false},{"year":2023,"finding":"POH1 directly interacts with Smad3, removes poly-ubiquitin modifications from Smad3, stabilizes it, and thereby facilitates TGF-β1-mediated lung cancer cell invasion and metastasis.","method":"Co-immunoprecipitation, colocalization analysis, in vitro deubiquitination, half-life assay, functional migration/invasion assays, xenograft liver metastasis model","journal":"Cancer Letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, in vitro deubiquitination, functional rescue, single lab","pmids":["38061486"],"is_preprint":false},{"year":2023,"finding":"PSMD14 stabilizes ERα by removing K48-linked polyubiquitin chains, thereby maintaining ERα protein levels and ERα transcriptome in breast cancer. ERα reciprocally binds the PSMD14 promoter to promote its transcription, forming a positive feedback loop. In endocrine-resistant models, PSMD14 inhibition destabilizes the resistant ERα Y537S mutant and restores tamoxifen sensitivity.","method":"siRNA DUB library screen, co-immunoprecipitation, K48-specific ubiquitination assay, ChIP assay, endocrine-resistance model, in vivo xenograft","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Strong — DUB screen, linkage-specific ubiquitination, ChIP, resistant mutant model, in vivo validation","pmids":["38017133"],"is_preprint":false},{"year":2024,"finding":"PSMD14 deubiquitinates β-catenin by decreasing its K48-linked ubiquitination, preventing β-catenin proteasomal degradation and stabilizing it, thereby promoting GBM cell proliferation and invasion. PSMD14 directly interacts with β-catenin.","method":"Co-immunoprecipitation, K48-specific ubiquitination assay, siRNA knockdown, rescue with β-catenin overexpression, xenograft mouse model","journal":"BioFactors","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, linkage-specific ubiquitination, functional rescue, single lab","pmids":["38696072"],"is_preprint":false},{"year":2024,"finding":"RPN11/PSMD14 deubiquitinates and stabilizes METTL3, an m6A RNA methyltransferase. Stabilized METTL3 enhances m6A modification and expression of ACSS3, which generates propionyl-CoA to upregulate lipid metabolism genes via histone propionylation. Hepatocyte-specific RPN11 knockout mice are protected from diet-induced liver steatosis, insulin resistance, and steatohepatitis.","method":"Hepatocyte-specific conditional RPN11 knockout mice, co-immunoprecipitation, ubiquitination assays, m6A sequencing, metabolomics, in vivo diet-induced NAFLD models, pharmacological inhibition (capzimin)","journal":"Cell Metabolism","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional KO mouse, Co-IP, ubiquitination assay, metabolomics, m6A sequencing, pharmacological validation","pmids":["39146936"],"is_preprint":false},{"year":2025,"finding":"Rpn11 functions as an allosteric ubiquitin sensor at the 26S proteasome. After substrate recruitment, ubiquitin binding to Rpn11 interferes with conformation-specific interactions of the ubiquitin receptor Rpn10, thereby stabilizing the engagement-competent proteasome state and expediting substrate insertion into the ATPase motor. This allosteric mechanism allows poly-ubiquitin chains or multiple mono-ubiquitins to promote up to 4-fold faster proteasomal turnover.","method":"Biochemical assays, active-site and interface mutagenesis, single-molecule FRET (smFRET) measurements of proteasome conformational states, reconstituted degradation assays","journal":"Cell Reports","confidence":"High","confidence_rationale":"Tier 1 / Strong — smFRET, mutagenesis, reconstituted biochemistry, multiple orthogonal methods establishing allosteric mechanism","pmids":["40411784"],"is_preprint":false},{"year":2025,"finding":"PSMD14 prevents SLC7A11 ubiquitination and proteasomal degradation by binding to it; glucocorticoids impede this PSMD14-SLC7A11 interaction, causing SLC7A11 degradation, cystine insufficiency, and osteocyte ferroptosis in GIOP. Bone-targeting AAV-mediated PSMD14 overexpression stabilizes SLC7A11 and attenuates both osteocyte ferroptosis and bone loss.","method":"Co-immunoprecipitation, ubiquitination assays, AAV-mediated overexpression in mouse model, GPX4 conditional KO, pharmacological ferroptosis inhibitors, high-throughput virtual screening","journal":"Advanced Science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assay, in vivo mouse model, single lab","pmids":["40444470"],"is_preprint":false},{"year":2025,"finding":"PSMD14 directly binds and deubiquitinates LDHA to stabilize it, increasing intracellular lactate, which elevates histone lactylation (H3K18la) to transcriptionally activate ACLY and promote lipid biosynthesis and pancreatic cancer progression.","method":"Co-immunoprecipitation, ubiquitination assays, histone lactylation measurements, ACLY expression analysis, patient-derived xenograft (PDX) models","journal":"Advanced Science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assay, epigenetic readout, PDX validation, single lab","pmids":["41051446"],"is_preprint":false},{"year":2025,"finding":"PSMD14 directly deubiquitinates BCKDK, antagonizing TRIM21-mediated proteasomal degradation, thereby stabilizing BCKDK and promoting SLC7A5/SLC7A8-mediated branched-chain amino acid (BCAA) uptake by GBM cells, leading to NK cell BCAA depletion and immune suppression.","method":"Co-immunoprecipitation, ubiquitination assays, PSMD14 knockdown/pharmacological inhibition (OPA), NK cytotoxicity assays, preclinical GBM models","journal":"Cell Death & Differentiation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assay, functional immune cell assays, in vivo validation, single lab","pmids":["41876842"],"is_preprint":false},{"year":2025,"finding":"PSMD14 stabilizes IMPDH2, the rate-limiting enzyme of purine nucleotide biosynthesis, by selectively removing K48-linked polyubiquitin chains. PSMD14 inhibition decreases IMPDH2 stability, impairs nucleotide metabolism, causes mitochondrial dysfunction, and increases DNA damage signaling in GBM. Exogenous guanosine reverses these effects.","method":"Immunoprecipitation-coupled mass spectrometry (IP-MS), K48-specific ubiquitination assays, metabolic assays, mitochondrial function assessments, guanosine rescue experiments, orthotopic GBM mouse models","journal":"Theranostics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — IP-MS, linkage-specific ubiquitination, metabolic rescue experiment, in vivo validation, single lab","pmids":["41608571"],"is_preprint":false},{"year":2025,"finding":"PSMD14 stabilizes PD-L1 by interacting with its intracellular domain and removing K48-linked polyubiquitin chains, thereby inhibiting proteasomal degradation of PD-L1. PSMD14 inhibition promotes PD-L1 degradation, enhances CD8+ T cell activation, reduces Tregs and MDSCs, and improves immunotherapy response in breast cancer syngeneic models.","method":"Co-immunoprecipitation, K48-specific ubiquitination assay, cycloheximide chase assay, flow cytometry, T cell cytotoxicity assay, syngeneic mouse models","journal":"Journal of Experimental & Clinical Cancer Research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, linkage-specific ubiquitination, CHX chase, in vivo syngeneic model, single lab","pmids":["41981629"],"is_preprint":false},{"year":2025,"finding":"PSMD14 deubiquitinates SF3B4 (K63-linked ubiquitin removal), stabilizing SF3B4, which then forms a complex with HNRNPC to promote exon inclusion in FADS1 mRNA via m6A-HNRNPC recognition, upregulating FADS1 and activating Akt/mTOR signaling in TNBC.","method":"Co-IP, K63-specific deubiquitination assays, RNA splicing analysis, m6A-RIP, TNBC organoid (PDO) and PDX models","journal":"Science Advances","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, linkage-specific ubiquitination, RNA mechanistic assays, PDO/PDX validation, single lab","pmids":["40344056"],"is_preprint":false},{"year":2025,"finding":"TXNL1's C-terminal tail covers the catalytic groove of the Rpn11 deubiquitinase and coordinates the active-site Zn2+ specifically in substrate-degrading proteasome conformational states (but not in resting state), as revealed by time-resolved cryo-EM. This is consistent with TXNL1 binding to the actively processing proteasome and potentially modulating Rpn11 activity.","method":"Time-resolved cryo-EM at saturating and sub-stoichiometric TXNL1 concentrations, biophysical binding experiments","journal":"bioRxiv (preprint)","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — high-resolution cryo-EM structure, preprint not yet peer-reviewed, single lab","pmids":["bio_10.1101_2024.11.08.622731"],"is_preprint":true},{"year":2025,"finding":"PSMD14/Rpn11 functions non-enzymatically as a receptor for the midnolin ubiquitin-like (Ubl) domain in the midnolin-proteasome pathway (ubiquitin-independent degradation). Cryo-EM structures of the MIDN-bound proteasome show the midnolin Ubl domain binding to Rpn11 (not cleaved by it), positioning the substrate-binding Catch domain above the proteasomal channel. This is distinct from Rpn11's canonical deubiquitinase function.","method":"Cryo-EM structures of MIDN-bound human proteasome in two conformational states","journal":"bioRxiv (preprint)","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — cryo-EM structure, preprint, corroborated by independent parallel preprint (bio_10.1101_2025.03.04.641557)","pmids":["bio_10.1101_2025.02.22.639686"],"is_preprint":true},{"year":2025,"finding":"In the ubiquitin-independent ODC degradation pathway, Rpn11's JAMM motif guides the ODC C-tail into the AAA+ ATPase ring, acting as a translocation gateway and repurposing Rpn11 for a novel ubiquitin-independent function distinct from its canonical deubiquitinase role.","method":"Cryo-EM (eleven structures of human 26S proteasome-ODC complexes capturing full degradation process), mutagenesis","journal":"bioRxiv (preprint)","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — multi-state cryo-EM capturing complete substrate processing, structural basis for non-canonical function, preprint","pmids":["bio_10.1101_2025.11.15.688597"],"is_preprint":true},{"year":2025,"finding":"PSMD14 deubiquitinates CARM1 (coactivator-associated arginine methyltransferase 1), stabilizing it. Stabilized CARM1 activates transcription of FERMT1 through H3R17 dimethylation, promoting HCC proliferation and metastasis.","method":"Co-immunoprecipitation, ubiquitination assays, ChIP assay, in vitro and in vivo cell proliferation/metastasis models","journal":"Cell Death & Disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assay, ChIP, in vivo validation, single lab","pmids":["40016178"],"is_preprint":false},{"year":2025,"finding":"PSMD14 deubiquitinates RBM15B, preventing its ubiquitin-mediated degradation. Stabilized RBM15B promotes m6A modification of SPON2 mRNA and its stability, facilitating pancreatic cancer progression. PSMD14 itself is transcriptionally activated by MEF2A.","method":"ChIP assay (MEF2A-PSMD14 promoter), dual luciferase, co-immunoprecipitation, ubiquitination assay, RIP assay (RBM15B-SPON2 mRNA), MeRIP (m6A on SPON2), subcutaneous and lung metastasis mouse models","journal":"Kaohsiung Journal of Medical Sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple molecular assays, in vivo validation, single lab","pmids":["40066751"],"is_preprint":false},{"year":2025,"finding":"PSMD14 deubiquitinates PFKFB2 at K355, facilitating SCYL2-mediated phosphorylation of PFKFB2, which increases fructose-2,6-bisphosphate generation, activates PFK1, and promotes glycolysis and H3K27 lactylation in gastric adenocarcinoma. H3K27 lactylation in turn activates PSMD14 and SOX9 expression, forming a positive feedback loop.","method":"Co-immunoprecipitation, site-specific ubiquitination assay (K355), phosphorylation assays, glycolysis metabolic assays, H3K27 lactylation ChIP, FDA drug screening for PSMD14 inhibitors","journal":"Cell Death & Differentiation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — site-specific ubiquitination, metabolic assays, epigenetic readout, single lab","pmids":["41193870"],"is_preprint":false},{"year":2025,"finding":"PSMD14 stabilizes FOXM1 by reducing K63-linked ubiquitination on FOXM1, promoting breast cancer progression. PSMD14 also activates the PI3K/AKT/mTOR pathway.","method":"Co-immunoprecipitation, immunofluorescence, K63-specific in vitro and in vivo deubiquitination assays, xenograft mouse model","journal":"International Journal of Biological Macromolecules","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, linkage-specific ubiquitination, in vivo validation, single lab","pmids":["40902741"],"is_preprint":false},{"year":2025,"finding":"PSMD14 deubiquitinates and stabilizes CDC42 in endothelial cells, promoting filopodia formation and cell migration. Silencing PSMD14 impairs filopodia formation, migration, and CDC42 stability.","method":"Co-immunoprecipitation (PSMD14-CDC42 interaction), CDC42 stability/ubiquitination assays, siRNA knockdown, filopodia imaging, migration assays, mouse hindlimb ischemia model","journal":"Angiogenesis","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, functional loss-of-function, in vivo model, single lab","pmids":["40833626"],"is_preprint":false},{"year":2025,"finding":"Histone lactylation (H3K18la) upregulates PSMD14 transcription after traumatic brain injury. PSMD14 then deubiquitinates PKM2, maintaining PKM protein stability and enabling PSMD14-mediated mitophagy via PINK1 phosphorylation at Thr257, thereby suppressing neuron PANoptosis.","method":"LC-MS proteomic analysis, co-immunoprecipitation, ubiquitination assays, PINK1 phosphorylation assays, mitophagy flux analysis, controlled cortical impact (CCI) mouse model","journal":"Autophagy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assay, in vivo CCI model, mechanistic pathway validated, single lab","pmids":["40000916"],"is_preprint":false},{"year":2026,"finding":"DUSP4-mediated dephosphorylation of PSMD14 enhances PSMD14's interaction with MICALL2 and its deubiquitination activity toward MICALL2, stabilizing MICALL2 and promoting ccRCC malignant progression.","method":"Co-immunoprecipitation, MICALL2 ubiquitination assay, PSMD14 phosphorylation analysis, siRNA knockdown, in vivo ccRCC mouse model","journal":"Journal of Translational Medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination and phosphorylation assays, in vivo model, single lab","pmids":["42174657"],"is_preprint":false}],"current_model":"PSMD14/POH1/Rpn11 is an essential zinc-dependent JAMM-family metalloprotease deubiquitinase that resides in the lid subcomplex of the 19S proteasome regulatory particle, where it couples substrate deubiquitination to ATP-dependent degradation by cleaving poly-ubiquitin chains en bloc at the base upon mechanical substrate translocation; its catalytic activity requires an intact JAMM motif and is allosterically regulated by ubiquitin binding (promoting proteasome engagement) and by ATPase-driven conformational switching; beyond the proteasome, PSMD14 deubiquitinates a wide array of non-histone substrates (e.g., E2F1, SNAIL, GRB2, TGF-β receptors, pro-IL-1β, ErbB2, ERα, β-catenin, METTL3, PKM2, ALK2, MYC, LDHA, Smad3) to regulate their stability independently of degradation, controls K63-linked ubiquitin chains at DNA double-strand break sites to regulate DNA repair pathway choice, acts as an epigenetic H2AK119 deubiquitinase on chromatin with NSD2, and regulates Golgi-to-ER retrograde transport and macroautophagy."},"narrative":{"mechanistic_narrative":"PSMD14 (POH1/Rpn11) is the essential zinc-dependent JAMM-family metalloprotease deubiquitinase of the 19S proteasome lid that couples substrate deubiquitination to ATP-dependent proteolysis, removing poly-ubiquitin chains en bloc as substrates are translocated into the degradation machinery [PMID:12183636, PMID:14581483]. Its catalytic activity resides in the MPN+/JAMM motif, whose conserved active-site residues are required for viability in yeast and human cells and for proteasomal degradation; active-site mutation abolishes deubiquitination and is lethal [PMID:12183636, PMID:12370088, PMID:17237285]. Structural and biochemical work established that Rpn11 is a promiscuous, low-affinity deubiquitinase lacking ubiquitin-linkage specificity at the level of the isolated MPN domain, and that full activation requires incorporation into the 26S proteasome and ATP hydrolysis—premature activity being prevented by an Insert-1 barrier loop and an unstable catalytic loop [PMID:24463465, PMID:24516147]. Substrate-driven ubiquitin binding switches the Ins-1 loop to an active conformation in a step that is rate-limiting and accelerated by mechanical translocation, placing Rpn11 in direct kinetic competition with ATPase-driven ubiquitin unfolding [PMID:28844860]; ubiquitin binding also acts allosterically through Rpn10 to stabilize the engagement-competent proteasome state and speed turnover [PMID:40411784]. Within and beyond the proteasome, PSMD14 preferentially processes K63-linked chains [PMID:19214193, PMID:25389291], a specificity it exploits at sites of DNA double-strand breaks, where it removes RNF8/RNF168-generated K63 chains to limit 53BP1 accumulation, counter the RAP80 barrier to end resection, and promote homologous recombination [PMID:22909820, PMID:24013561]. Loss of PSMD14 causes polyubiquitin accumulation, G0/G1 arrest and senescence, distinguishing 19S from 20S function [PMID:17237285, PMID:19732767]. Independently of bulk degradation, PSMD14 deubiquitinates and stabilizes numerous substrates—E2F1, SNAIL, TGF-β receptors and Smad3, ALK2, ERα, β-catenin, MYC, GRB2 and others—removing K48- or K63-linked chains to control their stability and downstream signaling, frequently in cancer contexts and often within transcriptional positive-feedback loops [PMID:26510456, PMID:29331416, PMID:30745168, PMID:31685442, PMID:38017133, PMID:37844756, PMID:38061486]. PSMD14 further acts on chromatin as a non-proteasomal H2AK119 deubiquitinase in complex with NSD2 to license H3K36 methylation and transcription [PMID:37935198], regulates K63-dependent Golgi-to-ER retrograde transport and macroautophagy [PMID:32210007], and is targeted by zinc-chelating inhibitors including capzimin and thiolutin that stabilize proteasome substrates and induce the unfolded protein response [PMID:28244987, PMID:28459440].","teleology":[{"year":2002,"claim":"Establishing whether the proteasome lid contains an intrinsic deubiquitinase resolved how ubiquitin is removed during degradation: Rpn11 was identified as a JAMM metalloprotease whose activity is mechanistically coupled to substrate proteolysis.","evidence":"Active-site mutagenesis (rpn11AXA) with yeast lethality and in vitro reconstituted degradation of ubiquitinated Sic1; MPN+ motif mutagenesis with proteolysis defects","pmids":["12183636","12370088"],"confidence":"High","gaps":["Did not resolve the structural basis of substrate engagement","Linkage specificity not yet defined"]},{"year":2003,"claim":"Defining how proteasomal deubiquitination is partitioned showed Rpn11 and Ubp6 play complementary, non-redundant roles and confirmed Rpn11 as a metal-dependent enzyme requiring an intact lid for coupling to degradation.","evidence":"Proteasome purification from yeast mutants, in vitro DUB assays, metal chelator sensitivity, synthetic lethality genetics","pmids":["14581483"],"confidence":"High","gaps":["Mechanism coupling deubiquitination to translocation not defined","Metal identity inferred, not structurally resolved"]},{"year":2004,"claim":"Dissecting Rpn11 into separable domains revealed a catalytic-independent C-terminal function in mitochondrial morphology, indicating roles beyond proteasomal DUB activity.","evidence":"Yeast RPN8-RPN11 chimera analysis, intragenic complementation, mitochondrial microscopy, cell cycle analysis","pmids":["15018611"],"confidence":"High","gaps":["Molecular mechanism of the mitochondrial function unresolved","Not extended to mammalian cells in this work"]},{"year":2007,"claim":"Testing requirement in human cells confirmed the JAMM motif is essential for viability and 26S function, validating PSMD14 catalysis as the operative human activity.","evidence":"siRNA knockdown with wild-type vs JAMM-mutant rescue, viability and polyubiquitin accumulation assays in HeLa cells","pmids":["17237285"],"confidence":"High","gaps":["Did not address non-proteasomal functions","Substrate scope not examined"]},{"year":2009,"claim":"Defining linkage specificity and downstream phenotype showed proteasome-associated PSMD14 is a K63-specific metalloDUB and that 19S loss produces cell cycle arrest distinct from 20S inhibition.","evidence":"Seven-step chromatographic co-fractionation with linkage-specific assays and inhibitor profiling; siRNA with flow cytometry and comparison to PSMB5","pmids":["19214193","19732767"],"confidence":"High","gaps":["K63 specificity at the isolated enzyme level not yet reconciled with later structural promiscuity","Direct substrates driving arrest not identified"]},{"year":2009,"claim":"Early substrate studies hinted that PSMD14 deubiquitinates specific proteins (c-Jun, ErbB2) in a manner not strictly coupled to degradation, foreshadowing degradation-independent stabilization roles.","evidence":"Ectopic overexpression and active-site mutagenesis for c-Jun; siRNA DUB screen and Western/flow cytometry for ErbB2","pmids":["16569633","19436748"],"confidence":"Medium","gaps":["Single-lab Co-IP/ubiquitination without reciprocal validation","Selectivity claims rest on limited substrate panels"]},{"year":2013,"claim":"Positioning PSMD14 in DNA repair established that it processes K63 chains at break sites to limit 53BP1, counter the RAP80 resection barrier, and bias repair toward homologous recombination.","evidence":"siRNA knockdown and co-depletion, immunofluorescence of DSB foci, K63-Ub chromatin analysis, survival assays","pmids":["22909820","24013561"],"confidence":"High","gaps":["Whether this is proteasome-dependent or free-DUB activity not fully resolved","Direct chromatin substrates not enumerated"]},{"year":2014,"claim":"Crystallographic and biochemical analyses resolved the structural logic of Rpn11: it is an intrinsically promiscuous, low-affinity DUB whose activation is gated by proteasome incorporation and ATP hydrolysis.","evidence":"Rpn11-Rpn8 heterodimer crystal structures, nanobody-assisted crystallization, cryo-EM docking, mutagenesis, defined-chain in vitro DUB kinetics","pmids":["24463465","24516147","25389291"],"confidence":"High","gaps":["The ubiquitin-bound active conformation not yet captured at this stage","Reconciliation with cellular K63 preference incomplete"]},{"year":2015,"claim":"An in vivo substrate paradigm showed PSMD14 stabilizes E2F1 to drive oncogenic transcription, exemplifying degradation-independent substrate control in mammals.","evidence":"Reciprocal Co-IP, in vivo ubiquitination, conditional Poh1 knockout mouse liver, xenografts","pmids":["26510456"],"confidence":"High","gaps":["Whether stabilization is proteasome-associated or free-enzyme not dissected","Linkage type on E2F1 not specified"]},{"year":2017,"claim":"Capturing the ubiquitin-bound state and developing selective inhibitors defined the mechanochemical activation switch and validated PSMD14 as a druggable target competing with ATPase-driven ubiquitin unfolding.","evidence":"Ubiquitin-bound Rpn11 crystal structure with single-molecule and ensemble DUB assays; capzimin and thiolutin zinc-chelation inhibition with proteomics and cell viability","pmids":["28844860","28244987","28459440"],"confidence":"High","gaps":["Inhibitor selectivity over related JAMM proteases is limited","In vivo therapeutic window not established in these studies"]},{"year":2019,"claim":"Expanding the substrate repertoire established broad degradation-independent stabilization activity across SNAIL, TGF-β receptors, ALK2 and GRB2, frequently via removal of specific K48 or K63 chains in cancer.","evidence":"Mass spectrometry interactomics, Co-IP, linkage-specific ubiquitination assays, conditional knockout mice, xenograft and metastasis models","pmids":["29331416","30315153","30745168","31634528","31685442"],"confidence":"High","gaps":["Most substrate studies are single-lab without reciprocal cross-validation","Whether substrate DUB activity requires proteasome incorporation often untested"]},{"year":2020,"claim":"A trafficking and autophagy role showed PSMD14 acts on cellular K63 chains to enable Golgi-to-ER retrograde transport, distinct from 20S proteasome function.","evidence":"High-content siRNA ubiquitinome screen, capzimin treatment, trafficking-marker microscopy, autophagy flux, 19S vs 20S comparison","pmids":["32210007"],"confidence":"High","gaps":["Direct K63-ubiquitinated trafficking substrates not identified","Mechanism of recruitment to Golgi unresolved"]},{"year":2023,"claim":"Demonstrating a chromatin-resident, proteasome-independent activity revealed PSMD14 as an H2AK119 deubiquitinase in an NSD2 complex that licenses transcription, establishing a non-proteasomal epigenetic role.","evidence":"ChIP-seq, chromatin Co-IP, histone H2AK119 deubiquitination assays, integrative epigenomics, inhibitor studies","pmids":["37935198"],"confidence":"High","gaps":["How PSMD14 is partitioned between proteasome and chromatin pools unclear","Generality across cell types beyond myeloma not established"]},{"year":2025,"claim":"Mechanistic refinement defined PSMD14 as an allosteric ubiquitin sensor that accelerates proteasomal turnover via Rpn10, while a wave of substrate studies (MYC, ERα, METTL3, LDHA, β-catenin, PD-L1, BCKDK, IMPDH2, metabolic and immune regulators) extended its degradation-independent stabilization activity into metabolism, immunity, and feedback transcriptional loops.","evidence":"smFRET, interface mutagenesis and reconstituted degradation for the allosteric mechanism; Co-IP, linkage-specific ubiquitination, conditional knockouts and PDX/syngeneic models for substrates","pmids":["40411784","37844756","38017133","39146936","41051446","38696072","41981629","41876842","41608571"],"confidence":"High","gaps":["Substrate studies are predominantly single-lab and context-specific","Whether individual substrate DUB events occur at the proteasome or by a free enzyme pool largely untested"]},{"year":2025,"claim":"Structural studies began to reveal non-canonical, ubiquitin-independent uses of Rpn11—as a midnolin Ubl receptor and an ODC translocation gateway—and a TXNL1-mediated regulatory contact in processing-state proteasomes.","evidence":"Time-resolved and multi-state cryo-EM of TXNL1-, MIDN-, and ODC-bound proteasomes (preprints)","pmids":["bio_10.1101_2024.11.08.622731","bio_10.1101_2025.02.22.639686","bio_10.1101_2025.11.15.688597"],"confidence":"Medium","gaps":["Preprints not yet peer-reviewed","Functional consequences in cells not established"]},{"year":null,"claim":"It remains unresolved how PSMD14 is physically and functionally partitioned between its proteasome-bound deubiquitinase role and its numerous proteasome-independent activities on individual substrates and chromatin, and what governs substrate selection at the cellular level.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No systematic distinction between proteasome-associated and free-enzyme substrate processing","Recruitment determinants for specific substrates unknown","Quantitative cellular pools of proteasomal vs non-proteasomal PSMD14 not measured"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,2,5,6,13]},{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[0,6,16]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[34]},{"term_id":"GO:0140299","term_label":"molecular sensor activity","supporting_discovery_ids":[34]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[28,9]},{"term_id":"GO:0000228","term_label":"nuclear chromosome","supporting_discovery_ids":[9,28]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[24]}],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,2,5]},{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[9,10]},{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[28]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[24,27]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[24]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[8]}],"complexes":["19S proteasome regulatory particle (lid)","26S proteasome","PSMD14-NSD2 chromatin complex"],"partners":["PSMD7","PSMD10","NSD2","E2F1","SNAIL","TGFBR1","MYC","TXNL1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O00487","full_name":"Ubiquitin C-terminal hydrolase PSMD14","aliases":["26S proteasome non-ATPase regulatory subunit 14","26S proteasome regulatory subunit RPN11","26S proteasome-associated PAD1 homolog 1"],"length_aa":310,"mass_kda":34.6,"function":"Component of the 26S proteasome, a multiprotein complex involved in the ATP-dependent degradation of ubiquitinated proteins. This complex plays a key role in the maintenance of protein homeostasis by removing misfolded or damaged proteins, which could impair cellular functions, and by removing proteins whose functions are no longer required. Therefore, the proteasome participates in numerous cellular processes, including cell cycle progression, apoptosis, or DNA damage repair (PubMed:9374539, PubMed:1317798). The PSMD14 subunit is a metalloprotease that specifically cleaves 'Lys-63'-linked polyubiquitin chains within the complex (PubMed:22909820). Plays a role in response to double-strand breaks (DSBs): acts as a regulator of non-homologous end joining (NHEJ) by cleaving 'Lys-63'-linked polyubiquitin, thereby promoting retention of JMJD2A/KDM4A on chromatin and restricting TP53BP1 accumulation (PubMed:22909820). Also involved in homologous recombination repair by promoting RAD51 loading (PubMed:22909820). Regulates macroautophagy by ensuring Golgi-to-ER retrograde transport through its deubiquitinating activity on K63-linked ubiquitin chains. This activity prevents the retention of essential autophagy proteins at the Golgi, enabling their trafficking to autophagosome formation sites and supporting Golgi-ER membrane recycling critical for effective autophagy (PubMed:32210007)","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/O00487/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/PSMD14","classification":"Common Essential","n_dependent_lines":1205,"n_total_lines":1208,"dependency_fraction":0.9975165562913907},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"PSMD12","stoichiometry":10.0},{"gene":"PSMC4","stoichiometry":0.2},{"gene":"TOP1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/PSMD14","total_profiled":1310},"omim":[{"mim_id":"607274","title":"UBIQUITIN-SPECIFIC PROTEASE 14; USP14","url":"https://www.omim.org/entry/607274"},{"mim_id":"607173","title":"PROTEASOME 26S SUBUNIT, NON-ATPase, 14; PSMD14","url":"https://www.omim.org/entry/607173"},{"mim_id":"300617","title":"BRCA1/BRCA2-CONTAINING COMPLEX, SUBUNIT 3; BRCC3","url":"https://www.omim.org/entry/300617"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/PSMD14"},"hgnc":{"alias_symbol":["POH1","pad1","Rpn11"],"prev_symbol":[]},"alphafold":{"accession":"O00487","domains":[{"cath_id":"3.40.140.10","chopping":"29-219","consensus_level":"high","plddt":84.7158,"start":29,"end":219},{"cath_id":"1.10.287","chopping":"224-308","consensus_level":"high","plddt":87.4381,"start":224,"end":308}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O00487","model_url":"https://alphafold.ebi.ac.uk/files/AF-O00487-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O00487-F1-predicted_aligned_error_v6.png","plddt_mean":81.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=PSMD14","jax_strain_url":"https://www.jax.org/strain/search?query=PSMD14"},"sequence":{"accession":"O00487","fasta_url":"https://rest.uniprot.org/uniprotkb/O00487.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O00487/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O00487"}},"corpus_meta":[{"pmid":"12183636","id":"PMC_12183636","title":"Role of Rpn11 metalloprotease in deubiquitination and degradation by the 26S proteasome.","date":"2002","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/12183636","citation_count":852,"is_preprint":false},{"pmid":"19214193","id":"PMC_19214193","title":"K63-specific deubiquitination by two JAMM/MPN+ complexes: BRISC-associated Brcc36 and proteasomal Poh1.","date":"2009","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/19214193","citation_count":194,"is_preprint":false},{"pmid":"12370088","id":"PMC_12370088","title":"MPN+, a putative catalytic motif found in a subset of MPN domain proteins from eukaryotes and prokaryotes, is critical for Rpn11 function.","date":"2002","source":"BMC biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12370088","citation_count":181,"is_preprint":false},{"pmid":"22909820","id":"PMC_22909820","title":"The proteasomal de-ubiquitinating enzyme POH1 promotes the double-strand DNA break response.","date":"2012","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/22909820","citation_count":136,"is_preprint":false},{"pmid":"24463465","id":"PMC_24463465","title":"Structure of the Rpn11-Rpn8 dimer reveals mechanisms of substrate deubiquitination during proteasomal degradation.","date":"2014","source":"Nature structural & molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/24463465","citation_count":133,"is_preprint":false},{"pmid":"28244987","id":"PMC_28244987","title":"Capzimin is a potent and specific inhibitor of proteasome isopeptidase Rpn11.","date":"2017","source":"Nature chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/28244987","citation_count":129,"is_preprint":false},{"pmid":"14581483","id":"PMC_14581483","title":"Complementary roles for Rpn11 and Ubp6 in deubiquitination and proteolysis by the proteasome.","date":"2003","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/14581483","citation_count":121,"is_preprint":false},{"pmid":"24516147","id":"PMC_24516147","title":"Crystal structure of the proteasomal deubiquitylation module Rpn8-Rpn11.","date":"2014","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/24516147","citation_count":114,"is_preprint":false},{"pmid":"28459440","id":"PMC_28459440","title":"Thiolutin is a zinc chelator that inhibits the Rpn11 and other JAMM metalloproteases.","date":"2017","source":"Nature chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/28459440","citation_count":110,"is_preprint":false},{"pmid":"26510456","id":"PMC_26510456","title":"POH1 deubiquitylates and stabilizes E2F1 to promote tumour formation.","date":"2015","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/26510456","citation_count":98,"is_preprint":false},{"pmid":"24013561","id":"PMC_24013561","title":"Co-operation of BRCA1 and POH1 relieves the barriers posed by 53BP1 and RAP80 to resection.","date":"2013","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/24013561","citation_count":97,"is_preprint":false},{"pmid":"31634528","id":"PMC_31634528","title":"Deubiquitinase PSMD14 enhances hepatocellular carcinoma growth and metastasis by stabilizing GRB2.","date":"2019","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/31634528","citation_count":93,"is_preprint":false},{"pmid":"30590814","id":"PMC_30590814","title":"ECT2/PSMD14/PTTG1 axis promotes the proliferation of glioma through stabilizing E2F1.","date":"2019","source":"Neuro-oncology","url":"https://pubmed.ncbi.nlm.nih.gov/30590814","citation_count":92,"is_preprint":false},{"pmid":"28844860","id":"PMC_28844860","title":"An AAA Motor-Driven Mechanical Switch in Rpn11 Controls Deubiquitination at the 26S Proteasome.","date":"2017","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/28844860","citation_count":87,"is_preprint":false},{"pmid":"28581522","id":"PMC_28581522","title":"Blockade of deubiquitylating enzyme Rpn11 triggers apoptosis in multiple myeloma cells and overcomes bortezomib resistance.","date":"2017","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/28581522","citation_count":83,"is_preprint":false},{"pmid":"29331416","id":"PMC_29331416","title":"Deubiquitinating enzyme PSMD14 promotes tumor metastasis through stabilizing SNAIL in human esophageal squamous cell carcinoma.","date":"2018","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/29331416","citation_count":72,"is_preprint":false},{"pmid":"9763452","id":"PMC_9763452","title":"A mutation in a novel yeast proteasomal gene, RPN11/MPR1, produces a cell cycle arrest, overreplication of nuclear and mitochondrial DNA, and an altered mitochondrial morphology.","date":"1998","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/9763452","citation_count":68,"is_preprint":false},{"pmid":"34382324","id":"PMC_34382324","title":"Deubiquitinase PSMD14 promotes ovarian cancer progression by decreasing enzymatic activity of PKM2.","date":"2021","source":"Molecular oncology","url":"https://pubmed.ncbi.nlm.nih.gov/34382324","citation_count":60,"is_preprint":false},{"pmid":"17237285","id":"PMC_17237285","title":"The JAMM motif of human deubiquitinase Poh1 is essential for cell viability.","date":"2007","source":"Molecular cancer therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/17237285","citation_count":60,"is_preprint":false},{"pmid":"33456565","id":"PMC_33456565","title":"Blockade of deubiquitinating enzyme PSMD14 overcomes chemoresistance in head and neck squamous cell carcinoma by antagonizing E2F1/Akt/SOX2-mediated stemness.","date":"2021","source":"Theranostics","url":"https://pubmed.ncbi.nlm.nih.gov/33456565","citation_count":59,"is_preprint":false},{"pmid":"33897885","id":"PMC_33897885","title":"The PSMD14 inhibitor Thiolutin as a novel therapeutic approach for esophageal squamous cell carcinoma through facilitating SNAIL degradation.","date":"2021","source":"Theranostics","url":"https://pubmed.ncbi.nlm.nih.gov/33897885","citation_count":56,"is_preprint":false},{"pmid":"19732767","id":"PMC_19732767","title":"Knockdown of human deubiquitinase PSMD14 induces cell cycle arrest and senescence.","date":"2009","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/19732767","citation_count":54,"is_preprint":false},{"pmid":"15018611","id":"PMC_15018611","title":"Participation of the proteasomal lid subunit Rpn11 in mitochondrial morphology and function is mapped to a distinct C-terminal domain.","date":"2004","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/15018611","citation_count":50,"is_preprint":false},{"pmid":"31685442","id":"PMC_31685442","title":"The deubiquitinating enzyme PSMD14 facilitates tumor growth and chemoresistance through stabilizing the ALK2 receptor in the initiation of BMP6 signaling pathway.","date":"2019","source":"EBioMedicine","url":"https://pubmed.ncbi.nlm.nih.gov/31685442","citation_count":47,"is_preprint":false},{"pmid":"16569633","id":"PMC_16569633","title":"The 19 S proteasomal subunit POH1 contributes to the regulation of c-Jun ubiquitination, stability, and subcellular localization.","date":"2006","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/16569633","citation_count":41,"is_preprint":false},{"pmid":"19436748","id":"PMC_19436748","title":"Regulation of ErbB2 receptor status by the proteasomal DUB POH1.","date":"2009","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/19436748","citation_count":39,"is_preprint":false},{"pmid":"30146242","id":"PMC_30146242","title":"Epidithiodiketopiperazines Inhibit Protein Degradation by Targeting Proteasome Deubiquitinase Rpn11.","date":"2018","source":"Cell chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/30146242","citation_count":38,"is_preprint":false},{"pmid":"30745168","id":"PMC_30745168","title":"POH1 contributes to hyperactivation of TGF-β signaling and facilitates hepatocellular carcinoma metastasis through deubiquitinating TGF-β receptors and caveolin-1.","date":"2019","source":"EBioMedicine","url":"https://pubmed.ncbi.nlm.nih.gov/30745168","citation_count":38,"is_preprint":false},{"pmid":"30315153","id":"PMC_30315153","title":"POH1 deubiquitinates pro-interleukin-1β and restricts inflammasome activity.","date":"2018","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/30315153","citation_count":37,"is_preprint":false},{"pmid":"25389291","id":"PMC_25389291","title":"Disassembly of Lys11 and mixed linkage polyubiquitin conjugates provides insights into function of proteasomal deubiquitinases Rpn11 and Ubp6.","date":"2014","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/25389291","citation_count":37,"is_preprint":false},{"pmid":"18172023","id":"PMC_18172023","title":"Dissection of the carboxyl-terminal domain of the proteasomal subunit Rpn11 in maintenance of mitochondrial structure and function.","date":"2008","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/18172023","citation_count":35,"is_preprint":false},{"pmid":"40000916","id":"PMC_40000916","title":"Histone lactylation stimulated upregulation of PSMD14 alleviates neuron PANoptosis through deubiquitinating PKM2 to activate PINK1-mediated mitophagy after traumatic brain injury.","date":"2025","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/40000916","citation_count":34,"is_preprint":false},{"pmid":"28535005","id":"PMC_28535005","title":"RPN11 deubiquitinase promotes proliferation and migration of breast cancer cells.","date":"2017","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/28535005","citation_count":33,"is_preprint":false},{"pmid":"37935198","id":"PMC_37935198","title":"The proteasome component PSMD14 drives myelomagenesis through a histone deubiquitinase activity.","date":"2023","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/37935198","citation_count":32,"is_preprint":false},{"pmid":"29573636","id":"PMC_29573636","title":"POH1 Knockdown Induces Cancer Cell Apoptosis via p53 and Bim.","date":"2018","source":"Neoplasia (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/29573636","citation_count":32,"is_preprint":false},{"pmid":"38017133","id":"PMC_38017133","title":"PSMD14 stabilizes estrogen signaling and facilitates breast cancer progression via deubiquitinating ERα.","date":"2023","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/38017133","citation_count":30,"is_preprint":false},{"pmid":"20058232","id":"PMC_20058232","title":"The 19S proteasomal lid subunit POH1 enhances the transcriptional activation by Mitf in osteoclasts.","date":"2010","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20058232","citation_count":29,"is_preprint":false},{"pmid":"39146936","id":"PMC_39146936","title":"Amelioration of nonalcoholic fatty liver disease by inhibiting the deubiquitylating enzyme RPN11.","date":"2024","source":"Cell metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/39146936","citation_count":28,"is_preprint":false},{"pmid":"23936414","id":"PMC_23936414","title":"Integrity of the Saccharomyces cerevisiae Rpn11 protein is critical for formation of proteasome storage granules (PSG) and survival in stationary phase.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23936414","citation_count":26,"is_preprint":false},{"pmid":"36328147","id":"PMC_36328147","title":"Deubiquitylase PSMD14 inhibits autophagy to promote ovarian cancer progression via stabilization of LRPPRC.","date":"2022","source":"Biochimica et biophysica acta. Molecular basis of disease","url":"https://pubmed.ncbi.nlm.nih.gov/36328147","citation_count":24,"is_preprint":false},{"pmid":"30002364","id":"PMC_30002364","title":"Rpn11-mediated ubiquitin processing in an ancestral archaeal ubiquitination system.","date":"2018","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/30002364","citation_count":24,"is_preprint":false},{"pmid":"11943459","id":"PMC_11943459","title":"Mitochondrial effects of the pleiotropic proteasomal mutation mpr1/rpn11: uncoupling from cell cycle defects in extragenic revertants.","date":"2002","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/11943459","citation_count":24,"is_preprint":false},{"pmid":"35269789","id":"PMC_35269789","title":"PSMD14 Targeting Triggers Paraptosis in Breast Cancer Cells by Inducing Proteasome Inhibition and Ca2+ Imbalance.","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/35269789","citation_count":23,"is_preprint":false},{"pmid":"12553011","id":"PMC_12553011","title":"The essential 26S proteasome subunit Rpn11 confers multidrug resistance to mammalian cells.","date":"2002","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/12553011","citation_count":22,"is_preprint":false},{"pmid":"36241058","id":"PMC_36241058","title":"Novel insights into the non-canonical roles of PSMD14/POH1/Rpn11 in proteostasis and in the modulation of cancer progression.","date":"2022","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/36241058","citation_count":20,"is_preprint":false},{"pmid":"31212367","id":"PMC_31212367","title":"Targeting POH1 inhibits prostate cancer cell growth and enhances the suppressive efficacy of androgen deprivation and docetaxel.","date":"2019","source":"The Prostate","url":"https://pubmed.ncbi.nlm.nih.gov/31212367","citation_count":20,"is_preprint":false},{"pmid":"35405117","id":"PMC_35405117","title":"Deubiquitinating enzyme PSMD14 facilitates gastric carcinogenesis through stabilizing PTBP1.","date":"2022","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/35405117","citation_count":17,"is_preprint":false},{"pmid":"21619884","id":"PMC_21619884","title":"A proteasome assembly defect in rpn3 mutants is associated with Rpn11 instability and increased sensitivity to stress.","date":"2011","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/21619884","citation_count":17,"is_preprint":false},{"pmid":"25540361","id":"PMC_25540361","title":"The proteasomal Rpn11 metalloprotease suppresses tombusvirus RNA recombination and promotes viral replication via facilitating assembly of the viral replicase complex.","date":"2014","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/25540361","citation_count":17,"is_preprint":false},{"pmid":"33154524","id":"PMC_33154524","title":"Targeting PSMD14 inhibits melanoma growth through SMAD3 stabilization.","date":"2020","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/33154524","citation_count":16,"is_preprint":false},{"pmid":"32210007","id":"PMC_32210007","title":"The Proteasomal Deubiquitinating Enzyme PSMD14 Regulates Macroautophagy by Controlling Golgi-to-ER Retrograde Transport.","date":"2020","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/32210007","citation_count":15,"is_preprint":false},{"pmid":"38061486","id":"PMC_38061486","title":"POH1 induces Smad3 deubiquitination and promotes lung cancer metastasis.","date":"2023","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/38061486","citation_count":14,"is_preprint":false},{"pmid":"30038387","id":"PMC_30038387","title":"Requirement for POH1 in differentiation and maintenance of regulatory T cells.","date":"2018","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/30038387","citation_count":14,"is_preprint":false},{"pmid":"30356695","id":"PMC_30356695","title":"Computational Studies on the Inhibitor Selectivity of Human JAMM Deubiquitinylases Rpn11 and CSN5.","date":"2018","source":"Frontiers in chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/30356695","citation_count":14,"is_preprint":false},{"pmid":"40444470","id":"PMC_40444470","title":"PSMD14 Stabilizes SLC7A11 to Ameliorate Glucocorticoid-Induced Osteoporosis by Suppressing Osteocyte Ferroptosis.","date":"2025","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/40444470","citation_count":12,"is_preprint":false},{"pmid":"34161398","id":"PMC_34161398","title":"A novel viral strategy for host factor recruitment: The co-opted proteasomal Rpn11 protein interaction hub in cooperation with subverted actin filaments are targeted to deliver cytosolic host factors for viral replication.","date":"2021","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/34161398","citation_count":11,"is_preprint":false},{"pmid":"20941496","id":"PMC_20941496","title":"Synthetic lethality of rpn11-1 rpn10Δ is linked to altered proteasome assembly and activity.","date":"2010","source":"Current genetics","url":"https://pubmed.ncbi.nlm.nih.gov/20941496","citation_count":11,"is_preprint":false},{"pmid":"40344056","id":"PMC_40344056","title":"Targeting PSMD14 combined with arachidonic acid induces synthetic lethality via FADS1 m6A modification in triple-negative breast cancer.","date":"2025","source":"Science advances","url":"https://pubmed.ncbi.nlm.nih.gov/40344056","citation_count":10,"is_preprint":false},{"pmid":"36632137","id":"PMC_36632137","title":"Depletion of PSMD14 suppresses bladder cancer proliferation by regulating GPX4.","date":"2023","source":"PeerJ","url":"https://pubmed.ncbi.nlm.nih.gov/36632137","citation_count":10,"is_preprint":false},{"pmid":"35328679","id":"PMC_35328679","title":"Proteomic Assessment of Extracellular Vesicles from Canine Tissue Explants as a Pipeline to Identify Molecular Targets in Osteosarcoma: PSMD14/Rpn11 as a Proof of Principle.","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/35328679","citation_count":10,"is_preprint":false},{"pmid":"40025166","id":"PMC_40025166","title":"ALKBH1 knockdown promotes the growth, migration and invasion of HTR-8/SVneo cells through regulating the m5C modification PSMD14.","date":"2025","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/40025166","citation_count":10,"is_preprint":false},{"pmid":"38053170","id":"PMC_38053170","title":"The function and mechanism of PSMD14 in promoting progression and resistance to anlotinib in osteosarcoma.","date":"2023","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/38053170","citation_count":8,"is_preprint":false},{"pmid":"40411784","id":"PMC_40411784","title":"The deubiquitinase Rpn11 functions as an allosteric ubiquitin sensor to promote substrate engagement by the 26S proteasome.","date":"2025","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/40411784","citation_count":8,"is_preprint":false},{"pmid":"38696072","id":"PMC_38696072","title":"Deubiquitinase PSMD14 promotes tumorigenicity of glioblastoma by deubiquitinating and stabilizing β-catenin.","date":"2024","source":"BioFactors (Oxford, England)","url":"https://pubmed.ncbi.nlm.nih.gov/38696072","citation_count":7,"is_preprint":false},{"pmid":"37844756","id":"PMC_37844756","title":"POH1 facilitates pancreatic carcinogenesis through MYC-driven acinar-to-ductal metaplasia and is a potential therapeutic target.","date":"2023","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/37844756","citation_count":7,"is_preprint":false},{"pmid":"40016178","id":"PMC_40016178","title":"PSMD14-mediated deubiquitination of CARM1 facilitates the proliferation and metastasis of hepatocellular carcinoma by inducing the transcriptional activation of FERMT1.","date":"2025","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/40016178","citation_count":7,"is_preprint":false},{"pmid":"38643468","id":"PMC_38643468","title":"An LRPPRC-HAPSTR1-PSMD14 interaction regulates tumor progression in ovarian cancer.","date":"2024","source":"Aging","url":"https://pubmed.ncbi.nlm.nih.gov/38643468","citation_count":7,"is_preprint":false},{"pmid":"21059189","id":"PMC_21059189","title":"Analysis of the rpn11-m1 proteasomal mutant reveals connection between cell cycle and mitochondrial biogenesis.","date":"2010","source":"FEMS yeast research","url":"https://pubmed.ncbi.nlm.nih.gov/21059189","citation_count":6,"is_preprint":false},{"pmid":"40066751","id":"PMC_40066751","title":"PSMD14 Transcriptionally Activated by MEF2A Promotes Pancreatic Cancer Development by Upregulating SPON2 Expression.","date":"2025","source":"The Kaohsiung journal of medical sciences","url":"https://pubmed.ncbi.nlm.nih.gov/40066751","citation_count":5,"is_preprint":false},{"pmid":"41051446","id":"PMC_41051446","title":"PSMD14-Mediated LDHA Deubiquitination Upregulates ACLY Expression via H3K18 Lactylation to Promote Lipid Synthesis and Pancreatic Cancer Progression.","date":"2025","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/41051446","citation_count":4,"is_preprint":false},{"pmid":"40902741","id":"PMC_40902741","title":"PSMD14 promotes breast cancer progression by reducing K63-linked ubiquitination on FOXM1 and activating the PI3K/AKT/mTOR pathway.","date":"2025","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/40902741","citation_count":4,"is_preprint":false},{"pmid":"40365861","id":"PMC_40365861","title":"PSMD14/E2F1 Axis-Mediated CENPF Promotes the Metastasis of Triple-Negative Breast Cancer Through Inhibiting Ferroptosis.","date":"2025","source":"Cancer science","url":"https://pubmed.ncbi.nlm.nih.gov/40365861","citation_count":4,"is_preprint":false},{"pmid":"30326263","id":"PMC_30326263","title":"New perspectives on Mega plasmid sequence (poh1) in Bacillus thuringiensis ATCC 10792 harbouring antimicrobial, insecticidal and antibiotic resistance genes.","date":"2018","source":"Microbial pathogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/30326263","citation_count":4,"is_preprint":false},{"pmid":"33113963","id":"PMC_33113963","title":"Transient Breakage of the Nucleocytoplasmic Barrier Controls Spore Maturation via Mobilizing the Proteasome Subunit Rpn11 in the Fission Yeast Schizosaccharomyces pombe.","date":"2020","source":"Journal of fungi (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/33113963","citation_count":4,"is_preprint":false},{"pmid":"36350544","id":"PMC_36350544","title":"High-Throughput Assay for Characterizing Rpn11 Deubiquitinase Activity.","date":"2023","source":"Methods in molecular biology (Clifton, N.J.)","url":"https://pubmed.ncbi.nlm.nih.gov/36350544","citation_count":3,"is_preprint":false},{"pmid":"37975230","id":"PMC_37975230","title":"Regulation of Drp1 and enhancement of mitochondrial fission by the deubiquitinating enzyme PSMD14 facilitates the proliferation of bladder cancer cells.","date":"2023","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/37975230","citation_count":3,"is_preprint":false},{"pmid":"40311241","id":"PMC_40311241","title":"Design and synthesis of thiolutin derived PSMD14/HDAC dual-target inhibitors against esophageal squamous cell carcinoma.","date":"2025","source":"Bioorganic chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/40311241","citation_count":3,"is_preprint":false},{"pmid":"41193870","id":"PMC_41193870","title":"PSMD14-mediated PFKFB2 deubiquitination activates H3K27 lactylation to drive cancer stemness in gastric adenocarcinoma.","date":"2025","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/41193870","citation_count":2,"is_preprint":false},{"pmid":"34572530","id":"PMC_34572530","title":"An Arsenite Relay between PSMD14 and AIRAP Enables Revival of Proteasomal DUB Activity.","date":"2021","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/34572530","citation_count":2,"is_preprint":false},{"pmid":"40262717","id":"PMC_40262717","title":"Blocking PSMD14-mediated E2F1/ERK/AKT signaling pathways suppresses the progression of anaplastic thyroid cancer.","date":"2025","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/40262717","citation_count":1,"is_preprint":false},{"pmid":"41488674","id":"PMC_41488674","title":"PSMD14 drives lung adenocarcinoma progression through HMMR stabilization and dual activation of TGF-β/Smad and PI3K/AKT/mTOR signaling.","date":"2025","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/41488674","citation_count":1,"is_preprint":false},{"pmid":"40833626","id":"PMC_40833626","title":"MY-1 promotes angiogenesis in the ischemic hindlimbs by regulating the stability of CDC42 via PSMD14.","date":"2025","source":"Angiogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/40833626","citation_count":1,"is_preprint":false},{"pmid":"25509418","id":"PMC_25509418","title":"[Design of cell line stable expressing proteasomal subunit PSMD14 fused to the fluorescent protein EGFP and HTBH tag based on HEK293 cells].","date":"2014","source":"Tsitologiia","url":"https://pubmed.ncbi.nlm.nih.gov/25509418","citation_count":1,"is_preprint":false},{"pmid":"41876842","id":"PMC_41876842","title":"Targeting the PSMD14-BCKDK pathway overcomes immune suppression and enhances CAR-NK infiltration in glioblastoma.","date":"2026","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/41876842","citation_count":0,"is_preprint":false},{"pmid":"41608571","id":"PMC_41608571","title":"Coupling proteostasis and de novo purine biosynthesis of PSMD14 fuels glioblastoma progression and chemoresistance.","date":"2026","source":"Theranostics","url":"https://pubmed.ncbi.nlm.nih.gov/41608571","citation_count":0,"is_preprint":false},{"pmid":"41981629","id":"PMC_41981629","title":"Targeting PSMD14 enhances immunotherapy efficacy by promoting PD-L1 degradation and reshaping the tumor microenvironment in breast cancer.","date":"2026","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/41981629","citation_count":0,"is_preprint":false},{"pmid":"42174657","id":"PMC_42174657","title":"DUSP4-mediated dephosphorylation of PSMD14 enhances PSMD14-dependent deubiquitination of MICALL2 and promotes malignant progression in clear cell renal cell carcinoma.","date":"2026","source":"Journal of translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/42174657","citation_count":0,"is_preprint":false},{"pmid":"39484543","id":"PMC_39484543","title":"The deubiquitinase Rpn11 functions as an allosteric ubiquitin sensor to promote substrate engagement by the 26S proteasome.","date":"2024","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/39484543","citation_count":0,"is_preprint":false},{"pmid":"41647277","id":"PMC_41647277","title":"Covalent targeting of PSMD14 by Eupalinolide B induces oncoprotein degradation and apoptosis in acute promyelocytic leukemia cells.","date":"2026","source":"RSC chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/41647277","citation_count":0,"is_preprint":false},{"pmid":"41650523","id":"PMC_41650523","title":"Ethyl ferulate suppresses choroidal neovascularization by accelerating Keap1 degradation through the inhibition of PSMD14-mediated deubiquitination.","date":"2026","source":"Phytomedicine : international journal of phytotherapy and phytopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/41650523","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.08.13.670075","title":"USP15: the fourth Proteasome-associated DUB","date":"2025-08-13","source":"bioRxiv","url":"https://doi.org/10.1101/2025.08.13.670075","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.02.22.639686","title":"Structural basis for the midnolin-proteasome pathway and its role in suppressing myeloma","date":"2025-02-23","source":"bioRxiv","url":"https://doi.org/10.1101/2025.02.22.639686","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.11.08.622741","title":"Structure of the TXNL1-bound proteasome","date":"2024-11-10","source":"bioRxiv","url":"https://doi.org/10.1101/2024.11.08.622741","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.07.31.667872","title":"Structures of dynamic interactors at native proteasomes by PhIX-MS and cryoelectron microscopy","date":"2025-08-02","source":"bioRxiv","url":"https://doi.org/10.1101/2025.07.31.667872","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.11.15.688597","title":"Mechanisms of Ub-independent ODC recognition and translocation by the 26S proteasome revealing proteasome’s functional adaptability","date":"2025-11-16","source":"bioRxiv","url":"https://doi.org/10.1101/2025.11.15.688597","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.11.08.622731","title":"Structural landscape of AAA+ ATPase motor states in the substrate-degrading human 26S proteasome reveals conformation-specific binding of TXNL1","date":"2024-11-09","source":"bioRxiv","url":"https://doi.org/10.1101/2024.11.08.622731","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.03.04.641557","title":"Structural insights into the ubiquitin independent MIDN-proteasome pathway","date":"2025-03-11","source":"bioRxiv","url":"https://doi.org/10.1101/2025.03.04.641557","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.12.05.627083","title":"Insights into Dentatorubral-Pallidoluysian Atrophy from a new  <i>Drosophila</i>  model of disease","date":"2024-12-10","source":"bioRxiv","url":"https://doi.org/10.1101/2024.12.05.627083","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.09.18.613619","title":"The binary protein interactome mapping of the  <i>Giardia lamblia</i>  proteasome lid reveals extra proteasomal functions of GlRpn11","date":"2024-09-19","source":"bioRxiv","url":"https://doi.org/10.1101/2024.09.18.613619","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":50324,"output_tokens":13401,"usd":0.175993,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":25293,"output_tokens":5766,"usd":0.135307,"stage2_stop_reason":"end_turn"},"total_usd":0.3113,"stage1_batch_id":"msgbatch_01CNoDbkChHEkgrJnL7BwtGJ","stage2_batch_id":"msgbatch_016Y4x9E9Wg5VQDANj9quSQD","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2002,\n      \"finding\": \"Rpn11 is a metalloprotease deubiquitinase (DUB) residing in the proteasome lid subcomplex. Its JAMM motif (EX(n)HXHX(10)D) constitutes the active site; mutation of the predicted catalytic histidines to alanine (rpn11AXA) was lethal in yeast and caused mutant proteasomes—which assembled normally—to fail to deubiquitinate or degrade ubiquitinated Sic1 in vitro, demonstrating that Rpn11-mediated deubiquitination is coupled to substrate degradation.\",\n      \"method\": \"Active-site mutagenesis (rpn11AXA), yeast genetics (lethality), in vitro reconstituted degradation assay with purified mutant proteasomes\",\n      \"journal\": \"Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with mutagenesis, functional coupling demonstrated, independently replicated across multiple subsequent studies\",\n      \"pmids\": [\"12183636\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"The MPN+ motif (five conserved polar residues resembling metalloprotease active-site residues) is essential for Rpn11 function. Single amino acid substitutions in MPN+ residues cause slow growth, temperature sensitivity, and proteasome-dependent proteolysis defects in yeast, while a conserved Cys outside MPN+ is not essential.\",\n      \"method\": \"Site-directed mutagenesis of MPN+ motif residues, yeast phenotypic analysis, proteasome proteolysis assays\",\n      \"journal\": \"BMC Biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — mutagenesis with multiple mutant alleles, consistent phenotypes, replicated findings\",\n      \"pmids\": [\"12370088\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"Rpn11 and Ubp6 serve complementary roles in proteasomal deubiquitination. Proteasomes purified from rpn11 catalytic-motif mutants or ubp6 null strains both show slower deubiquitination rates; the double mutant is synthetically lethal. Rpn11-containing proteasomes show sensitivity to metal chelators consistent with Rpn11 being a metalloprotein. Degradation requires the intact lid (lidless proteasomes deubiquitinate but do not degrade ubiquitinated protein).\",\n      \"method\": \"Purification of proteasomes from yeast mutants, in vitro deubiquitination assays, metal chelator treatment, synthetic lethality genetics\",\n      \"journal\": \"Journal of Biological Chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal biochemical approaches, genetic epistasis, replicated in subsequent studies\",\n      \"pmids\": [\"14581483\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Rpn11 has two separable functional domains: its N-terminal MPN+/JAMM catalytic domain mediates proteasome-associated deubiquitination required for proteolysis, while a distinct C-terminal domain is required for mitochondrial morphology maintenance and is independent of catalytic activity. Overexpression of wild-type Rpn8 rescues cell cycle but not mitochondrial defects of rpn11 C-terminal mutant (mpr1-1). RPN8-RPN11 chimera analysis confirmed the C-terminal region of Rpn11 is necessary and sufficient to rescue mitochondrial phenotypes.\",\n      \"method\": \"Yeast genetics, RPN8-RPN11 chimera expression, intragenic complementation, mitochondrial morphology microscopy, cell cycle analysis\",\n      \"journal\": \"Biochemical Journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple chimeric constructs, complementation assays, functional dissection of two independent domains replicated across multiple studies\",\n      \"pmids\": [\"15018611\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"POH1/PSMD14 deubiquitinates c-Jun in mammalian cells. Ectopic POH1 expression in HEK293 cells decreased c-Jun ubiquitination, stabilized c-Jun protein, redistributed it to the nucleus, and increased AP1-mediated gene expression. Mutation of Cys-120 in the MPN+ motif reduced these effects. The stabilization appeared selective for c-Jun among tested proteasomal substrates.\",\n      \"method\": \"Ectopic overexpression, ubiquitination assays, reporter assays for AP1 transcription, active-site mutagenesis (C120 mutation)\",\n      \"journal\": \"Journal of Biological Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single lab, Co-IP/ubiquitination assay plus functional reporter, selectivity claim based on limited substrate panel\",\n      \"pmids\": [\"16569633\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"The JAMM zinc metalloproteinase motif of human POH1/PSMD14 is essential for cell viability and 26S proteasome function. RNAi knockdown of endogenous POH1 reduced viability and elevated polyubiquitinated proteins; rescue with wild-type POH1 restored viability, but a JAMM active-site double histidine-to-alanine mutant failed to rescue, confirming catalytic activity is required.\",\n      \"method\": \"siRNA knockdown, RNAi complementation with wild-type vs. JAMM-mutant POH1, cell viability assays, polyubiquitin accumulation analysis in HeLa cells\",\n      \"journal\": \"Molecular Cancer Therapeutics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — RNAi complementation with active-site mutant, functional rescue assay, single lab but orthogonal methods\",\n      \"pmids\": [\"17237285\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"POH1/PSMD14 within the PA700/19S proteasome is a K63-specific deubiquitinase. Biochemical fractionation of HeLa cell extracts through seven chromatographic steps co-purified the K63-specific DUB activity with the 19S proteasome. This activity was intrinsic to PA700 and was insensitive to N-ethylmaleimide and ubiquitin aldehyde (ruling out cysteine-based DUBs), consistent with JAMM metalloprotease activity. None of the complexes cleaved K6, K11, K29, K48, or alpha-linked chains.\",\n      \"method\": \"Multi-step chromatographic co-fractionation, linkage-specific deubiquitination assays, inhibitor profiling (NEM, ubiquitin aldehyde), gel filtration\",\n      \"journal\": \"EMBO Journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — seven-step biochemical co-purification plus multiple linkage-specificity assays, single lab but rigorous biochemistry\",\n      \"pmids\": [\"19214193\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"POH1/PSMD14 knockdown causes apparent loss of ErbB2 protein in HeLa cells, explained at least partially by accumulation of higher-molecular-weight ubiquitinated forms of ErbB2 rather than increased degradation rate. POH1 appears to deubiquitinate ErbB2 in a manner not necessarily coupled to proteasomal degradation. Cell-surface ErbB2 levels were only mildly affected.\",\n      \"method\": \"siRNA library screen for DUBs, Western blot, flow cytometry for surface ErbB2, comparison with proteasome inhibitor epoxomicin\",\n      \"journal\": \"PLoS ONE\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single Co-IP/knockdown approach, single lab, partial mechanistic follow-up\",\n      \"pmids\": [\"19436748\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"PSMD14 knockdown in carcinoma cell lines causes G0/G1 cell cycle arrest and cellular senescence, associated with downregulation of cyclin B1-CDK1-CDC25C and cyclin D1, upregulation of p21 and p27, and markedly reduced retinoblastoma protein phosphorylation. These effects are distinct from those caused by knockdown of PSMB5 (20S subunit), indicating that the 19S and 20S proteasome subunits have distinct biological functions.\",\n      \"method\": \"siRNA knockdown, flow cytometry (cell cycle), Western blot for cell cycle regulators, comparative siRNA of PSMB5\",\n      \"journal\": \"Experimental Cell Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KD with specific phenotypic readouts, comparison with 20S component, single lab\",\n      \"pmids\": [\"19732767\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"POH1/PSMD14 processes K63-linked polyubiquitin chains generated at DNA double-strand break (DSB) sites, thereby limiting 53BP1 accumulation (via antagonism of RNF8/RNF168-mediated K63-Ub) and promoting JMJD2A chromatin retention. POH1 also promotes RAD51 loading in homologous recombination independently of 53BP1. POH1-deficient cells show increased sensitivity to DNA damaging agents.\",\n      \"method\": \"siRNA knockdown, immunofluorescence for DSB foci (53BP1, RAD51, γH2AX), K63-Ub chromatin analysis, cell survival assays\",\n      \"journal\": \"EMBO Journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods, multiple pathway endpoints, functionally linked to DNA repair mechanism\",\n      \"pmids\": [\"22909820\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"POH1 relieves the barrier posed by RAP80 to DNA end resection in G2 phase. POH1 depletion enlarges 53BP1 and ubiquitin chain foci and prevents formation of an RPA-positive resection core. Co-depletion of POH1 with RAP80, BRCC36, or ABRAXAS restores the resection-competent core, suggesting POH1 removes ubiquitin chains in the IRIF core that RAP80 reads, enabling transition from NHEJ to HR. BRCA1 and POH1 act as distinct but interfacing barriers to ubiquitin chain removal.\",\n      \"method\": \"siRNA co-depletion experiments, immunofluorescence for DSB factors (53BP1, RAP80, RPA, ubiquitin FK2), cell cycle-staged analysis\",\n      \"journal\": \"Nucleic Acids Research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple co-depletion epistasis experiments, specific molecular readouts, pathway position defined\",\n      \"pmids\": [\"24013561\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Crystal structures of the Rpn11-Rpn8 heterodimer at 2.0 Å resolution reveal that Rpn11 lacks a conserved surface for ubiquitin Ile44-patch binding, does not contact the proximal side of the scissile isopeptide bond, and exhibits no ubiquitin linkage specificity. Two distinct interfaces mediate the Rpn11-Rpn8 interaction. Mutational studies confirm these structural features explain how Rpn11 functions as a promiscuous, cotranslocational deubiquitinase.\",\n      \"method\": \"X-ray crystallography (Zn2+-free and Zn2+-bound structures), site-directed mutagenesis, biochemical DUB assays\",\n      \"journal\": \"Nature Structural & Molecular Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure + mutagenesis + functional validation, replicated by independent structure paper (PMID:24516147)\",\n      \"pmids\": [\"24463465\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Crystal structures of the Rpn8-Rpn11 MPN-domain heterodimer (obtained via nanobody-assisted crystallization) reveal that full Rpn11 activation requires incorporation into the 26S proteasome and is dependent on ATP hydrolysis. Premature activation is prevented by: (1) low intrinsic ubiquitin affinity, (2) an insertion segment acting as a physical barrier across the substrate access channel, and (3) a conformationally unstable catalytic loop. Docking into proteasome EM density shows Rpn11 contacts ATPase subunits that stabilize the active conformation.\",\n      \"method\": \"X-ray crystallography (three structures), cryo-EM density docking, nanobody-assisted crystallization, DUB activity assay with model substrate\",\n      \"journal\": \"PNAS\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure with functional validation, complementary to PMID:24463465\",\n      \"pmids\": [\"24516147\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Rpn11 and Ubp6 process K11- and K63-linked ubiquitin chains with comparable efficiencies (increasing with chain length), whereas proteasomal processing of K48-linked chains is inversely correlated with chain length. Incorporation into proteasomes enhances Rpn11 enzymatic efficiency by roughly 2 orders of magnitude, partly by relieving autoinhibition by its C-terminus. Rpn11 shows a random cleavage mode on K48 chains (broad/endo), while Ubp6 shows endo-chain preference on K48.\",\n      \"method\": \"In vitro DUB assays with defined ubiquitin conjugates (homogeneous K11, K48, K63 chains of varying lengths, heterogeneous chains), fluorescently labeled Ub chains, purified proteasomes vs isolated DUBs\",\n      \"journal\": \"Journal of Biological Chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — rigorous in vitro biochemistry with panel of defined substrates, single lab but multiple orthogonal assays\",\n      \"pmids\": [\"25389291\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"POH1/PSMD14 deubiquitylates and stabilizes E2F1. POH1 physically binds E2F1 and removes ubiquitin from it, preventing its proteasomal degradation. Conditional knockout of Poh1 in primary mouse liver cells reduces E2F1 protein levels. Stabilized E2F1 upregulates Survivin and FOXM1, promoting liver cancer cell tumor growth.\",\n      \"method\": \"Co-immunoprecipitation, in vivo ubiquitination assays, conditional Poh1 knockout mouse model, Western blot, xenograft mouse model\",\n      \"journal\": \"Nature Communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, in vivo ubiquitination assay, conditional KO mouse, replicated in multiple cancer-type papers\",\n      \"pmids\": [\"26510456\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Rpn11 DUB activity is coupled to substrate translocation by the AAA+ ATPase motor via a conformational switch of the Insert-1 (Ins-1) loop. Ubiquitin binding induces Ins-1 transition from an inactive closed state to an active β-hairpin; this switch is rate-limiting for deubiquitination and is strongly accelerated by mechanical substrate translocation. Deubiquitination by Rpn11 and ubiquitin unfolding by the ATPases are in direct competition, requiring rapid Rpn11 activation to prevent ubiquitin co-degradation.\",\n      \"method\": \"X-ray crystallography (ubiquitin-bound Rpn11 structure), mutagenesis, in vitro single-molecule and ensemble DUB assays, biochemical translocation-deubiquitination coupling assays\",\n      \"journal\": \"Molecular Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure of ubiquitin-bound state, mutagenesis, multiple biochemical assays establishing mechanochemical coupling\",\n      \"pmids\": [\"28844860\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Quinoline-8-thiol derivative capzimin is a selective inhibitor of proteasomal Rpn11/PSMD14 that inhibits the JAMM metalloprotease by chelating its active-site zinc. Capzimin (>5-fold selectivity for Rpn11 over related JAMM proteases) stabilizes polyubiquitinated proteasome substrates, induces unfolded protein response, and blocks cancer cell proliferation including bortezomib-resistant cells.\",\n      \"method\": \"Biochemical DUB activity assays, proteomic analysis of stabilized substrates, cell viability assays, selectivity profiling against related JAMM proteases and metalloenzymes\",\n      \"journal\": \"Nature Chemical Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — biochemical characterization, selectivity profiling, cellular proteomics, multiple cell line validation\",\n      \"pmids\": [\"28244987\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Thiolutin, a disulfide-containing antibiotic, is a zinc chelator that inhibits the JAMM metalloprotease Rpn11/PSMD14 of the 19S proteasome in its reduced form. It also inhibits related JAMM metalloproteases Csn5, AMSH, and BRCC36.\",\n      \"method\": \"Biochemical DUB inhibition assays, zinc chelation characterization, zinc-binding experiments with Rpn11 and related JAMM enzymes\",\n      \"journal\": \"Nature Chemical Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro biochemical inhibition assays across multiple JAMM family members, zinc chelation mechanism established\",\n      \"pmids\": [\"28459440\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"PSMD14 deubiquitylates SNAIL (EMT transcription factor), preventing its ubiquitin-mediated proteasomal degradation, thereby stabilizing SNAIL protein. Mass spectrometry identified PSMD14 as a SNAIL-interacting DUB. PSMD14 knockdown blocks SNAIL-induced EMT, suppresses tumor cell migration and invasion in vitro, and inhibits metastasis in vivo.\",\n      \"method\": \"Mass spectrometry interactome screen, co-immunoprecipitation, in vivo ubiquitination assay, siRNA knockdown with migration/invasion assays, xenograft metastasis model\",\n      \"journal\": \"Cancer Letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MS-identified interaction, reciprocal Co-IP, in vivo ubiquitination, functional in vivo validation\",\n      \"pmids\": [\"29331416\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Epidithiodiketopiperazines (ETPs) inhibit proteasomal degradation by targeting Rpn11/POH1/PSMD14, the essential proteasomal DUB. ETPs also inhibit related JAMM proteases Csn5 and AMSH. An improved ETP (SOP11) stabilizes a subset of polyubiquitinated proteasome substrates, induces the unfolded protein response, and causes cell death.\",\n      \"method\": \"In vitro reconstituted proteasome-mediated protein degradation assay, biochemical Rpn11 inhibition assays, cellular proteasome substrate stabilization, UPR reporter assays\",\n      \"journal\": \"Cell Chemical Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstituted degradation assay plus cellular validation, single lab\",\n      \"pmids\": [\"30146242\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"POH1/PSMD14 deubiquitinates pro-IL-1β by removing K63-linked polyubiquitin chains, decreasing its susceptibility to cleavage and mature IL-1β production. POH1 physically interacts with pro-IL-1β. Myeloid cell-specific POH1 deletion aggravates LPS-induced systemic inflammation and alum-induced peritonitis in vivo.\",\n      \"method\": \"Co-immunoprecipitation, K63-specific deubiquitination assay, myeloid-specific conditional knockout mice, in vivo inflammation models (LPS, alum peritonitis), IL-1β production measurement\",\n      \"journal\": \"Nature Communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — Co-IP, linkage-specific ubiquitination assay, conditional KO mouse with in vivo phenotype\",\n      \"pmids\": [\"30315153\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"PSMD14 deubiquitinates and stabilizes GRB2 via inhibiting its ubiquitin-mediated proteasomal degradation, promoting HCC proliferation, migration, and invasion.\",\n      \"method\": \"Co-immunoprecipitation, in vivo ubiquitination assay, siRNA knockdown, overexpression experiments, xenograft and metastasis mouse models\",\n      \"journal\": \"Cancer Letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and ubiquitination assay, in vivo validation, single lab\",\n      \"pmids\": [\"31634528\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"PSMD14 deubiquitinates the ALK2 (BMP type I receptor) by removing K48-linked ubiquitin chains added by Smurf1 E3 ligase, thereby stabilizing ALK2 and promoting BMP6 signaling pathway activation. This function is stated to be independent of its intrinsic role in the 26S proteasome. PSMD14 was identified via siRNA DUB library screen.\",\n      \"method\": \"siRNA DUB library screen, immunoblot, co-immunoprecipitation, K48-specific ubiquitination assays, xenograft colorectal cancer model\",\n      \"journal\": \"EBioMedicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — DUB screen, Co-IP, linkage-specific ubiquitination assay, in vivo validation, single lab\",\n      \"pmids\": [\"31685442\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"POH1 deubiquitinates TGF-β receptors (TGFBR1 and TGFBR2) and caveolin-1 (CAV1), preventing lysosome pathway-mediated turnover of TGF-β receptors and thereby hyperactivating TGF-β signaling. POH1-deficient mouse hepatocytes show severely downregulated TGF-β receptor levels. This promotes HCC metastatic properties in vitro and in vivo.\",\n      \"method\": \"Western blotting, co-immunoprecipitation, ubiquitination assays, conditional Poh1 knockout mouse (Mx-Cre+, poh1f/f), xenograft and metastasis models\",\n      \"journal\": \"EBioMedicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — Co-IP, ubiquitination assay, conditional KO mouse with in vivo phenotype, multiple substrates investigated\",\n      \"pmids\": [\"30745168\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"PSMD14 depletion or pharmacological inhibition (capzimin) causes retention of Atg9A and Rab1A at the Golgi apparatus, blocking Golgi-to-ER retrograde transport and consequently inhibiting macroautophagy. PSMD14 acts specifically on K63-linked ubiquitin chains in cells. Inhibition of the 20S proteasome did not recapitulate these trafficking effects, indicating a specific role for PSMD14/K63-Ub in Golgi-to-ER retrograde transport.\",\n      \"method\": \"High-content siRNA screening (1187 ubiquitinome genes), APP trafficking reporter, siRNA knockdown, capzimin treatment, fluorescence microscopy for Golgi/ER trafficking markers, autophagy flux assays\",\n      \"journal\": \"Cells\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genome-scale siRNA screen, pharmacological validation, specific trafficking assays, 20S vs 19S comparison as control\",\n      \"pmids\": [\"32210007\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Arsenite inhibits PSMD14/Rpn11 metalloprotease DUB activity by substituting zinc in the MPN/JAMM domain. The proteasomal adaptor AIRAP can directly relieve this PSMD14/Rpn11 inhibition, suggesting a metal relay mechanism between arsenylated PSMD14 and AIRAP to restore proteasomal DUB function during arsenite stress.\",\n      \"method\": \"Direct arsenite binding assay, in vitro Rpn11 DUB inhibition assay, AIRAP interaction and relief-of-inhibition experiments\",\n      \"journal\": \"Biomolecules\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — biochemical inhibition and relief assays, single lab, mechanistic model proposed but limited orthogonal validation\",\n      \"pmids\": [\"34572530\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"PSMD14 decreases K63-linked ubiquitination on PKM2, shifts the PKM2 oligomeric equilibrium from tetramers toward dimers/monomers, diminishes pyruvate kinase enzymatic activity, and induces nuclear translocation of PKM2, thereby promoting aerobic glycolysis in ovarian cancer cells.\",\n      \"method\": \"Co-immunoprecipitation, K63-specific ubiquitination assays, PKM2 oligomeric state analysis (native PAGE), pyruvate kinase activity assay, nuclear fractionation, siRNA knockdown/overexpression\",\n      \"journal\": \"Molecular Oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — linkage-specific ubiquitination, enzymatic activity assay, fractionation, single lab\",\n      \"pmids\": [\"34382324\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"PSMD14 deubiquitinates and stabilizes LRPPRC by inhibiting its ubiquitination, thereby suppressing autophagy through the LRPPRC/Beclin1-Bcl-2/SQSTM1 signaling pathway in ovarian cancer cells.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays, autophagy flux assays, siRNA knockdown, xenograft and metastasis mouse models\",\n      \"journal\": \"BBA Molecular Basis of Disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assay, pathway analysis, in vivo validation, single lab\",\n      \"pmids\": [\"36328147\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"PSMD14 acts as a histone H2AK119 deubiquitinase independently of the 19S regulatory particle, functioning on chromatin in complex with NSD2. This non-proteasomal PSMD14 activity facilitates NSD2-directed H3K36 dimethylation and transcriptional activation of target genes including RELA, driving myelomagenesis. RELA in turn transactivates PSMD14, forming a positive feedback loop.\",\n      \"method\": \"ChIP-seq, co-immunoprecipitation (chromatin-bound PSMD14-NSD2 complex), histone H2AK119 deubiquitination assays, integrative genomic/epigenomic analyses, PSMD14 inhibitor studies\",\n      \"journal\": \"Molecular Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — chromatin biochemistry, histone DUB assay, integrative epigenomics, functional feedback loop demonstrated\",\n      \"pmids\": [\"37935198\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"POH1 deubiquitinates and stabilizes the MYC protein, which potentiates acinar-to-ductal metaplasia (ADM) and pancreatic ductal adenocarcinoma (PDAC). Pancreatic-specific deletion of Poh1 attenuates ADM and impairs pancreatic carcinogenesis in murine models.\",\n      \"method\": \"Pancreatic-specific conditional Poh1 knockout mouse, co-immunoprecipitation, in vivo ubiquitination assay, ADM murine models\",\n      \"journal\": \"Cancer Letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional KO mouse with in vivo carcinogenesis phenotype, Co-IP, ubiquitination assay\",\n      \"pmids\": [\"37844756\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"POH1 directly interacts with Smad3, removes poly-ubiquitin modifications from Smad3, stabilizes it, and thereby facilitates TGF-β1-mediated lung cancer cell invasion and metastasis.\",\n      \"method\": \"Co-immunoprecipitation, colocalization analysis, in vitro deubiquitination, half-life assay, functional migration/invasion assays, xenograft liver metastasis model\",\n      \"journal\": \"Cancer Letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, in vitro deubiquitination, functional rescue, single lab\",\n      \"pmids\": [\"38061486\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"PSMD14 stabilizes ERα by removing K48-linked polyubiquitin chains, thereby maintaining ERα protein levels and ERα transcriptome in breast cancer. ERα reciprocally binds the PSMD14 promoter to promote its transcription, forming a positive feedback loop. In endocrine-resistant models, PSMD14 inhibition destabilizes the resistant ERα Y537S mutant and restores tamoxifen sensitivity.\",\n      \"method\": \"siRNA DUB library screen, co-immunoprecipitation, K48-specific ubiquitination assay, ChIP assay, endocrine-resistance model, in vivo xenograft\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — DUB screen, linkage-specific ubiquitination, ChIP, resistant mutant model, in vivo validation\",\n      \"pmids\": [\"38017133\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"PSMD14 deubiquitinates β-catenin by decreasing its K48-linked ubiquitination, preventing β-catenin proteasomal degradation and stabilizing it, thereby promoting GBM cell proliferation and invasion. PSMD14 directly interacts with β-catenin.\",\n      \"method\": \"Co-immunoprecipitation, K48-specific ubiquitination assay, siRNA knockdown, rescue with β-catenin overexpression, xenograft mouse model\",\n      \"journal\": \"BioFactors\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, linkage-specific ubiquitination, functional rescue, single lab\",\n      \"pmids\": [\"38696072\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RPN11/PSMD14 deubiquitinates and stabilizes METTL3, an m6A RNA methyltransferase. Stabilized METTL3 enhances m6A modification and expression of ACSS3, which generates propionyl-CoA to upregulate lipid metabolism genes via histone propionylation. Hepatocyte-specific RPN11 knockout mice are protected from diet-induced liver steatosis, insulin resistance, and steatohepatitis.\",\n      \"method\": \"Hepatocyte-specific conditional RPN11 knockout mice, co-immunoprecipitation, ubiquitination assays, m6A sequencing, metabolomics, in vivo diet-induced NAFLD models, pharmacological inhibition (capzimin)\",\n      \"journal\": \"Cell Metabolism\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional KO mouse, Co-IP, ubiquitination assay, metabolomics, m6A sequencing, pharmacological validation\",\n      \"pmids\": [\"39146936\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Rpn11 functions as an allosteric ubiquitin sensor at the 26S proteasome. After substrate recruitment, ubiquitin binding to Rpn11 interferes with conformation-specific interactions of the ubiquitin receptor Rpn10, thereby stabilizing the engagement-competent proteasome state and expediting substrate insertion into the ATPase motor. This allosteric mechanism allows poly-ubiquitin chains or multiple mono-ubiquitins to promote up to 4-fold faster proteasomal turnover.\",\n      \"method\": \"Biochemical assays, active-site and interface mutagenesis, single-molecule FRET (smFRET) measurements of proteasome conformational states, reconstituted degradation assays\",\n      \"journal\": \"Cell Reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — smFRET, mutagenesis, reconstituted biochemistry, multiple orthogonal methods establishing allosteric mechanism\",\n      \"pmids\": [\"40411784\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PSMD14 prevents SLC7A11 ubiquitination and proteasomal degradation by binding to it; glucocorticoids impede this PSMD14-SLC7A11 interaction, causing SLC7A11 degradation, cystine insufficiency, and osteocyte ferroptosis in GIOP. Bone-targeting AAV-mediated PSMD14 overexpression stabilizes SLC7A11 and attenuates both osteocyte ferroptosis and bone loss.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays, AAV-mediated overexpression in mouse model, GPX4 conditional KO, pharmacological ferroptosis inhibitors, high-throughput virtual screening\",\n      \"journal\": \"Advanced Science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assay, in vivo mouse model, single lab\",\n      \"pmids\": [\"40444470\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PSMD14 directly binds and deubiquitinates LDHA to stabilize it, increasing intracellular lactate, which elevates histone lactylation (H3K18la) to transcriptionally activate ACLY and promote lipid biosynthesis and pancreatic cancer progression.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays, histone lactylation measurements, ACLY expression analysis, patient-derived xenograft (PDX) models\",\n      \"journal\": \"Advanced Science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assay, epigenetic readout, PDX validation, single lab\",\n      \"pmids\": [\"41051446\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PSMD14 directly deubiquitinates BCKDK, antagonizing TRIM21-mediated proteasomal degradation, thereby stabilizing BCKDK and promoting SLC7A5/SLC7A8-mediated branched-chain amino acid (BCAA) uptake by GBM cells, leading to NK cell BCAA depletion and immune suppression.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays, PSMD14 knockdown/pharmacological inhibition (OPA), NK cytotoxicity assays, preclinical GBM models\",\n      \"journal\": \"Cell Death & Differentiation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assay, functional immune cell assays, in vivo validation, single lab\",\n      \"pmids\": [\"41876842\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PSMD14 stabilizes IMPDH2, the rate-limiting enzyme of purine nucleotide biosynthesis, by selectively removing K48-linked polyubiquitin chains. PSMD14 inhibition decreases IMPDH2 stability, impairs nucleotide metabolism, causes mitochondrial dysfunction, and increases DNA damage signaling in GBM. Exogenous guanosine reverses these effects.\",\n      \"method\": \"Immunoprecipitation-coupled mass spectrometry (IP-MS), K48-specific ubiquitination assays, metabolic assays, mitochondrial function assessments, guanosine rescue experiments, orthotopic GBM mouse models\",\n      \"journal\": \"Theranostics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — IP-MS, linkage-specific ubiquitination, metabolic rescue experiment, in vivo validation, single lab\",\n      \"pmids\": [\"41608571\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PSMD14 stabilizes PD-L1 by interacting with its intracellular domain and removing K48-linked polyubiquitin chains, thereby inhibiting proteasomal degradation of PD-L1. PSMD14 inhibition promotes PD-L1 degradation, enhances CD8+ T cell activation, reduces Tregs and MDSCs, and improves immunotherapy response in breast cancer syngeneic models.\",\n      \"method\": \"Co-immunoprecipitation, K48-specific ubiquitination assay, cycloheximide chase assay, flow cytometry, T cell cytotoxicity assay, syngeneic mouse models\",\n      \"journal\": \"Journal of Experimental & Clinical Cancer Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, linkage-specific ubiquitination, CHX chase, in vivo syngeneic model, single lab\",\n      \"pmids\": [\"41981629\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PSMD14 deubiquitinates SF3B4 (K63-linked ubiquitin removal), stabilizing SF3B4, which then forms a complex with HNRNPC to promote exon inclusion in FADS1 mRNA via m6A-HNRNPC recognition, upregulating FADS1 and activating Akt/mTOR signaling in TNBC.\",\n      \"method\": \"Co-IP, K63-specific deubiquitination assays, RNA splicing analysis, m6A-RIP, TNBC organoid (PDO) and PDX models\",\n      \"journal\": \"Science Advances\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, linkage-specific ubiquitination, RNA mechanistic assays, PDO/PDX validation, single lab\",\n      \"pmids\": [\"40344056\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"TXNL1's C-terminal tail covers the catalytic groove of the Rpn11 deubiquitinase and coordinates the active-site Zn2+ specifically in substrate-degrading proteasome conformational states (but not in resting state), as revealed by time-resolved cryo-EM. This is consistent with TXNL1 binding to the actively processing proteasome and potentially modulating Rpn11 activity.\",\n      \"method\": \"Time-resolved cryo-EM at saturating and sub-stoichiometric TXNL1 concentrations, biophysical binding experiments\",\n      \"journal\": \"bioRxiv (preprint)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — high-resolution cryo-EM structure, preprint not yet peer-reviewed, single lab\",\n      \"pmids\": [\"bio_10.1101_2024.11.08.622731\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PSMD14/Rpn11 functions non-enzymatically as a receptor for the midnolin ubiquitin-like (Ubl) domain in the midnolin-proteasome pathway (ubiquitin-independent degradation). Cryo-EM structures of the MIDN-bound proteasome show the midnolin Ubl domain binding to Rpn11 (not cleaved by it), positioning the substrate-binding Catch domain above the proteasomal channel. This is distinct from Rpn11's canonical deubiquitinase function.\",\n      \"method\": \"Cryo-EM structures of MIDN-bound human proteasome in two conformational states\",\n      \"journal\": \"bioRxiv (preprint)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — cryo-EM structure, preprint, corroborated by independent parallel preprint (bio_10.1101_2025.03.04.641557)\",\n      \"pmids\": [\"bio_10.1101_2025.02.22.639686\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In the ubiquitin-independent ODC degradation pathway, Rpn11's JAMM motif guides the ODC C-tail into the AAA+ ATPase ring, acting as a translocation gateway and repurposing Rpn11 for a novel ubiquitin-independent function distinct from its canonical deubiquitinase role.\",\n      \"method\": \"Cryo-EM (eleven structures of human 26S proteasome-ODC complexes capturing full degradation process), mutagenesis\",\n      \"journal\": \"bioRxiv (preprint)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — multi-state cryo-EM capturing complete substrate processing, structural basis for non-canonical function, preprint\",\n      \"pmids\": [\"bio_10.1101_2025.11.15.688597\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PSMD14 deubiquitinates CARM1 (coactivator-associated arginine methyltransferase 1), stabilizing it. Stabilized CARM1 activates transcription of FERMT1 through H3R17 dimethylation, promoting HCC proliferation and metastasis.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays, ChIP assay, in vitro and in vivo cell proliferation/metastasis models\",\n      \"journal\": \"Cell Death & Disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assay, ChIP, in vivo validation, single lab\",\n      \"pmids\": [\"40016178\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PSMD14 deubiquitinates RBM15B, preventing its ubiquitin-mediated degradation. Stabilized RBM15B promotes m6A modification of SPON2 mRNA and its stability, facilitating pancreatic cancer progression. PSMD14 itself is transcriptionally activated by MEF2A.\",\n      \"method\": \"ChIP assay (MEF2A-PSMD14 promoter), dual luciferase, co-immunoprecipitation, ubiquitination assay, RIP assay (RBM15B-SPON2 mRNA), MeRIP (m6A on SPON2), subcutaneous and lung metastasis mouse models\",\n      \"journal\": \"Kaohsiung Journal of Medical Sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple molecular assays, in vivo validation, single lab\",\n      \"pmids\": [\"40066751\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PSMD14 deubiquitinates PFKFB2 at K355, facilitating SCYL2-mediated phosphorylation of PFKFB2, which increases fructose-2,6-bisphosphate generation, activates PFK1, and promotes glycolysis and H3K27 lactylation in gastric adenocarcinoma. H3K27 lactylation in turn activates PSMD14 and SOX9 expression, forming a positive feedback loop.\",\n      \"method\": \"Co-immunoprecipitation, site-specific ubiquitination assay (K355), phosphorylation assays, glycolysis metabolic assays, H3K27 lactylation ChIP, FDA drug screening for PSMD14 inhibitors\",\n      \"journal\": \"Cell Death & Differentiation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — site-specific ubiquitination, metabolic assays, epigenetic readout, single lab\",\n      \"pmids\": [\"41193870\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PSMD14 stabilizes FOXM1 by reducing K63-linked ubiquitination on FOXM1, promoting breast cancer progression. PSMD14 also activates the PI3K/AKT/mTOR pathway.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, K63-specific in vitro and in vivo deubiquitination assays, xenograft mouse model\",\n      \"journal\": \"International Journal of Biological Macromolecules\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, linkage-specific ubiquitination, in vivo validation, single lab\",\n      \"pmids\": [\"40902741\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PSMD14 deubiquitinates and stabilizes CDC42 in endothelial cells, promoting filopodia formation and cell migration. Silencing PSMD14 impairs filopodia formation, migration, and CDC42 stability.\",\n      \"method\": \"Co-immunoprecipitation (PSMD14-CDC42 interaction), CDC42 stability/ubiquitination assays, siRNA knockdown, filopodia imaging, migration assays, mouse hindlimb ischemia model\",\n      \"journal\": \"Angiogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, functional loss-of-function, in vivo model, single lab\",\n      \"pmids\": [\"40833626\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Histone lactylation (H3K18la) upregulates PSMD14 transcription after traumatic brain injury. PSMD14 then deubiquitinates PKM2, maintaining PKM protein stability and enabling PSMD14-mediated mitophagy via PINK1 phosphorylation at Thr257, thereby suppressing neuron PANoptosis.\",\n      \"method\": \"LC-MS proteomic analysis, co-immunoprecipitation, ubiquitination assays, PINK1 phosphorylation assays, mitophagy flux analysis, controlled cortical impact (CCI) mouse model\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assay, in vivo CCI model, mechanistic pathway validated, single lab\",\n      \"pmids\": [\"40000916\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"DUSP4-mediated dephosphorylation of PSMD14 enhances PSMD14's interaction with MICALL2 and its deubiquitination activity toward MICALL2, stabilizing MICALL2 and promoting ccRCC malignant progression.\",\n      \"method\": \"Co-immunoprecipitation, MICALL2 ubiquitination assay, PSMD14 phosphorylation analysis, siRNA knockdown, in vivo ccRCC mouse model\",\n      \"journal\": \"Journal of Translational Medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination and phosphorylation assays, in vivo model, single lab\",\n      \"pmids\": [\"42174657\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"PSMD14/POH1/Rpn11 is an essential zinc-dependent JAMM-family metalloprotease deubiquitinase that resides in the lid subcomplex of the 19S proteasome regulatory particle, where it couples substrate deubiquitination to ATP-dependent degradation by cleaving poly-ubiquitin chains en bloc at the base upon mechanical substrate translocation; its catalytic activity requires an intact JAMM motif and is allosterically regulated by ubiquitin binding (promoting proteasome engagement) and by ATPase-driven conformational switching; beyond the proteasome, PSMD14 deubiquitinates a wide array of non-histone substrates (e.g., E2F1, SNAIL, GRB2, TGF-β receptors, pro-IL-1β, ErbB2, ERα, β-catenin, METTL3, PKM2, ALK2, MYC, LDHA, Smad3) to regulate their stability independently of degradation, controls K63-linked ubiquitin chains at DNA double-strand break sites to regulate DNA repair pathway choice, acts as an epigenetic H2AK119 deubiquitinase on chromatin with NSD2, and regulates Golgi-to-ER retrograde transport and macroautophagy.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"PSMD14 (POH1/Rpn11) is the essential zinc-dependent JAMM-family metalloprotease deubiquitinase of the 19S proteasome lid that couples substrate deubiquitination to ATP-dependent proteolysis, removing poly-ubiquitin chains en bloc as substrates are translocated into the degradation machinery [#0, #2]. Its catalytic activity resides in the MPN+/JAMM motif, whose conserved active-site residues are required for viability in yeast and human cells and for proteasomal degradation; active-site mutation abolishes deubiquitination and is lethal [#0, #1, #5]. Structural and biochemical work established that Rpn11 is a promiscuous, low-affinity deubiquitinase lacking ubiquitin-linkage specificity at the level of the isolated MPN domain, and that full activation requires incorporation into the 26S proteasome and ATP hydrolysis—premature activity being prevented by an Insert-1 barrier loop and an unstable catalytic loop [#11, #12]. Substrate-driven ubiquitin binding switches the Ins-1 loop to an active conformation in a step that is rate-limiting and accelerated by mechanical translocation, placing Rpn11 in direct kinetic competition with ATPase-driven ubiquitin unfolding [#15]; ubiquitin binding also acts allosterically through Rpn10 to stabilize the engagement-competent proteasome state and speed turnover [#34]. Within and beyond the proteasome, PSMD14 preferentially processes K63-linked chains [#6, #13], a specificity it exploits at sites of DNA double-strand breaks, where it removes RNF8/RNF168-generated K63 chains to limit 53BP1 accumulation, counter the RAP80 barrier to end resection, and promote homologous recombination [#9, #10]. Loss of PSMD14 causes polyubiquitin accumulation, G0/G1 arrest and senescence, distinguishing 19S from 20S function [#5, #8]. Independently of bulk degradation, PSMD14 deubiquitinates and stabilizes numerous substrates—E2F1, SNAIL, TGF-\\u03b2 receptors and Smad3, ALK2, ER\\u03b1, \\u03b2-catenin, MYC, GRB2 and others—removing K48- or K63-linked chains to control their stability and downstream signaling, frequently in cancer contexts and often within transcriptional positive-feedback loops [#14, #18, #23, #22, #31, #29, #30]. PSMD14 further acts on chromatin as a non-proteasomal H2AK119 deubiquitinase in complex with NSD2 to license H3K36 methylation and transcription [#28], regulates K63-dependent Golgi-to-ER retrograde transport and macroautophagy [#24], and is targeted by zinc-chelating inhibitors including capzimin and thiolutin that stabilize proteasome substrates and induce the unfolded protein response [#16, #17].\",\n  \"teleology\": [\n    {\n      \"year\": 2002,\n      \"claim\": \"Establishing whether the proteasome lid contains an intrinsic deubiquitinase resolved how ubiquitin is removed during degradation: Rpn11 was identified as a JAMM metalloprotease whose activity is mechanistically coupled to substrate proteolysis.\",\n      \"evidence\": \"Active-site mutagenesis (rpn11AXA) with yeast lethality and in vitro reconstituted degradation of ubiquitinated Sic1; MPN+ motif mutagenesis with proteolysis defects\",\n      \"pmids\": [\"12183636\", \"12370088\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve the structural basis of substrate engagement\", \"Linkage specificity not yet defined\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Defining how proteasomal deubiquitination is partitioned showed Rpn11 and Ubp6 play complementary, non-redundant roles and confirmed Rpn11 as a metal-dependent enzyme requiring an intact lid for coupling to degradation.\",\n      \"evidence\": \"Proteasome purification from yeast mutants, in vitro DUB assays, metal chelator sensitivity, synthetic lethality genetics\",\n      \"pmids\": [\"14581483\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism coupling deubiquitination to translocation not defined\", \"Metal identity inferred, not structurally resolved\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Dissecting Rpn11 into separable domains revealed a catalytic-independent C-terminal function in mitochondrial morphology, indicating roles beyond proteasomal DUB activity.\",\n      \"evidence\": \"Yeast RPN8-RPN11 chimera analysis, intragenic complementation, mitochondrial microscopy, cell cycle analysis\",\n      \"pmids\": [\"15018611\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism of the mitochondrial function unresolved\", \"Not extended to mammalian cells in this work\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Testing requirement in human cells confirmed the JAMM motif is essential for viability and 26S function, validating PSMD14 catalysis as the operative human activity.\",\n      \"evidence\": \"siRNA knockdown with wild-type vs JAMM-mutant rescue, viability and polyubiquitin accumulation assays in HeLa cells\",\n      \"pmids\": [\"17237285\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not address non-proteasomal functions\", \"Substrate scope not examined\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defining linkage specificity and downstream phenotype showed proteasome-associated PSMD14 is a K63-specific metalloDUB and that 19S loss produces cell cycle arrest distinct from 20S inhibition.\",\n      \"evidence\": \"Seven-step chromatographic co-fractionation with linkage-specific assays and inhibitor profiling; siRNA with flow cytometry and comparison to PSMB5\",\n      \"pmids\": [\"19214193\", \"19732767\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"K63 specificity at the isolated enzyme level not yet reconciled with later structural promiscuity\", \"Direct substrates driving arrest not identified\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Early substrate studies hinted that PSMD14 deubiquitinates specific proteins (c-Jun, ErbB2) in a manner not strictly coupled to degradation, foreshadowing degradation-independent stabilization roles.\",\n      \"evidence\": \"Ectopic overexpression and active-site mutagenesis for c-Jun; siRNA DUB screen and Western/flow cytometry for ErbB2\",\n      \"pmids\": [\"16569633\", \"19436748\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab Co-IP/ubiquitination without reciprocal validation\", \"Selectivity claims rest on limited substrate panels\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Positioning PSMD14 in DNA repair established that it processes K63 chains at break sites to limit 53BP1, counter the RAP80 resection barrier, and bias repair toward homologous recombination.\",\n      \"evidence\": \"siRNA knockdown and co-depletion, immunofluorescence of DSB foci, K63-Ub chromatin analysis, survival assays\",\n      \"pmids\": [\"22909820\", \"24013561\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether this is proteasome-dependent or free-DUB activity not fully resolved\", \"Direct chromatin substrates not enumerated\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Crystallographic and biochemical analyses resolved the structural logic of Rpn11: it is an intrinsically promiscuous, low-affinity DUB whose activation is gated by proteasome incorporation and ATP hydrolysis.\",\n      \"evidence\": \"Rpn11-Rpn8 heterodimer crystal structures, nanobody-assisted crystallization, cryo-EM docking, mutagenesis, defined-chain in vitro DUB kinetics\",\n      \"pmids\": [\"24463465\", \"24516147\", \"25389291\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The ubiquitin-bound active conformation not yet captured at this stage\", \"Reconciliation with cellular K63 preference incomplete\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"An in vivo substrate paradigm showed PSMD14 stabilizes E2F1 to drive oncogenic transcription, exemplifying degradation-independent substrate control in mammals.\",\n      \"evidence\": \"Reciprocal Co-IP, in vivo ubiquitination, conditional Poh1 knockout mouse liver, xenografts\",\n      \"pmids\": [\"26510456\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether stabilization is proteasome-associated or free-enzyme not dissected\", \"Linkage type on E2F1 not specified\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Capturing the ubiquitin-bound state and developing selective inhibitors defined the mechanochemical activation switch and validated PSMD14 as a druggable target competing with ATPase-driven ubiquitin unfolding.\",\n      \"evidence\": \"Ubiquitin-bound Rpn11 crystal structure with single-molecule and ensemble DUB assays; capzimin and thiolutin zinc-chelation inhibition with proteomics and cell viability\",\n      \"pmids\": [\"28844860\", \"28244987\", \"28459440\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Inhibitor selectivity over related JAMM proteases is limited\", \"In vivo therapeutic window not established in these studies\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Expanding the substrate repertoire established broad degradation-independent stabilization activity across SNAIL, TGF-\\u03b2 receptors, ALK2 and GRB2, frequently via removal of specific K48 or K63 chains in cancer.\",\n      \"evidence\": \"Mass spectrometry interactomics, Co-IP, linkage-specific ubiquitination assays, conditional knockout mice, xenograft and metastasis models\",\n      \"pmids\": [\"29331416\", \"30315153\", \"30745168\", \"31634528\", \"31685442\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Most substrate studies are single-lab without reciprocal cross-validation\", \"Whether substrate DUB activity requires proteasome incorporation often untested\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"A trafficking and autophagy role showed PSMD14 acts on cellular K63 chains to enable Golgi-to-ER retrograde transport, distinct from 20S proteasome function.\",\n      \"evidence\": \"High-content siRNA ubiquitinome screen, capzimin treatment, trafficking-marker microscopy, autophagy flux, 19S vs 20S comparison\",\n      \"pmids\": [\"32210007\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct K63-ubiquitinated trafficking substrates not identified\", \"Mechanism of recruitment to Golgi unresolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Demonstrating a chromatin-resident, proteasome-independent activity revealed PSMD14 as an H2AK119 deubiquitinase in an NSD2 complex that licenses transcription, establishing a non-proteasomal epigenetic role.\",\n      \"evidence\": \"ChIP-seq, chromatin Co-IP, histone H2AK119 deubiquitination assays, integrative epigenomics, inhibitor studies\",\n      \"pmids\": [\"37935198\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How PSMD14 is partitioned between proteasome and chromatin pools unclear\", \"Generality across cell types beyond myeloma not established\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Mechanistic refinement defined PSMD14 as an allosteric ubiquitin sensor that accelerates proteasomal turnover via Rpn10, while a wave of substrate studies (MYC, ER\\u03b1, METTL3, LDHA, \\u03b2-catenin, PD-L1, BCKDK, IMPDH2, metabolic and immune regulators) extended its degradation-independent stabilization activity into metabolism, immunity, and feedback transcriptional loops.\",\n      \"evidence\": \"smFRET, interface mutagenesis and reconstituted degradation for the allosteric mechanism; Co-IP, linkage-specific ubiquitination, conditional knockouts and PDX/syngeneic models for substrates\",\n      \"pmids\": [\"40411784\", \"37844756\", \"38017133\", \"39146936\", \"41051446\", \"38696072\", \"41981629\", \"41876842\", \"41608571\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Substrate studies are predominantly single-lab and context-specific\", \"Whether individual substrate DUB events occur at the proteasome or by a free enzyme pool largely untested\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Structural studies began to reveal non-canonical, ubiquitin-independent uses of Rpn11—as a midnolin Ubl receptor and an ODC translocation gateway—and a TXNL1-mediated regulatory contact in processing-state proteasomes.\",\n      \"evidence\": \"Time-resolved and multi-state cryo-EM of TXNL1-, MIDN-, and ODC-bound proteasomes (preprints)\",\n      \"pmids\": [\"bio_10.1101_2024.11.08.622731\", \"bio_10.1101_2025.02.22.639686\", \"bio_10.1101_2025.11.15.688597\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprints not yet peer-reviewed\", \"Functional consequences in cells not established\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how PSMD14 is physically and functionally partitioned between its proteasome-bound deubiquitinase role and its numerous proteasome-independent activities on individual substrates and chromatin, and what governs substrate selection at the cellular level.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No systematic distinction between proteasome-associated and free-enzyme substrate processing\", \"Recruitment determinants for specific substrates unknown\", \"Quantitative cellular pools of proteasomal vs non-proteasomal PSMD14 not measured\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 2, 5, 6, 13]},\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [0, 6, 16]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [34]},\n      {\"term_id\": \"GO:0140299\", \"supporting_discovery_ids\": [34]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [28, 9]},\n      {\"term_id\": \"GO:0000228\", \"supporting_discovery_ids\": [9, 28]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [24]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 2, 5]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [9, 10]},\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [28]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [24, 27]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [24]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"complexes\": [\n      \"19S proteasome regulatory particle (lid)\",\n      \"26S proteasome\",\n      \"PSMD14-NSD2 chromatin complex\"\n    ],\n    \"partners\": [\n      \"PSMD7\",\n      \"PSMD10\",\n      \"NSD2\",\n      \"E2F1\",\n      \"SNAIL\",\n      \"TGFBR1\",\n      \"MYC\",\n      \"TXNL1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}