Affinage

USP28

Ubiquitin carboxyl-terminal hydrolase 28 · UniProt Q96RU2

Length
1077 aa
Mass
122.5 kDa
Annotated
2026-06-11
84 papers in source corpus 48 papers cited in narrative 47 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 9/9 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

USP28 is a constitutively active, dimeric ubiquitin-specific protease that controls cell proliferation, oncoprotein abundance, and a p53-dependent mitotic surveillance checkpoint by reversing proteasomal degradation of selected substrates (PMID:24960159, PMID:30926242, PMID:30926243). Its founding role is the protection of oncoproteins from SCF(FBW7)-mediated turnover: USP28 binds the nucleoplasmic FBW7alpha isoform and stabilizes c-MYC, and dissociates from FBW7 upon DNA damage to permit MYC decline (PMID:17558397, PMID:17873522), and in vivo it antagonizes FBW7-directed degradation of c-MYC, c-JUN, and NOTCH1 in intestinal proliferation and lineage differentiation (PMID:24960159). Mechanistically, USP28 recognizes the same phosphodegron motifs targeted by FBW7 (in their unphosphorylated state) and also stabilizes FBW7 itself by reversing its autoubiquitination, so that both loss and excess of USP28 perturb FBW7 substrate balance and promote oncogenic transformation (PMID:25716680, PMID:25437563). In a second, FBW7-independent arm, USP28 forms a 53BP1-dependent module that directly deubiquitinates and stabilizes p53 to arrest growth after centrosome loss or prolonged mitosis, a mitotic surveillance pathway operating in parallel to the spindle assembly checkpoint and active in the developing brain (PMID:27432897, PMID:27432896, PMID:27371829, PMID:33226141). The 53BP1 interaction is mediated by the USP28 C-terminal domain in a PLK1- and dimerization-dependent manner; cancer-associated missense mutations in this domain disrupt 53BP1 binding, impair nuclear localization, and destabilize USP28 (PMID:41365927). USP28 activity is set by its oligomeric state—its dimerizing insertion spatially separates two independently active catalytic domains, distinguishing it from the autoinhibited tetramer of its paralog USP25—and dimerization both limits MYC-driven transcriptional elongation and is selectively bound and stabilized by 53BP1 (PMID:30926242, PMID:30926243, PMID:38227944). This activity is further tuned by post-translational regulation: N-terminal SUMOylation is inhibitory and SENP1-mediated deSUMOylation is activating under hypoxia (PMID:25359778, PMID:30622440), ATR phosphorylation increases activity during DNA damage (PMID:34611298), nuclear caspase-8 cleaves and inactivates USP28 to abolish p53 stabilization (PMID:31982308), and DYRK2 phosphorylation and DTX3L ubiquitination drive its degradation in reciprocal regulatory loops (PMID:40858801, PMID:40703443). USP28 is required for induction and maintenance of squamous cell carcinoma through stabilization of ΔNp63, and genetic or small-molecule inhibition of USP28 lowers c-MYC, c-JUN, and ΔNp63 and causes regression of lung tumors, establishing it as a tractable cancer target (PMID:32128997, PMID:34636321). Beyond these themes, a broad set of substrates extends USP28's deubiquitinase activity across additional oncogenic, metabolic, and stress pathways (PMID:24075993, PMID:34584067, PMID:37202505, PMID:37994595, PMID:39431010).

Mechanistic history

Synthesis pass · year-by-year structured walk · 15 steps
  1. 2001 Medium

    Establishing USP28 as a distinct deubiquitinase gene defined the molecular class and the existence of tissue-specific isoforms before any function was known.

    Evidence homology search and cDNA cloning at 11q23 with isoform characterization

    PMID:11597335

    Open questions at the time
    • No catalytic activity or substrate demonstrated
    • Functional significance of tissue-specific isoforms not tested
  2. 2006 High

    Discovery of USP28 within 53BP1 complexes answered what links it to the DNA damage response, showing it stabilizes checkpoint factors and enables damage-induced apoptosis.

    Evidence Co-IP of 53BP1 complexes, RNAi knockdown, apoptosis assays in cell lines

    PMID:16901786

    Open questions at the time
    • Direct deubiquitination of named checkpoint substrates not shown in vitro
    • Mechanism of 53BP1 recruitment not yet defined
  3. 2007 High

    Identification of the USP28-FBW7alpha interaction answered how MYC is protected from degradation and tied the protease to oncoprotein stability in a subcellularly compartmentalized manner.

    Evidence shRNA screen, reciprocal Co-IP, subcellular fractionation, knockdown/rescue in tumor cells

    PMID:17558397 PMID:17873522

    Open questions at the time
    • Whether USP28 acts directly on MYC ubiquitin chains versus via FBW7 not fully resolved
    • Structural basis of FBW7alpha selectivity unknown
  4. 2014 High

    In vivo deletion and double-knockout genetics defined USP28 as an FBW7 antagonist that recognizes the same phosphodegron motifs and also stabilizes FBW7 itself, explaining its dual oncogenic/tumor-suppressive behavior.

    Evidence Usp28 knockout and Usp28/Fbw7 double-knockout mice, Ras transformation assays, phosphodegron interaction assays

    PMID:24960159 PMID:25437563 PMID:25716680

    Open questions at the time
    • Determinants distinguishing FBW7-dependent from FBW7-independent substrates not defined
    • Quantitative basis of the dose-dependent transformation phenotype unclear
  5. 2014 High

    Biochemistry of USP28 regulation and chain specificity answered how its activity is restrained, showing inhibitory N-terminal SUMOylation and preference for K11/K48/K63 chains.

    Evidence in vitro deubiquitination assays, SUMO modification, N-terminal truncation and linkage-specificity assays

    PMID:25359778

    Open questions at the time
    • Physiological substrates whose chains are SUMO-regulated not mapped
    • SUMO sites and writer/eraser enzymes not identified in this study
  6. 2016 High

    Convergent CRISPR screens placed USP28 in a 53BP1-p53 mitotic surveillance pathway, answering how cells arrest after centrosome loss or prolonged mitosis independently of DNA repair.

    Evidence genome-wide CRISPR centrinone-resistance screens in multiple labs, in vitro p53 deubiquitination, genetic epistasis

    PMID:27371829 PMID:27432896 PMID:27432897

    Open questions at the time
    • How mitotic delay is sensed and transmitted to USP28 not defined
    • Relationship between this pathway and FBW7-substrate functions unresolved
  7. 2019 High

    Crystal structures answered the basis of USP28's constitutive activity, showing it forms an active dimer rather than the autoinhibited tetramer of USP25, and that oligomeric state tunes substrate stabilization.

    Evidence crystal structures of catalytic domains, in vitro activity and oligomerization assays, cell-based substrate stabilization, companion papers

    PMID:30926242 PMID:30926243

    Open questions at the time
    • How cellular signals shift oligomeric state was not addressed here
    • Structure of full-length protein with regulatory domains not determined
  8. 2019 Medium

    Demonstrating SENP1-mediated deSUMOylation under hypoxia answered how USP28 activity is switched on to stabilize HIF-1alpha, defining a positive feedback loop.

    Evidence Co-IP, GST pulldown, in vitro deubiquitination, SUMO-deficient mutants, luciferase reporters

    PMID:30622440

    Open questions at the time
    • In vivo relevance of the SENP1-USP28-HIF1a loop not tested
    • Single lab
  9. 2020 High

    Identification of ΔNp63 as a substrate and caspase-8 as an inactivating protease answered how USP28 maintains squamous carcinoma identity and how its tumor-suppressive p53 arm can be silenced in cancer.

    Evidence in vivo CRISPR SCC mouse models, pharmacological/genetic inhibition, nuclear caspase-8 cleavage and p53 stability assays

    PMID:31982308 PMID:32128997

    Open questions at the time
    • Trigger and regulation of nuclear caspase-8 activity not defined
    • Whether caspase-8 cleavage affects FBW7-substrate arm not tested
  10. 2021 High

    DNA-damage signaling to USP28 was clarified by showing ATR phosphorylation activates it and links damage repair to ΔNp63 stability in SCC, and pharmacological inhibition validated USP28 as a druggable target.

    Evidence ATR inhibitor and phosphorylation assays, recruitment to damage sites; small-molecule USP28 inhibitor causing tumor regression that phenocopies genetic deletion

    PMID:34611298 PMID:34636321

    Open questions at the time
    • ATR phosphosites on USP28 not all mapped
    • Selectivity of the inhibitor versus USP25 not fully resolved in this context
  11. 2023 Medium

    Dimerization mutant and ChIP analyses answered how USP28 oligomeric state couples to transcription, showing monomeric USP28 stabilizes MYC and drives PAF1c recruitment and replication stress, while 53BP1 selectively binds and stabilizes dimers.

    Evidence dimerization mutants, ChIP, DNA synthesis and damage assays, Co-IP with 53BP1

    PMID:38227944

    Open questions at the time
    • What physiologically drives monomer-dimer switching in cells
    • Single lab
  12. 2024 High

    Mapping the C-terminal, PLK1-dependent 53BP1-binding domain and characterizing cancer mutations answered how the mitotic surveillance complex assembles and how mutations cripple p53 stabilization.

    Evidence cancer mutation analysis, domain interaction mapping, nuclear localization and p53 deubiquitination assays, Co-IP, cell cycle assays

    PMID:41365927

    Open questions at the time
    • How PLK1 phosphorylation is timed within mitosis not detailed
    • Effect of mutations on FBW7-substrate functions not assessed
  13. 2024 High

    Inhibitor co-crystal structures answered the structural basis of pharmacological USP28 inhibition, identifying a common binding pocket and a key glutamate critical for activity.

    Evidence crystal structures with AZ1, Vismodegib, FT206; E366 mutagenesis and inhibitor binding assays

    PMID:38816515

    Open questions at the time
    • USP25/USP28 selectivity of the pocket not fully resolved
    • Allosteric coupling to dimerization not addressed
  14. 2025 Medium

    Identification of DTX3L and DYRK2 as enzymes that degrade USP28, with reciprocal USP28 stabilization of each, answered how USP28 abundance is fine-tuned and feeds back on DSB repair and p53 signaling.

    Evidence Co-IP, ubiquitination/phosphorylation assays, DSB repair pathway assays, kinase activity and domain mapping

    PMID:40703443 PMID:40858801

    Open questions at the time
    • Hierarchy among caspase-8, DYRK2, and DTX3L control of USP28 not defined
    • Single labs
  15. 2025 Medium

    Chemical-biology disruption of the 53BP1-USP28 interface answered whether USP28 catalytic activity sustains the complex, showing catalytic activity maintains 53BP1 binding and that USP28 negatively regulates p53 transactivation in wild-type settings.

    Evidence crystal structure with CAS-010 inhibitor, 53BP1 interaction and p53 transactivation assays, catalytic-mutant analysis

    PMID:40902594

    Open questions at the time
    • Reconciliation with prior models where USP28 stabilizes p53 not fully resolved
    • Single lab

Open questions

Synthesis pass · forward-looking unresolved questions
  • It remains unclear what governs the choice between USP28's FBW7-substrate stabilization arm and its 53BP1-p53 surveillance arm, and how its many post-translational regulators are integrated to set substrate selectivity in a given cell state.
  • No unified model linking oligomeric state, PTMs, and substrate choice
  • Limited cross-substrate prioritization data
  • Many substrate claims rest on single-lab Co-IP studies

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 4 GO:0098772 molecular function regulator activity 3 GO:0016787 hydrolase activity 2
Localization
GO:0005634 nucleus 2 GO:0005654 nucleoplasm 1
Pathway
R-HSA-1640170 Cell Cycle 3 R-HSA-392499 Metabolism of proteins 3 R-HSA-73894 DNA Repair 3 R-HSA-1643685 Disease 2
Partners
CASP8DTX3LDYRK2FBXW7MYCSENP1TP53TP53BP1
Complex memberships
53BP1-USP28 mitotic surveillance complex

Evidence

Reading pass · 47 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2007 USP28 binds to MYC through an interaction with FBW7alpha (nucleoplasmic isoform), stabilizing MYC in the nucleus but not in the nucleolus where FBW7gamma mediates MYC degradation. USP28 is required for MYC stability in human tumor cells. shRNA screen, Co-IP, subcellular fractionation, knockdown/rescue experiments Nature cell biology High 17558397
2006 USP28 is required to stabilize Chk2 and 53BP1 in response to DNA damage (double-strand breaks). USP28 was discovered through analysis of 53BP1 complexes and is required for DNA-damage-induced apoptosis through the Chk2-p53-PUMA pathway. USP28 also stabilizes Mdc1, Claspin, and TopBP1 in the DNA-damage checkpoint pathway. Co-IP (53BP1 complex analysis), cell line functional assays, RNAi knockdown, apoptosis assays Cell High 16901786
2007 Usp28 dissociates from Fbw7alpha in response to UV irradiation, providing a mechanism by which Fbw7-mediated degradation of Myc is enhanced upon DNA damage. Myc decline after DNA damage is mediated by proteasomal degradation via the Fbw7 ubiquitin ligase. Co-IP, UV irradiation, protein stability assays, proteasome inhibitor experiments Cell cycle (Georgetown, Tex.) Medium 17873522
2011 USP28 antagonizes GSK-3β/Fbw7-dependent HIF-1α degradation. Fbw7 and USP28 reciprocally regulate cell migration and angiogenesis in an HIF-1α-dependent manner. GSK-3β and Fbw7-deficient cell experiments, overexpression/knockdown, migration and angiogenesis assays Blood Medium 22144179
2013 USP28 is a deubiquitinase of LSD1. USP28 directly interacts with LSD1 and stabilizes it via deubiquitination, counteracting its proteasome-mediated degradation. Knockdown of USP28 results in LSD1 destabilization and suppression of cancer stem cell-like characteristics rescued by ectopic LSD1 expression. siRNA screen against all human deubiquitinases, Co-IP, deubiquitination assay, rescue experiments, in vivo xenograft Cell reports High 24075993
2014 USP28 antagonizes c-MYC, c-JUN, and NOTCH1 ubiquitin-dependent degradation in the intestine. Usp28 deletion in mice reduced intestinal proliferation and impaired secretory lineage cell differentiation. USP28 and c-MYC form a positive feedback loop where USP28 is a c-MYC target gene. Murine genetic models (Usp28 knockout), intestinal tumorigenesis models, protein stability assays, mouse lifespan analysis The Journal of clinical investigation High 24960159
2014 USP28 preferentially antagonizes autocatalytic (self-)ubiquitination of Fbw7, stabilizing Fbw7. Monoallelic Usp28 deletion maintains stable Fbw7 and drives Fbw7 substrate degradation, while complete knockout triggers Fbw7 degradation and accumulation of Fbw7 substrates. Both complete loss and overexpression of Usp28 promote Ras-driven oncogenic transformation. Usp28 knockout mouse models (monoallelic and biallelic), embryonic fibroblast assays, Ras transformation assay, protein stability measurements Cell reports High 25437563
2014 USP28 is recruited to double-strand breaks in a manner that requires the tandem BRCT domains of 53BP1. However, USP28 depletion caused only minor DDR defects, and mice lacking USP28 showed normal longevity, immunological development, and radiation responses, indicating USP28 is not a critical factor in double-strand break metabolism. Recruitment assay to DSBs, BRCT domain mutant analysis, USP28-depleted cells, Usp28 knockout mice with radiation challenge Molecular and cellular biology High 24687851
2014 SUMO modification of the N-terminal domain of USP28 negatively regulates its deubiquitinating activity. Truncation of the N-terminal domain does not impair deubiquitinating activity on diubiquitin or polyubiquitin chain substrates. USP28 shows chain preference for Lys11-, Lys48-, and Lys63-linked diubiquitin linkages. In vitro deubiquitination assays, SUMO modification assays, N-terminal truncation mutants, linkage-specificity assays The Journal of biological chemistry High 25359778
2015 Usp28 inactivation in the intestine rescues hyperproliferation and impaired goblet/Paneth cell differentiation of Fbw7-deficient mice. Usp28 deficiency corrects accumulation of SCF(Fbw7) substrate proteins (NICD1, c-Jun, c-Myc) independently of Fbw7 presence. Usp28 interacts with unphosphorylated phosphodegron motifs (the same motif recognized by Fbw7 when phosphorylated), offering a mechanistic explanation for identical substrate selection. Intestinal-specific Usp28/Fbw7 double knockout mice, protein stability assays, phosphodegron interaction assays Cancer research High 25716680
2016 USP28 and 53BP1 form a module acting upstream of p53 to arrest cell growth after centrosome loss or prolonged mitosis. USP28 and 53BP1 stabilize p53 after centrosome loss, independently of their role in the DNA damage response. This USP28-53BP1-p53-p21 signaling axis also arrests growth after prolonged prometaphase. Genome-wide CRISPR/Cas9 knockout screen (centrinone resistance), genetic deletion of TP53BP1 and USP28, p53 protein level assays, cell cycle analysis The Journal of cell biology High 27371829 27432896 27432897
2016 53BP1 mediates p53 activation independently of its DNA repair activity but requiring USP28. USP28 can directly deubiquitinate p53 in vitro and ectopically stabilize p53 in vivo. 53BP1 and USP28 function in parallel to the spindle assembly checkpoint to select against disturbed or delayed mitosis. CRISPR loss-of-function screen, in vitro deubiquitination assay of p53, in vivo p53 stabilization assay, genetic epistasis eLife High 27371829
2018 USP28 functions through a feedback loop to destabilize RAF family members; loss of USP28 stabilizes BRAF, enhances downstream MAPK activation, and promotes resistance to RAF inhibitor therapy. USP28 is deleted in a proportion of melanoma patients. Cell culture and in vivo models, BRAF stability assays, USP28 knockdown/knockout, MAPK pathway activation measurement The Journal of experimental medicine Medium 29880484
2018 LIN28A protein undergoes ubiquitination and USP28 interacts with and stabilizes LIN28A by reversing its proteasomal degradation via deubiquitinating activity. Co-IP, deubiquitination assay, protein stability (cycloheximide chase), knockdown/overexpression Biochimica et biophysica acta. Molecular basis of disease Medium 30543854
2019 USP28 forms a constitutively active dimer through a dimerizing insertion that spatially separates independently active catalytic domains. Unlike USP25, USP28 does not form an autoinhibited tetramer. Oligomeric states of USP28 confirmed in cells and modulating oligomerization affects substrate stabilization in accordance with in vitro activity. Crystal structure (structural determination), biochemical activity assays, cell-based substrate stabilization assays, in vitro oligomerization experiments Molecular cell High 30926242 30926243
2019 USP28 is a constitutively active dimer, while USP25 forms an autoinhibited tetramer; structures of the catalytic domains of both proteins determined. USP28 dimerization is intrinsic to its constitutive activity. Cancer-associated USP25 mutations lead to activation in vitro and in vivo. Crystal structure of catalytic domains, biochemical activity assays, in vitro and in vivo oligomerization studies Molecular cell High 30926243
2019 SENP1-mediated deSUMOylation of USP28 activates USP28 under hypoxia. SUMOylation of USP28 under normoxia limits its deubiquitinating activity toward HIF-1α. Hypoxia activates USP28 through SENP1-mediated deSUMOylation, creating a positive feedback loop to further accumulate HIF-1α. Co-IP, GST pulldown, in vitro deubiquitination assay, in vivo deSUMOylation assay, luciferase reporter assay, SUMOylation-deficient mutant Cancer cell international Medium 30622440
2019 USP28 knockdown caused increased ubiquitination of histone H2A at K119, reduced expression of p53, p21 and p16INK4a, and increased cell proliferation, establishing USP28 as a deubiquitinase for histone H2A. RNAi screen of DUBs, USP28 knockdown, H2A ubiquitination assay, cell proliferation assay Experimental cell research Low 30910399
2020 Nuclear caspase-8 cleaves and inactivates USP28, preventing USP28 from deubiquitinating and stabilizing wild-type p53. This cleavage results in de facto p53 protein loss, switching cell fate from apoptosis toward mitosis, allowing tumor cells to override the p53-dependent G2/M checkpoint. Nuclear caspase-8 localization studies, Co-IP, USP28 cleavage assay, p53 stability assays, genetic depletion with cell cycle/apoptosis readouts Molecular cell High 31982308
2020 The 53BP1-USP28 mitotic surveillance pathway is active in developing brain. Depletion of centrosome proteins in neural progenitor cells prolongs mitosis and increases TP53-mediated apoptosis. Inactivation of the mitotic surveillance pathway (53BP1 or USP28 deletion) restored NPC proliferation and brain size without correcting upstream centrosome defects or extended mitosis. Mouse genetic models (53BP1 and USP28 knockout), centrosome protein depletion in NPCs, apoptosis assays, brain size measurements The EMBO journal High 33226141
2020 USP28 stabilizes ΔNp63 and maintains elevated ΔNp63 levels in squamous cell carcinoma by counteracting its proteasome-mediated degradation. Impaired USP28 activity (genetic or pharmacological) abrogates transcriptional identity and suppresses growth of human SCC cells. CRISPR/Cas9 in vivo mouse models show endogenous USP28 is required for both induction and maintenance of lung SCC. CRISPR/Cas9 in vivo mouse models, siRNA, pharmacological inhibition, protein stability assays, gene expression analysis EMBO molecular medicine High 32128997
2021 ATR phosphorylates USP28 in response to cisplatin treatment, increasing its enzymatic activity. USP28 is recruited to sites of DNA damage in cisplatin-treated cells, and this phosphorylation positively regulates DNA damage repair in SCC by stabilizing ΔNp63. ATR inhibitor studies, phosphorylation assays, USP28 recruitment to DNA damage sites, enzymatic activity assays, knockdown/inhibitor experiments Cell death and differentiation Medium 34611298
2021 USP28 deubiquitinates and stabilizes FOXM1, a critical mediator of Wnt/β-catenin signaling. USP28-mediated stabilization of FOXM1 promotes nucleus β-catenin trans-activation and activation of the Wnt/β-catenin pathway in pancreatic cancer. Co-IP, deubiquitination assay, protein stability assay, rescue experiments with FOXM1 re-expression, in vivo xenograft Cell death & disease Medium 34584067
2021 Genetic inactivation of Usp28 induced regression of established murine LSCC lung tumors. A small-molecule USP28 inhibitor decreased c-MYC, c-JUN, and ΔNp63 protein levels and caused regression of autochthonous murine LSCC and human LSCC xenografts, phenocopying genetic deletion. Genetic mouse tumor model, small-molecule inhibitor, tumor regression analysis, protein stability assays eLife High 34636321
2021 USP28 interacts with and deubiquitinates UCK1, antagonizing KLHL2-mediated polyubiquitination of UCK1 at K81. ATM-mediated phosphorylation of USP28 results in its disassociation from KLHL2 and UCK1 destabilization. Mass spectrometry, Co-IP, ubiquitination assays (identifying K81 site), ATM phosphorylation studies, AML cell line and murine AML model Theranostics Medium 31938050
2022 USP28 interacts with PIM1 and PIM2 kinases, and this interaction increases in response to hypoxia, correlating with reduced PIM ubiquitination and increased protein stability. USP28 overexpression increased PIM protein stability; USP28 knockdown significantly increased ubiquitination of PIM1 and PIM2. Co-IP, ubiquitination assay, overexpression/knockdown, in vivo xenograft Cells Medium 35326457
2022 USP28 interacts with NuMA1 and affects centrosome clustering in tetraploid cells. Loss of USP28 reduces checkpoint activation in tetraploid cells, facilitating their proliferation. Mass spectrometry, immunoprecipitation, RNAi screen, cell viability assays Cellular oncology (Dordrecht, Netherlands) Low 34962618
2023 USP28 directly binds to mature SREBP2, leading to its deubiquitination and stabilization. USP28 silencing reduces expression of mevalonate pathway enzymes and lowers metabolic flux. USP28 deletion rendered cancer cells highly sensitive to MVP inhibition by statins, rescued by geranyl-geranyl pyrophosphate. Co-IP, deubiquitination assay, metabolic flux analysis, CRISPR/Cas9 deletion, tissue microarray, mouse tumor model Cell death and differentiation Medium 37202505
2023 USP28 dimerization limits USP28 activity and restricts recruitment of the elongation factor PAF1c by MYC. Monomeric USP28 stabilizes MYC and promotes PAF1c recruitment, leading to ectopic DNA synthesis and replication-associated DNA damage. 53BP1 selectively binds USP28 dimers and stimulates dimerization; genotoxic stress diminishes 53BP1-USP28 interaction and promotes USP28 dimer disassembly. Dimerization mutants, ChIP, DNA replication assays, DNA damage assays, Co-IP with 53BP1 Nucleic acids research Medium 38227944
2023 The HNF-1β transcription factor directly regulates USP28 expression as a target gene. USP28 in turn stabilizes Claspin (by deubiquitination), which promotes Chk1 activation and G2/M cell cycle progression in response to DNA damage in ovarian clear cell carcinoma. siRNA knockdown, immunoprecipitation (Claspin ubiquitination), RNAi-mediated gene silencing, Chk1 phosphorylation assay Oncotarget Low 29707125
2023 USP28 directly interacts with PPARα, deubiquitinating and stabilizing PPARα (at Lys152), which promotes Mfn2 transcription to impede mitochondrial morphofunctional defects. This USP28-PPARα-Mfn2 axis was cardioprotective in diabetic hearts. Co-IP/mass spectrometry, protein pull-down, ChIP sequencing, ChIP assays, site-specific ubiquitination mapping, cardiac-specific USP28 KO mice, AAV9-USP28 overexpression Circulation High 37994595
2024 DYRK2 phosphorylates USP28, promoting its ubiquitination and proteasomal degradation. Conversely, USP28 deubiquitinates DYRK2, stabilizing it and enhancing its kinase activity. The DYRK2 region 521-541 (particularly T525) plays a crucial role in USP28-mediated DYRK2 stabilization. This reciprocal regulation modulates p53 signaling and apoptotic responses to DNA damage. Phosphorylation assay, ubiquitination assay, Co-IP, kinase activity assay, site-directed mutants, genetic depletion with apoptosis readout Cell death and differentiation Medium 40858801
2024 USP28 deubiquitinates and stabilizes TRIM21; cardiomyocyte USP28 deubiquitinating TRIM21 negatively regulates Nrf2 antioxidant response, increasing oxidative stress and promoting cardiac hypertrophy. Identified by Co-IP/mass spectrometry screen. Co-IP/mass spectrometry substrate screen, Co-IP validation, USP28 cardiomyocyte-specific KO mice (Ang II and TAC models), deubiquitination assay, USP28 inhibitor (Otilonium Bromide) Theranostics Medium 39431010
2024 USP28 interacts with and extends the half-life of MAST1 protein via its deubiquitinating activity. CRISPR/Cas9 knockout of USP28 blocked MAST1-driven cisplatin resistance. CRISPR/Cas9 genome-wide screen, Co-IP, protein half-life assay (cycloheximide chase), deubiquitination assay, xenograft model Cellular and molecular life sciences : CMLS Medium 38498222
2024 USP28 inhibits K48-linked polyubiquitination of STAT3 and reverses its protein degradation in ADPKD. USP28 also directly interacts with and stabilizes c-Myc, both processes synergistically enhancing renal cystogenesis. Co-IP, ubiquitination assay (K48-specific), protein stability assay, pharmacological inhibition, in vivo cystogenesis model Biochemical pharmacology Medium 36442624
2024 The C-terminal domain of USP28 mediates PLK1-dependent binding to 53BP1, a dimerization-driven interaction necessary for mitotic stress memory, p53 stabilization, and cell cycle arrest. Cancer-associated missense mutations in this domain disrupt 53BP1 binding, impair nuclear localization, and destabilize USP28, compromising p53 stabilization. USP28 stabilizes p53 through deubiquitination. Cancer mutation analysis, domain interaction mapping, nuclear localization assays, deubiquitination assay (p53 substrate), Co-IP, cell cycle assays Nature communications High 41365927
2024 USP28 interacts with KRT1 and exerts deubiquitination on KRT1, maintaining KRT1 stability. USP28 knockdown results in decreased IFITM3 expression. IP-MS analysis, Co-IP, immunofluorescence, ubiquitination assay, xenograft model Experimental cell research Low 40222446
2024 USP28 stabilizes NOTCH1 intracellular domain (NICD) through direct interaction, independently of FBXW7 and the NICD-PEST domain, enhancing NOTCH1 signaling in CLL. USP28 is deleted in 90% of del(11q) CLL patients, resulting in decreased expression and dysregulation of NOTCH1 target genes. Co-IP, RNA-Seq of USP28 WT/KO cells, RBPJ ChIP, pharmacological inhibition (AZ1), primary CLL cells Leukemia Medium 40456839
2024 Crystal structure of USP28 in complex with inhibitors (AZ1, Vismodegib, FT206) reveals a common inhibitor-binding pocket. A key glutamate residue (E366 in USP28/E373 in USP25) plays a central structural role for pocket stability, inhibition, and activity. Crystal structure determination, mutagenesis of E366, inhibitor binding assays EMBO reports High 38816515
2025 DTX3L (E3 ubiquitin ligase) ubiquitinates USP28, leading to its proteasomal degradation, while USP28 counteracts by deubiquitinating both itself (autoubiquitination reversal) and DTX3L. This cross-regulation fine-tunes DSB repair in NHEJ, HR, SSA, and MMEJ pathways. Co-IP, ubiquitination assay, DSB repair pathway assays (NHEJ, HR, SSA, MMEJ), knockdown rescue experiments iScience Medium 40703443
2025 USP28 deubiquitinates SIRT1, stabilizing it and promoting osimertinib resistance in NSCLC. Co-IP confirmed the USP28-SIRT1 interaction; SIRT1 knockdown attenuated USP28 overexpression effects. Co-IP, SIRT1 protein stability assay, knockdown/overexpression rescue experiments, in vivo xenograft The Kaohsiung journal of medical sciences Low 40855785
2025 USP28 deubiquitinates and stabilizes SOX9 protein, and this is counteracted by FBXW7-mediated ubiquitination during olaparib treatment. USP28-stabilized SOX9 binds to promoters of DDR genes (SMARCA4, UIMC1, SLX4), promoting PARP inhibitor resistance in ovarian cancer. Co-IP, deubiquitination assay, ChIP-Seq, USP28 inhibitor (AZ1), FBXW7 interaction studies Cell death & disease Medium 40240356
2025 USP28 directly binds to PPARγ, preventing its ubiquitination and degradation, maintaining PPARγ signaling. In the absence of USP28, PPARγ is downregulated and its signaling pathway is inhibited, enhancing cellular defenses against excess fat; USP28 inactivation significantly reduced MASH severity in mice. Co-IP, ubiquitination assay, liver-specific USP28 KO mice (MCD diet and WD-CCl4 models), pharmacological inhibition Molecular therapy : the journal of the American Society of Gene Therapy Medium 39905730
2025 USP28 inhibitor CAS-010 disrupts the 53BP1-USP28 interaction, and USP28 catalytic activity contributes to maintaining this interaction. USP28 negatively regulates p53 transactivation in wild-type settings through its deubiquitinase activity. Crystal structure of USP28 with inhibitor, 53BP1-USP28 interaction assay, p53 transactivation assay, catalytic mutant analysis Cell chemical biology Medium 40902594
2025 USP28 interacts with HEY1 and deubiquitinates its K87 residue, stabilizing HEY1 and enhancing cancer stem-like properties of liver cancer cells. IKKβ phosphorylates HEY1 at S40, facilitating its interaction with USP28. Co-IP, site-specific deubiquitination (K87), IKKβ phosphorylation assay, in vivo tumor model, anti-PD-1 combination assay Advanced science (Weinheim, Baden-Wurttemberg, Germany) Medium 42189121
2025 USP28 directly interacts with BCR-ABL and promotes the expression of IFITM3 in CML cells, contributing to imatinib resistance. Combined inhibition of USP28 and BCR-ABL reduced IFITM3 expression and tumor progression. Co-IP (USP28-BCR-ABL interaction), overexpression, small molecule inhibitor combination, xenograft model Biomedicine & pharmacotherapy Low 38394852
2001 USP28 was identified as a new ubiquitin-specific protease gene at chromosome 11q23 with preferential expression in heart and muscle. USP28 and USP25 form a new subfamily of deubiquitinating enzymes. Both genes have alternatively spliced exons that could generate protein isoforms with distinct tissue-specific activity. Database homology search, cDNA library screening, RT-PCR, isoform characterization Genome biology Medium 11597335

Source papers

Stage 0 corpus · 84 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2007 The ubiquitin-specific protease USP28 is required for MYC stability. Nature cell biology 398 17558397
2006 A role for the deubiquitinating enzyme USP28 in control of the DNA-damage response. Cell 292 16901786
2021 Exosomal miR-500a-5p derived from cancer-associated fibroblasts promotes breast cancer cell proliferation and metastasis through targeting USP28. Theranostics 195 33664871
2016 A USP28-53BP1-p53-p21 signaling axis arrests growth after centrosome loss or prolonged mitosis. The Journal of cell biology 187 27432896
2016 53BP1 and USP28 mediate p53 activation and G1 arrest after centrosome loss or extended mitotic duration. The Journal of cell biology 187 27432897
2011 GSK-3β regulates cell growth, migration, and angiogenesis via Fbw7 and USP28-dependent degradation of HIF-1α. Blood 174 22144179
2016 53BP1 and USP28 mediate p53-dependent cell cycle arrest in response to centrosome loss and prolonged mitosis. eLife 166 27371829
2013 The deubiquitinase USP28 stabilizes LSD1 and confers stem-cell-like traits to breast cancer cells. Cell reports 146 24075993
2014 The deubiquitinase USP28 controls intestinal homeostasis and promotes colorectal cancer. The Journal of clinical investigation 129 24960159
2023 USP28 Serves as a Key Suppressor of Mitochondrial Morphofunctional Defects and Cardiac Dysfunction in the Diabetic Heart. Circulation 108 37994595
2007 Fbw7 and Usp28 regulate myc protein stability in response to DNA damage. Cell cycle (Georgetown, Tex.) 104 17873522
2018 Targeting deubiquitinase USP28 for cancer therapy. Cell death & disease 85 29415985
2014 Dual regulation of Fbw7 function and oncogenic transformation by Usp28. Cell reports 78 25437563
2020 Cancer Cells Employ Nuclear Caspase-8 to Overcome the p53-Dependent G2/M Checkpoint through Cleavage of USP28. Molecular cell 66 31982308
2015 Overexpression of deubiquitinating enzyme USP28 promoted non-small cell lung cancer growth. Journal of cellular and molecular medicine 62 25656529
2019 Distinct USP25 and USP28 Oligomerization States Regulate Deubiquitinating Activity. Molecular cell 58 30926242
2015 Usp28 counteracts Fbw7 in intestinal homeostasis and cancer. Cancer research 58 25716680
2021 USP28 facilitates pancreatic cancer progression through activation of Wnt/β-catenin pathway via stabilising FOXM1. Cell death & disease 56 34584067
2001 Characterization of alternatively spliced products and tissue-specific isoforms of USP28 and USP25. Genome biology 55 11597335
2020 Centrosome defects cause microcephaly by activating the 53BP1-USP28-TP53 mitotic surveillance pathway. The EMBO journal 54 33226141
2020 Maintaining protein stability of ∆Np63 via USP28 is required by squamous cancer cells. EMBO molecular medicine 53 32128997
2014 USP28 is recruited to sites of DNA damage by the tandem BRCT domains of 53BP1 but plays a minor role in double-strand break metabolism. Molecular and cellular biology 53 24687851
2019 Differential Oligomerization of the Deubiquitinases USP25 and USP28 Regulates Their Activities. Molecular cell 52 30926243
2021 USP28 deletion and small-molecule inhibition destabilizes c-MYC and elicits regression of squamous cell lung carcinoma. eLife 48 34636321
2018 Loss of USP28-mediated BRAF degradation drives resistance to RAF cancer therapies. The Journal of experimental medicine 45 29880484
2019 Discovery of [1,2,3]triazolo[4,5-d]pyrimidine derivatives as highly potent, selective, and cellularly active USP28 inhibitors. Acta pharmaceutica Sinica. B 44 32963944
2021 USP28: Oncogene or Tumor Suppressor? A Unifying Paradigm for Squamous Cell Carcinoma. Cells 38 34685632
2020 USP28 and USP25 are downregulated by Vismodegib in vitro and in colorectal cancer cell lines. The FEBS journal 32 32578360
2021 Inhibition of USP28 overcomes Cisplatin-resistance of squamous tumors by suppression of the Fanconi anemia pathway. Cell death and differentiation 31 34611298
2018 Knockdown of USP28 enhances the radiosensitivity of esophageal cancer cells via the c-Myc/hypoxia-inducible factor-1 alpha pathway. Journal of cellular biochemistry 29 30206969
2014 Regulation of USP28 deubiquitinating activity by SUMO conjugation. The Journal of biological chemistry 29 25359778
2023 USP28 controls SREBP2 and the mevalonate pathway to drive tumour growth in squamous cancer. Cell death and differentiation 28 37202505
2018 USP28 Deficiency Promotes Breast and Liver Carcinogenesis as well as Tumor Angiogenesis in a HIF-independent Manner. Molecular cancer research : MCR 27 29545478
2018 The HNF-1β-USP28-Claspin pathway upregulates DNA damage-induced Chk1 activation in ovarian clear cell carcinoma. Oncotarget 27 29707125
2018 USP28 contributes to the proliferation and metastasis of gastric cancer. Journal of cellular biochemistry 26 30485491
2018 The stability and oncogenic function of LIN28A are regulated by USP28. Biochimica et biophysica acta. Molecular basis of disease 26 30543854
2019 SENP1-mediated deSUMOylation of USP28 regulated HIF-1α accumulation and activation during hypoxia response. Cancer cell international 25 30622440
2018 Reverse the down regulation of miR-92b-3p by hypoxia can suppress the proliferation of pulmonary artery smooth muscle cells by targeting USP28. Biochemical and biophysical research communications 24 30149918
2019 USP28 regulates deubiquitination of histone H2A and cell proliferation. Experimental cell research 19 30910399
2024 Cardiomyocyte-derived USP28 negatively regulates antioxidant response and promotes cardiac hypertrophy via deubiquitinating TRIM21. Theranostics 18 39431010
2024 USP28 promotes tumorigenesis and cisplatin resistance by deubiquitinating MAST1 protein in cancer cells. Cellular and molecular life sciences : CMLS 15 38498222
2021 Loss of USP28 and SPINT2 expression promotes cancer cell survival after whole genome doubling. Cellular oncology (Dordrecht, Netherlands) 14 34962618
2020 Deubiquitinase USP28 inhibits ubiquitin ligase KLHL2-mediated uridine-cytidine kinase 1 degradation and confers sensitivity to 5'-azacytidine-resistant human leukemia cells. Theranostics 14 31938050
2025 USP28-Based Deubiquitinase-Targeting Chimeras for Cancer Treatment. Journal of the American Chemical Society 13 40214647
2023 Resistance to vincristine in DLBCL by disruption of p53-induced cell cycle arrest and apoptosis mediated by KIF18B and USP28. British journal of haematology 13 37190875
2022 USP28 enables oncogenic transformation of respiratory cells, and its inhibition potentiates molecular therapy targeting mutant EGFR, BRAF and PI3K. Molecular oncology 13 35364627
2021 The deubiquitinase USP28 stabilizes the expression of RecQ family helicases and maintains the viability of triple negative breast cancer cells. The Journal of biological chemistry 13 34822842
2024 The dimeric deubiquitinase USP28 integrates 53BP1 and MYC functions to limit DNA damage. Nucleic acids research 12 38227944
2024 Structural basis for the bi-specificity of USP25 and USP28 inhibitors. EMBO reports 12 38816515
2022 Stabilization of PIM Kinases in Hypoxia Is Mediated by the Deubiquitinase USP28. Cells 11 35326457
2021 USP28 promotes aerobic glycolysis of colorectal cancer by increasing stability of FOXC1. Acta biochimica Polonica 11 34106567
2023 Structure-based discovery of potent USP28 inhibitors derived from Vismodegib. European journal of medicinal chemistry 10 37075624
2024 Deubiquitinating enzyme USP28 inhibitor AZ1 alone and in combination with cisplatin for the treatment of non-small cell lung cancer. Apoptosis : an international journal on programmed cell death 9 39222275
2022 Inhibition of deubiquitinase USP28 attenuates cyst growth in autosomal dominant polycystic kidney disease. Biochemical pharmacology 9 36442624
2025 USP28 promotes PARP inhibitor resistance by enhancing SOX9-mediated DNA damage repair in ovarian cancer. Cell death & disease 8 40240356
2024 USP28 promotes tumor progression and glycolysis by stabilizing PKM2/Hif1-α in cholangiocarcinoma. Cellular oncology (Dordrecht, Netherlands) 8 39419941
2025 The deubiquitinase USP28 maintains the expression of PPARγ and its inactivation protects mice from diet-induced MASH and hepatocarcinoma. Molecular therapy : the journal of the American Society of Gene Therapy 7 39905730
2023 The deubiquitinase USP28 maintains the expression of the transcription factor MYCN and is essential in neuroblastoma cells. The Journal of biological chemistry 7 37230388
2019 The concomitant use of USP28 and p53 to predict the progression of urothelial carcinoma of the bladder. Pathology, research and practice 7 31822365
2023 Otilonium Bromide acts as a selective USP28 inhibitor and exhibits cytotoxic activity against multiple human cancer cell lines. Biochemical pharmacology 6 37579857
2022 Identification of the deubiquitinase USP28 as a novel molecular therapeutic target of ovarian cancer. Biochemical and biophysical research communications 6 36462492
2025 NOTCH1 signaling is dysregulated by loss of the deubiquitinase USP28 with del(11q), uncovering USP28 inhibition as novel therapeutic target in CLL. Leukemia 4 40456839
2025 The E3 ubiquitin ligase DTX3L and the deubiquitinase USP28 fine-tune DNA repair through mutual regulation of their protein levels. iScience 4 40703443
2024 Preclinical testing of CT1113, a novel USP28 inhibitor, for the treatment of T-cell acute lymphoblastic leukaemia. British journal of haematology 4 38685813
2024 Mitochondria-targeted catalase induced cell malignant transformation by the downregulation of p53 protein stability via USP28/miR-200b/PP2A-Cα axis. Archives of biochemistry and biophysics 4 38844154
2020 Computational approach to target USP28 for regulating Myc. Computational biology and chemistry 4 32028107
2025 Deubiquitinase USP28 promotes the malignant progression and radio-resistance of hepatocellular carcinoma by stabilizing WDHD1. Naunyn-Schmiedeberg's archives of pharmacology 3 39928151
2025 Cancer-associated USP28 missense mutations disrupt 53BP1 interaction and p53 stabilization. Nature communications 3 41365927
2024 USP28 protects development of inflammation in mouse intestine by regulating STAT5 phosphorylation and IL22 production in T lymphocytes. Frontiers in immunology 3 39076972
2023 Inhibition of Pard3 promotes breast cancer metastasis via the USP28 mediated deubiquitination of Snail1. Heliyon 3 38076191
2025 USP28-mediated deubiquitination of FOXK1 activates the Hippo signaling pathway to regulate cell proliferation and radiosensitivity in lung cancer. Life sciences 2 39983825
2025 A novel feedback loop between DYRK2 and USP28 regulates cancer homeostasis and DNA damage signaling. Cell death and differentiation 2 40858801
2025 USP28 knockdown and small molecule inhibitors promote KRT1 destabilization and sensitize hepatocellular carcinoma cells to sorafenib. Experimental cell research 1 40222446
2025 Decidual stromal cells-derived exosomes incured insufficient migration and invasion of trophoblast because of abnormal ubiquitination and degradation of Snail mediated by miR-92b-3p/USP28. BMC biology 1 40696416
2025 USP28 Promotes Osimertinib Resistance in H1975 NSCLC Cells by Deubiquitinating and Stabilizing SIRT1. The Kaohsiung journal of medical sciences 1 40855785
2025 USP28 participates in high glucose-mediated endothelial dysfunction via deubiquitinating SIRT1 protein in diabetic foot ulcers. Journal of orthopaedic surgery and research 1 40993725
2024 Investigation of the mechanism of USP28-mediated IFITM3 elevation in BCR-ABL-dependent imatinib resistance in CML. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 1 38394852
2013 ¹H, ¹³C and ¹⁵N backbone and side-chain resonance assignments of the N-terminal ubiquitin-binding domains of the human deubiquitinase Usp28. Biomolecular NMR assignments 1 23754699
2026 Lnc-USP28-6/ZBTB16 axis orchestrates NLRP3 inflammasome activation and α-synuclein SUMOylation to drive Parkinson's disease pathogenesis. Brain, behavior, and immunity 0 41775328
2026 The deubiquitinase USP28 promotes esophageal squamous cell carcinoma proliferation by stabilizing ΔNp63 protein. Cellular signalling 0 41833770
2026 Fidelity-Ensuring Consistency of Mitosis is Safeguarded by the 53BP1-USP28-p53 Pathway. bioRxiv : the preprint server for biology 0 41889881
2026 IKKβ and USP28 Regulate HEY1 Stability to Promote Cancer Stemness and Immune Evasion in Hepatocellular Carcinoma. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 0 42189121
2025 Pharmacologic interrogation of USP28 cellular function in p53 signaling. Cell chemical biology 0 40902594
2025 Targeting USP28 inhibits clear cell renal cell carcinoma growth. Cellular signalling 0 41456627

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