Affinage

DUSP3

Dual specificity protein phosphatase 3 · UniProt P51452

Length
185 aa
Mass
20.5 kDa
Annotated
2026-06-09
63 papers in source corpus 27 papers cited in narrative 27 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 7/7 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

DUSP3 (VHR) is a constitutively expressed dual-specificity phosphatase that acts as a tonic brake on tyrosine-phosphorylation signaling, particularly within the MAP kinase pathways (PMID:10224087, PMID:11085983, PMID:16604064). Catalysis proceeds through a classical cysteine-based phosphatase mechanism: Cys124 is the nucleophile that forms a covalent thiol-phosphate intermediate, Asp92 serves as the general acid protonating the leaving group, and the rate-limiting step is decomposition of the phosphoenzyme intermediate via a highly dissociative transition state (PMID:7961745, PMID:8519766, PMID:8679534). A shallow active-site pocket accommodates pSer/pThr/pTyr, but structural and peptide-library analyses show a strong preference for dephosphorylating phosphotyrosine within bisphosphorylated -pTXpY- motifs, with phosphotyrosine occupying the deep catalytic cleft and phosphothreonine tethered in an adjacent basic pocket (PMID:8650541, PMID:11863439, PMID:23322772). Through this activity DUSP3 dephosphorylates and inactivates ERK1/2 and JNK1/2, and these substrates define its role in cell-cycle progression: loss of DUSP3 hyperactivates ERK and JNK, causing G1/S and G2/M arrest, senescence, and ERK-dependent multipolar mitotic spindles (PMID:10224087, PMID:11971192, PMID:16604064, PMID:27423135). The enzyme is itself regulated on multiple axes — activating Tyr138 phosphorylation by ZAP-70 and Tyk2 (required for activity toward the ERK–Elk-1 pathway and STAT5) (PMID:12447358, PMID:17785772), allosteric activation by VRK3 binding that is enhanced by nuclear HSP70 (PMID:16845380, PMID:27941812), and negative regulation by homodimerization that occludes the active site (PMID:24798147). Beyond MAP kinases, DUSP3 dephosphorylates a defined set of substrates including STAT5 (PMID:17785772), FAK and paxillin to control focal adhesion turnover and migration (PMID:28759036), occludin to maintain tight-junction barrier integrity (PMID:35705979), nucleophosmin (Y29/Y67/Y271) to fine-tune p53 stability and DNA-damage responses (PMID:33777934), and HnRNP C to regulate IRES-dependent translation of c-MYC and XIAP (PMID:38538536). At the organismal level DUSP3 is pro-angiogenic (PMID:24886454) and shapes innate immune responses, where its deficiency confers tolerance to endotoxin/septic shock through macrophage-dependent ERK1/2 regulation and M2 polarization (PMID:25876765).

Mechanistic history

Synthesis pass · year-by-year structured walk · 20 steps
  1. 1994 High

    Established the catalytic mechanism of VHR/DUSP3 by identifying Cys124 as the nucleophile, resolving how a dual-specificity phosphatase achieves both pTyr and pSer/pThr hydrolysis.

    Evidence C124S mutagenesis, 32P phosphoenzyme intermediate trapping, iodoacetate alkylation, in vitro assay

    PMID:7961745

    Open questions at the time
    • Did not identify physiological substrates
    • Subcellular context and regulation unaddressed
  2. 1995 High

    Defined the kinetic pathway, showing the rate-limiting step is breakdown of the phosphoenzyme intermediate rather than initial phosphoryl transfer.

    Evidence Pre-steady-state burst kinetics with pNPP, linear free-energy analysis

    PMID:8519766

    Open questions at the time
    • Used artificial substrate only
    • No physiological substrate kinetics
  3. 1996 High

    Provided the structural basis for dual specificity and substrate recognition through the first crystal structure and transition-state/general-acid analysis.

    Evidence X-ray crystallography at 2.1 Å; kinetic isotope effects with D92N/S131A mutagenesis

    PMID:8650541 PMID:8679534

    Open questions at the time
    • No substrate-bound complex
    • Specificity determinants for cellular substrates not validated
  4. 1999 High

    Identified ERK1/2 as authentic cellular substrates, defining DUSP3 as a constitutive nuclear brake on MAP kinase signaling.

    Evidence Substrate-trap affinity enrichment, in vitro kinetics, immunodepletion in COS-1 cells

    PMID:10224087

    Open questions at the time
    • JNK/p38 selectivity context-dependent
    • Recruitment to substrates not defined
  5. 2000 High

    Demonstrated DUSP3 tonically suppresses ERK and JNK in T cells, and that the active-site Cys is redox-sensitive, linking enzyme activity to signaling and metal-mediated regulation.

    Evidence WT/catalytically dead VHR in Jurkat cells with reporters; Cu2+/Zn2+ inactivation with iodoacetate labeling

    PMID:11051099 PMID:11085983

    Open questions at the time
    • Physiological relevance of metal inactivation in vivo unestablished
    • p38/NF-κB exclusion may be cell-type specific
  6. 2002 High

    Resolved substrate recognition and activating regulation: a substrate-bound structure explained pTyr preference within -pTXpY- motifs, JNK was confirmed as substrate, and ZAP-70-mediated Tyr138 phosphorylation was shown to be required for activity.

    Evidence C124S–bisphosphopeptide crystal structure; JNK substrate trap with c-Jun inhibition; Co-IP/kinase assay/Y138F mutant in T cells

    PMID:11863439 PMID:11971192 PMID:12447358

    Open questions at the time
    • Structural basis of Tyr138-dependent activation not solved
    • Whether c-Jun occlusion operates in vivo unclear
  7. 2006 High

    Placed DUSP3 upstream of ERK/JNK in cell-cycle control and identified VRK3 as a kinase-independent allosteric activator, revealing both phenotypic consequence and a novel activation mechanism.

    Evidence RNAi with senescence/cell-cycle assays and JNK/ERK inhibitor rescue; Co-IP and in vitro activity assay with VRK3 kinase-dead mutant

    PMID:16604064 PMID:16845380

    Open questions at the time
    • Molecular mechanism of VRK3 allosteric activation undefined
    • Direct cell-cycle substrate beyond ERK/JNK not identified
  8. 2007 High

    Extended substrate range to interferon-activated STAT5 and confirmed Tyk2-mediated Tyr138 phosphorylation as a requirement for activity, integrating DUSP3 into cytokine signaling.

    Evidence Phosphatase assay, Co-IP, VHR Y138F and STAT5 SH2-domain mutants, reporter assays

    PMID:17785772

    Open questions at the time
    • In vivo relevance to IFN responses not tested
    • Mechanism of SH2-dependent recognition not structurally defined
  9. 2008 Medium

    Linked DUSP3 subcellular distribution to cancer, showing nuclear relocalization and post-translational stabilization in cervical cancer cells.

    Evidence Immunofluorescence, Western blot, RT-PCR in keratinocytes vs cancer lines

    PMID:18505570

    Open questions at the time
    • Mechanism of stabilization unknown
    • Functional consequence of nuclear shift not directly tested
  10. 2013 Medium

    Identified the DNA-damage-induced interactome (NPM, HnRNP C1/C2, Nucleolin) and systematically mapped two substrate-peptide binding classes, broadening DUSP3 substrate logic beyond MAP kinases.

    Evidence GST pulldown with LC-MS/MS and Co-IP validation; combinatorial peptide library with mutagenesis and modeling

    PMID:23322772 PMID:24245651

    Open questions at the time
    • Pulldown interactions not yet shown to be catalytic substrates in 2013
    • Class II inverted binding mode based partly on modeling
  11. 2014 High

    Used knockout mice to establish DUSP3 as pro-angiogenic and to identify a PKC-linked, ERK-independent vascular function, demonstrating non-MAPK signaling roles in vivo.

    Evidence DUSP3-knockout mice, endothelial RNAi, Matrigel/aortic-ring/tube-formation assays, phosphoprotein analysis

    PMID:24886454

    Open questions at the time
    • Direct PKC-pathway substrate not identified
    • Mechanism connecting DUSP3 to angiogenesis incomplete
  12. 2014 High

    Revealed homodimerization as a negative autoregulatory mechanism that occludes the active site, adding a layer of intrinsic activity control.

    Evidence pBPA photo-cross-linking and chemical cross-linking identifying Phe68; activity assay of dimer vs monomer

    PMID:24798147

    Open questions at the time
    • Cellular triggers governing monomer/dimer equilibrium unknown
    • No structure of the dimer interface
  13. 2015 High

    Defined DUSP3 as an innate-immune regulator, showing macrophage-dependent control of endotoxin/septic shock tolerance through ERK1/2 and M2 polarization.

    Evidence DUSP3-/- mice in LPS/CLP models, adoptive bone-marrow transfer, FACS, ERK1/2 and TNF readouts

    PMID:25876765

    Open questions at the time
    • Direct macrophage substrate beyond ERK pathway not pinned down
    • Transcriptional drivers of M2 skewing unresolved
  14. 2016 Medium

    Connected DUSP3 to mitotic fidelity and to a neuronal protective axis: ERK-dependent control of spindle bipolarity and HSP70-enhanced phosphatase activity downstream of VRK3.

    Evidence RNAi with ERK-inhibitor/overexpression rescue and spindle imaging; Co-IP, activity assay, NLS-HSP70 overexpression in neurons

    PMID:27423135 PMID:27941812

    Open questions at the time
    • HSP70 activation mechanism (non-chaperone) not structurally defined
    • Spindle phenotype substrate(s) beyond ERK unclear
  15. 2017 High

    Identified FAK and paxillin as direct substrates, establishing DUSP3 control of focal adhesion turnover and cell migration.

    Evidence Co-IP, in vitro phosphatase assay with recombinant FAK, knockout cells, overexpression, focal adhesion microscopy

    PMID:28759036

    Open questions at the time
    • Spatial regulation of cytoplasmic DUSP3 at adhesions not defined
    • Interplay with EGFR signaling not fully resolved
  16. 2020 High

    Demonstrated long-range allosteric communication from the variable insert (Asn74) to the catalytic acid loop, revealing a structural route for activity modulation distant from the active site.

    Evidence Solution NMR, MD simulations, steady-state/rapid kinetics with N74A mutagenesis

    PMID:32348128

    Open questions at the time
    • Physiological ligand or modifier engaging this allosteric route unknown
    • Link to VRK3/HSP70 activation not established
  17. 2021 High

    Showed DUSP3 dephosphorylates NPM at three specific tyrosines to fine-tune p53 stability and the DNA-damage response, converting an earlier interactome hit into a defined catalytic substrate.

    Evidence Co-IP, site-specific phospho-antibodies, in vitro phosphatase assay, siRNA, p53 stability/transcription assays

    PMID:33777934

    Open questions at the time
    • Whether NPM dephosphorylation requires VHR Tyr138 activation untested
    • Nucleolar dynamics regulation mechanism incomplete
  18. 2022 High

    Identified occludin as a direct substrate, linking DUSP3 to tight-junction stability and epithelial barrier function via both direct OCLN dephosphorylation and indirect FAK suppression.

    Evidence Proximity ligation assay, in vitro phosphatase assay, OCLN Tyr-to-Ala mutants, ubiquitination and barrier assays in deficient cells

    PMID:35705979

    Open questions at the time
    • In vivo epithelial relevance not tested
    • Which OCLN tyrosines are primary remains to be ranked
  19. 2024 Medium

    Extended DUSP3 function to translational control, showing dephosphorylation of HnRNP C regulates IRES-dependent translation of c-MYC and XIAP and placing DUSP3 in ribosomal fractions.

    Evidence siRNA, phospho-HNRNPC immunoblot, RNA-IP, polysome fractionation, qPCR vs protein

    PMID:38538536

    Open questions at the time
    • No in vitro reconstitution of HNRNPC dephosphorylation by DUSP3
    • Direct vs indirect effect on translation not fully separated
  20. 2025 Medium

    Mapped novel allosteric ligand-binding pockets distinct from the catalytic site, providing a structural foundation for allosteric modulators.

    Evidence Fluorine NMR fragment screening with X-ray crystallography of fragment-bound VHR

    PMID:39959108

    Open questions at the time
    • Functional consequence of occupying these sites not demonstrated
    • No lead compound with cellular activity

Open questions

Synthesis pass · forward-looking unresolved questions
  • How DUSP3's many regulatory inputs (Tyr138 phosphorylation, VRK3/HSP70 binding, dimerization, allosteric insert dynamics) are integrated to select among its diverse substrates in different compartments and cell types remains unresolved.
  • No unified model of substrate selection
  • Compartment-specific regulation incompletely mapped
  • Disease-relevant substrate priorities not defined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 6 GO:0016787 hydrolase activity 3 GO:0098772 molecular function regulator activity 3
Localization
GO:0005634 nucleus 4 GO:0005829 cytosol 2 GO:0005730 nucleolus 1 GO:0005840 ribosome 1
Pathway
R-HSA-162582 Signal Transduction 4 R-HSA-1640170 Cell Cycle 2 R-HSA-168256 Immune System 1
Partners
ERK1/2FAKHNRNPCHSP70JNKNPM1STAT5VRK3

Evidence

Reading pass · 27 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1994 Cys124 is the catalytic nucleophile of VHR/DUSP3: the C124S mutation abolishes both Tyr and Thr/Ser phosphatase activities, a thiol-phosphate enzyme intermediate forms at Cys124 during catalysis, and Cys124 is specifically alkylated by iodoacetate, establishing a mechanism identical to that of tyrosine-specific PTPs. Active-site mutagenesis (C124S), 32P-labeling of phosphoenzyme intermediate, iodoacetate alkylation, in vitro phosphatase assay The Journal of biological chemistry High 7961745
1995 Pre-steady-state burst kinetics demonstrate that VHR/DUSP3 catalysis proceeds through a phosphoenzyme intermediate, and the rate-limiting step is decomposition of this intermediate (dephosphorylation), not the initial phosphoryl transfer from substrate to Cys124. Pre-steady-state burst kinetic analysis with p-nitrophenyl phosphate; linear free-energy relationship analysis Biochemistry High 8519766
1996 Crystal structure of VHR/DUSP3 at 2.1 Å resolution reveals a shallow active-site pocket (explaining dual-specificity toward pSer/pThr/pTyr) compared to the deeper pocket of tyrosine-specific PTPs, positively charged crevices near the active site, and a 'recognition region' (helix α1 to strand β1) that likely determines substrate specificity differences among DSPs and PTPs. X-ray crystallography at 2.1 Å resolution Science High 8650541
1996 Kinetic isotope effect measurements show VHR/DUSP3 employs a highly dissociative transition state for phosphoryl transfer (similar to uncatalyzed reaction and other PTPs), with Asp92 acting as general acid to protonate the leaving group; D92N mutant loses general acid assistance; S131A mutation raises the pKa of the nucleophilic Cys but does not alter transition-state structure. Kinetic isotope effects (18O nonbridge, 18O bridge, 15N) measured by isotope ratio mass spectrometry; site-directed mutagenesis (D92N, S131A, D92N/S131A) Biochemistry High 8679534
1999 ERK1 and ERK2 are authentic substrates of VHR/DUSP3: VHR specifically dephosphorylates and inactivates ERK1/2 in vitro and in vivo (but not p38 or JNK in this study), with a second-order rate constant of ~40,000 M⁻¹s⁻¹; immunodepletion of endogenous VHR eliminates cellular ERK dephosphorylation; VHR is constitutively expressed and nuclear. Covalently immobilized mutant VHR affinity trap, kinetic in vitro phosphatase assay, transfection in COS-1 cells, immunodepletion The Journal of biological chemistry High 10224087
2000 Introduction of exogenous VHR into Jurkat T cells suppresses TCR-induced activation of ERK1/2 and JNK1/2 (and NFAT/AP-1 and Elk/c-Jun-driven reporters), but not p38 or NF-κB; catalytically inactive VHR mutants cause increased gene activation, indicating that endogenous VHR tonically suppresses the ERK and JNK pathways in T cells. Transfection of wild-type and catalytically inactive VHR in Jurkat T cells; luciferase reporter assays; kinase activity assays The Journal of biological chemistry High 11085983
2000 Cu²⁺ ions potently and reversibly inactivate VHR/DUSP3 at submicromolar concentrations by oxidizing the active-site Cys124, as shown by loss of [¹⁴C]iodoacetate labeling of that residue; the reduction potential of VHR is estimated at −331 mV. Zn²⁺ inactivates VHR through a distinct mechanism not involving active-site Cys124. In vitro phosphatase activity assay with metal ions; [¹⁴C]iodoacetate active-site labeling; DTT reversal; Y78F mutant comparison Archives of biochemistry and biophysics Medium 11051099
2002 Crystal structure at 2.75 Å of catalytically inactive C124S VHR in complex with a bisphosphorylated MAP kinase activation-loop peptide reveals that phosphotyrosine occupies the deep active-site cleft while phosphothreonine is tethered in a nearby basic pocket containing Arg158, explaining VHR's strong preference for dephosphorylating pTyr over pThr within bisphosphorylated -pTXpY- substrates. X-ray crystallography of C124S VHR–bisphosphorylated peptide complex at 2.75 Å; biochemical dephosphorylation assays; mutagenesis Biochemistry High 11863439
2002 VHR/DUSP3 efficiently dephosphorylates and inactivates JNK; the catalytically inactive C124S VHR forms a tight complex with activated JNK in vivo (substrate trap); c-Jun inhibits VHR-mediated JNK dephosphorylation in vitro by sterically blocking phosphorylation-site access when JNK and c-Jun are complexed, but c-Jun does not affect VHR activity toward ERK or artificial substrates. In vitro phosphatase assay, C124S substrate trap in vivo, c-Jun inhibition assay in vitro Oncogene High 11971192
2002 VHR/DUSP3 is phosphorylated at Tyr138 by the ZAP-70 tyrosine kinase following TCR stimulation; Tyr138 phosphorylation is required for VHR to inhibit the ERK2-Elk-1 pathway, and the VHR(Y138F) mutant augments TCR-induced ERK2 activity and IL-2 gene activation. Co-immunoprecipitation, in vitro kinase assay, site-directed mutagenesis (Y138F), transfection/reporter assay in T cells Nature immunology High 12447358
2006 RNAi-mediated loss of VHR/DUSP3 causes cell-cycle arrest at G1/S and G2/M transitions, senescence markers (β-galactosidase staining, autophagosomes, p21Cip/Waf1 upregulation, decreased telomerase), and hyperactivation of JNK and ERK; the cell-cycle arrest is reversed by JNK and ERK inhibition or knockdown, placing VHR upstream of these MAP kinases in cell-cycle regulation. RNAi knockdown, cell-cycle analysis, senescence assays (β-gal, autophagosome), kinase activity assays, JNK/ERK inhibitor rescue Nature cell biology High 16604064
2006 VRK3 kinase binds directly to VHR/DUSP3 and enhances its phosphatase activity toward ERK in the nucleus through a mechanism independent of VRK3 kinase activity, thereby suppressing ERK signaling; VRK3 defines a class of 'phosphatase-activating kinases'. Co-immunoprecipitation, in vitro phosphatase activity assay with VRK3, VRK3 kinase-dead mutant, nuclear co-localization studies Nature cell biology High 16845380
2007 VHR/DUSP3 selectively dephosphorylates IFN-α/β-activated, tyrosine-phosphorylated STAT5 in the nucleus, inhibiting STAT5 transcriptional activity; Tyr138 phosphorylation of VHR is required for its phosphatase activity toward STAT5; the Src homology 2 (SH2) domain of STAT5 is required for effective dephosphorylation by VHR; Tyk2 kinase phosphorylates VHR at Tyr138. Phosphatase assay, co-immunoprecipitation, VHR Y138F mutant, STAT5 SH2-domain mutant analysis, reporter assays Journal of immunology High 17785772
2008 VHR/DUSP3 localizes to the cytoplasm in primary keratinocytes but relocates to both cytoplasm and nucleus in cervical cancer cell lines; this relocalization correlates with post-translational stabilization of VHR protein (not increased DUSP3 transcription) in cancer cells. Immunofluorescence/confocal microscopy, Western blotting, RT-PCR BMC cancer Medium 18505570
2013 GST-DUSP3 pulldown from HeLa cells exposed to gamma/UV radiation followed by LC-MS/MS identified Nucleophosmin (NPM), HnRNP C1/C2, and Nucleolin as direct DUSP3-interacting proteins; these interactions were validated by co-immunoprecipitation. GST pulldown, LC-MS/MS proteomics, co-immunoprecipitation validation Journal of proteome research Medium 24245651
2013 Systematic combinatorial peptide library profiling reveals VHR/DUSP3 recognizes two distinct classes of phosphotyrosine peptide substrates: Class I ((D/E/ϕ)(D/S/N/T/E)(P/I/M/S/A/V)pY(G/A/S/Q)) binding in canonical orientation, and Class II ((V/A)P(I/L/M/V/F)Xn pY) binding in an inverted orientation where the N-terminus interacts with Asp164; this alternative binding mode was supported by site-directed mutagenesis and molecular modeling. Combinatorial peptide library screening, site-directed mutagenesis, molecular modeling The Journal of biological chemistry High 23322772
2014 VHR/DUSP3 can homodimerize inside cells; photo-cross-linking (pBPA amber suppression) and chemical cross-linkers identified Phe68 as a residue involved in dimerization; dimerization reduces VHR catalytic activity, suggesting the dimer interface occludes the active site as a negative regulatory mechanism. Unnatural amino acid (pBPA) photo-cross-linking, chemical cross-linking, in vitro phosphatase activity assay of dimeric vs monomeric VHR ACS chemical biology High 24798147
2014 DUSP3 deficiency in mice reduces neo-vascularization (b-FGF Matrigel plug, LLC tumor xenograft, aortic ring assay); DUSP3 knockdown in human endothelial cells reduces tube formation and spheroid sprouting, associated with increased PKC phosphorylation (not altered ERK1/2, JNK1/2, or EGFR phosphorylation in this context), identifying DUSP3 as a pro-angiogenic phosphatase. DUSP3-knockout mice (homologous recombination), RNAi in endothelial cells, Matrigel tube formation, aortic ring assay, phosphoprotein analysis Molecular cancer High 24886454
2015 DUSP3 deficiency in mice promotes tolerance to LPS-induced endotoxin shock and polymicrobial septic shock; this is macrophage-dependent and transferable by adoptive transfer; DUSP3-/- mice show increased M2-like macrophages, decreased TNF production, and impaired ERK1/2 activation, demonstrating DUSP3 regulates innate immune responses through ERK1/2 and macrophage polarization. DUSP3-/- mice, LPS/CLP models, adoptive bone-marrow transfer, FACS macrophage phenotyping, ERK1/2 phosphorylation assay, ELISA for TNF Journal of immunology High 25876765
2016 Nuclear HSP70 enhances VHR/DUSP3 phosphatase activity via direct protein–protein interaction (not chaperone activity); VRK3 facilitates nuclear localization of HSP70; this VRK3/HSP70/VHR axis suppresses excessive ERK activation following glutamate excitotoxicity and reduces apoptosis and Aβ accumulation. Co-immunoprecipitation, in vitro phosphatase activity assay, nuclear localization signal-fused HSP70 overexpression, VRK3-deficient neurons, VRK3/HSP70 knockdown Scientific reports Medium 27941812
2016 Loss of Dusp3/VHR by RNAi in human mitotic cancer cells causes multipolar spindle formation in an ERK1/2-dependent manner; normal bipolar spindle structure is restored by chemical inhibition of ERK1/2 or ectopic Dusp3 overexpression, placing Dusp3 upstream of ERK1/2 in mitotic spindle regulation. RNAi knockdown, ERK1/2 inhibitor rescue, Dusp3 overexpression rescue, spindle morphology analysis by microscopy FEBS letters Medium 27423135
2017 VHR/DUSP3 directly dephosphorylates FAK (focal adhesion kinase) and paxillin in vitro; VHR overexpression decreases FAK tyrosine phosphorylation and VHR deficiency increases it; VHR-knockout cells show stronger FAK and paxillin phosphorylation, higher EGFR phosphorylation, long-lasting trailing ends, slower focal adhesion turnover, and elevated cell migration, identifying VHR as a FAK phosphatase regulating focal adhesion dynamics. Co-immunoprecipitation, in vitro phosphatase assay with recombinant VHR and FAK, VHR-knockout mouse-derived cells, VHR overexpression, focal adhesion microscopy Oncogene High 28759036
2020 Allosteric communication exists between the variable insert region (residue Asn74) and the catalytic acid loop of VHR/DUSP3: N74A mutation rigidifies the acid loop (NMR and MD simulations), disrupts active-site hydrogen bonds, weakens substrate affinity, and reduces substrate cleavage and hydrolysis rates >2-fold, despite the variable insert being ~20 Å from the active site. Solution NMR, molecular dynamics simulations, steady-state and rapid kinetic measurements, N74A site-directed mutagenesis Biochemistry High 32348128
2021 DUSP3 directly dephosphorylates Nucleophosmin (NPM) at tyrosines Y29, Y67, and Y271 after UV-radiation stress; DUSP3 knockdown causes early nucleolus exit of NPM and ARF, disruption of the HDM2-p53 interaction, increased p53-Ser15 phosphorylation, prolonged p53 half-life, and enhanced p53 transcriptional activity, demonstrating that DUSP3-mediated NPM dephosphorylation fine-tunes p53 signaling to maintain genomic stability. Co-immunoprecipitation, phospho-specific antibodies to individual NPM Tyr residues, in vitro phosphatase assay, DUSP3 siRNA knockdown, p53 stability/transcription assays Frontiers in cell and developmental biology High 33777934
2022 DUSP3 directly dephosphorylates occludin (OCLN) tyrosine residues; DUSP3 deficiency increases OCLN tyrosine phosphorylation, OCLN ubiquitination, and OCLN degradation, leading to disrupted tight junction ZO-1 distribution and impaired epithelial barrier function; DUSP3 also suppresses OCLN kinase FAK to reduce OCLN phosphorylation indirectly. Proximity ligation assay, immunoblotting, in vitro phosphatase assay, DUSP3-deficient cells, OCLN Tyr-to-Ala mutants, ubiquitination assay Journal of biomedical science High 35705979
2024 DUSP3 dephosphorylates HnRNP C (HNRNPC) tyrosine residues; DUSP3 knockdown causes tyrosine hyperphosphorylation of HNRNPC, increasing its RNA-binding ability and its association with IRES trans-acting factor (ITAF) complexes, leading to enhanced IRES-dependent translation of c-MYC and XIAP mRNAs. DUSP3 is present in 40S, monosome, and polysome fractions interacting with HNRNPC, PABP, and Nucleolin. DUSP3 siRNA knockdown, tyrosine phospho-HNRNPC immunoblotting, RNA immunoprecipitation, polysome fractionation, qPCR for mRNA levels vs protein levels Biology of the cell Medium 38538536
2025 Fragment-based screening using fluorine NMR identified novel ligand-binding sites on VHR/DUSP3 distant from the conserved active site (allosteric sites); crystal structures confirmed fragment–protein interactions at these novel sites, providing structural basis for allosteric modulation. Fluorine NMR fragment screening, X-ray crystallography of fragment-bound VHR ACS omega Medium 39959108

Source papers

Stage 0 corpus · 63 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1996 Crystal structure of the dual specificity protein phosphatase VHR. Science (New York, N.Y.) 294 8650541
1999 Extracellular regulated kinases (ERK) 1 and ERK2 are authentic substrates for the dual-specificity protein-tyrosine phosphatase VHR. A novel role in down-regulating the ERK pathway. The Journal of biological chemistry 142 10224087
2000 Inhibitory role for dual specificity phosphatase VHR in T cell antigen receptor and CD28-induced Erk and Jnk activation. The Journal of biological chemistry 126 11085983
1994 The catalytic role of Cys124 in the dual specificity phosphatase VHR. The Journal of biological chemistry 107 7961745
2006 Loss of the VHR dual-specific phosphatase causes cell-cycle arrest and senescence. Nature cell biology 106 16604064
2000 Effects of metal ions on the activity of protein tyrosine phosphatase VHR: highly potent and reversible oxidative inactivation by Cu2+ ion. Archives of biochemistry and biophysics 90 11051099
2002 Tyrosine phosphorylation of VHR phosphatase by ZAP-70. Nature immunology 82 12447358
2002 Structural basis for the recognition of a bisphosphorylated MAP kinase peptide by human VHR protein Phosphatase. Biochemistry 78 11863439
2006 Negative regulation of ERK activity by VRK3-mediated activation of VHR phosphatase. Nature cell biology 69 16845380
2001 Inhibition of T cell antigen receptor signaling by VHR-related MKPX (VHX), a new dual specificity phosphatase related to VH1 related (VHR). The Journal of biological chemistry 69 11733513
2001 Synthesis of a tetronic acid library focused on inhibitors of tyrosine and dual-specificity protein phosphatases and its evaluation regarding VHR and cdc25B inhibition. Journal of medicinal chemistry 66 11563920
1996 Transition-state structures for the native dual-specific phosphatase VHR and D92N and S131A mutants. Contributions to the driving force for catalysis. Biochemistry 57 8679534
2002 Dual-specificity protein tyrosine phosphatase VHR down-regulates c-Jun N-terminal kinase (JNK). Oncogene 54 11971192
2008 Cervix carcinoma is associated with an up-regulation and nuclear localization of the dual-specificity protein phosphatase VHR. BMC cancer 51 18505570
2007 Cutting edge: selective tyrosine dephosphorylation of interferon-activated nuclear STAT5 by the VHR phosphatase. Journal of immunology (Baltimore, Md. : 1950) 50 17785772
2009 Multidentate small-molecule inhibitors of vaccinia H1-related (VHR) phosphatase decrease proliferation of cervix cancer cells. Journal of medicinal chemistry 47 19888758
2014 DUSP3/VHR is a pro-angiogenic atypical dual-specificity phosphatase. Molecular cancer 44 24886454
2007 BRCA1-IRIS activates cyclin D1 expression in breast cancer cells by downregulating the JNK phosphatase DUSP3/VHR. International journal of cancer 44 17278098
1995 Transition state and rate-limiting step of the reaction catalyzed by the human dual-specificity phosphatase, VHR. Biochemistry 43 8519766
2015 DUSP3 Genetic Deletion Confers M2-like Macrophage-Dependent Tolerance to Septic Shock. Journal of immunology (Baltimore, Md. : 1950) 41 25876765
2015 VHR/DUSP3 phosphatase: structure, function and regulation. The FEBS journal 35 25757426
1996 VHR and PTP1 protein phosphatases exhibit remarkably different active site specificities toward low molecular weight nonpeptidic substrates. Biochemistry 35 8755712
2002 4-isoavenaciolide covalently binds and inhibits VHR, a dual-specificity phosphatase. FEBS letters 34 12163160
2006 Regulation of MAP kinases by the VHR dual-specific phosphatase: implications for cell growth and differentiation. Cell cycle (Georgetown, Tex.) 33 17012840
2014 Dusp3 and Psme3 are associated with murine susceptibility to Staphylococcus aureus infection and human sepsis. PLoS pathogens 28 24901344
2016 VRK3-mediated nuclear localization of HSP70 prevents glutamate excitotoxicity-induced apoptosis and Aβ accumulation via enhancement of ERK phosphatase VHR activity. Scientific reports 25 27941812
2017 Deficiency in VHR/DUSP3, a suppressor of focal adhesion kinase, reveals its role in regulating cell adhesion and migration. Oncogene 24 28759036
2019 DUSP3/VHR: A Druggable Dual Phosphatase for Human Diseases. Reviews of physiology, biochemistry and pharmacology 22 30069819
2007 Identification of a potent inhibitor of human dual-specific phosphatase, VHR, from computer-aided and NMR-based screening to cellular effects. Chembiochem : a European journal of chemical biology 22 17933004
2013 Proteomic, cellular, and network analyses reveal new DUSP3 interactions with nucleolar proteins in HeLa cells. Journal of proteome research 20 24245651
2017 Dusp3 deletion in mice promotes experimental lung tumour metastasis in a macrophage dependent manner. PloS one 19 29020102
1995 Human dual specificity phosphatase VHR activates maturation promotion factor and triggers meiotic maturation in Xenopus oocytes. The Journal of biological chemistry 19 7775484
2018 Revisiting the roles of VHR/DUSP3 phosphatase in human diseases. Clinics (Sao Paulo, Brazil) 15 30208163
2008 High intracellular Zn2+ ions modulate the VHR, ZAP-70 and ERK activities of LNCaP prostate cancer cells. Cellular & molecular biology letters 15 18311544
2020 DUSP3 maintains genomic stability and cell proliferation by modulating NER pathway and cell cycle regulatory proteins. Cell cycle (Georgetown, Tex.) 14 32380926
2004 A new VHR dual-specificity protein tyrosine phosphatase inhibitor from Dendrobium moniliforme. Planta medica 14 15386195
2024 Gastric cancer cell-derived exosomal miR-541-5p induces M2 macrophage polarization through DUSP3/JAK2/STAT3 pathway. BMC cancer 13 39103776
2021 Nucleophosmin Protein Dephosphorylation by DUSP3 Is a Fine-Tuning Regulator of p53 Signaling to Maintain Genomic Stability. Frontiers in cell and developmental biology 13 33777934
2017 Loss of DUSP3 activity radiosensitizes human tumor cell lines via attenuation of DNA repair pathways. Biochimica et biophysica acta. General subjects 13 28389334
2002 Inhibition of VHR dual-specificity protein tyrosine phosphatase activity by flavonoids isolated from Scutellaria baicalensis: structure-activity relationships. Planta medica 13 12494330
2021 The genetic deletion of the Dual Specificity Phosphatase 3 (DUSP3) attenuates kidney damage and inflammation following ischaemia/reperfusion injury in mouse. Acta physiologica (Oxford, England) 12 34704357
2014 Unnatural amino acid mutagenesis reveals dimerization as a negative regulatory mechanism of VHR's phosphatase activity. ACS chemical biology 12 24798147
2024 Dehydroandrographolide facilitates M2 macrophage polarization by downregulating DUSP3 to inhibit sepsis-associated acute kidney injury. Immunity, inflammation and disease 11 38629726
2013 Specificity profiling of dual specificity phosphatase vaccinia VH1-related (VHR) reveals two distinct substrate binding modes. The Journal of biological chemistry 11 23322772
1994 Localization of the VHR phosphatase gene and its analysis as a candidate for BRCA1. Genomics 11 7829067
2022 DUSP3 regulates phosphorylation-mediated degradation of occludin and is required for maintaining epithelial tight junction. Journal of biomedical science 10 35705979
2010 A context-sensitive technique robust to registration noise for change detection in VHR multispectral images. IEEE transactions on image processing : a publication of the IEEE Signal Processing Society 8 20215070
2025 Fragment Screening Identifies Novel Allosteric Binders and Binding Sites in the VHR (DUSP3) Phosphatase. ACS omega 7 39959108
2016 Reduced levels of Dusp3/Vhr phosphatase impair normal spindle bipolarity in an Erk1/2 activity-dependent manner. FEBS letters 7 27423135
2023 Dual Specificity Phosphatase 3 (DUSP3) Knockdown Alleviates Acute Myocardial Infarction Damage via Inhibiting Apoptosis and Inflammation. Current neurovascular research 6 37272465
2020 Allosteric Impact of the Variable Insert Loop in Vaccinia H1-Related (VHR) Phosphatase. Biochemistry 6 32348128
2024 Exercise training-driven exosomal miRNA-323-5p activity suppresses adipogenic conversion of 3T3-L1 cells via the DUSP3/ERK pathway. Biochemical and biophysical research communications 5 39083976
2014 RE12 derivatives displaying Vaccinia H1-related phosphatase (VHR) inhibition in the presence of detergent and their anti-proliferative activity against HeLa cells. Bioorganic & medicinal chemistry 5 24702858
2024 l-Methionine potentiates anticancer activity of Sorafenib by epigenetically altering DUSP3/ERK pathway in hepatocellular carcinoma. Journal of biochemical and molecular toxicology 4 38367245
2021 UV Radiation-induced Impairment of Cellular Morphology and Motility is Enhanced by DUSP3/VHR Loss and FAK Activation. Cell biochemistry and biophysics 4 33479884
2024 DUSP3 modulates IRES-dependent translation of mRNAs through dephosphorylation of the HNRNPC protein in cells under genotoxic stimulus. Biology of the cell 3 38538536
2024 The Dual-specificity Phosphatase 3 (DUSP3): A Potential Target Against Renal Ischemia/Reperfusion Injury. Transplantation 3 39466786
2025 DUSP3 restrains the progression and stemness property of osteosarcoma through regulating EGFR/STAT3/SOX2 axis. International journal of biological sciences 2 39744427
2025 USC-Derived Small Extracellular Vesicles-Functionalized Scaffolds Promote Scarless Vaginal Defect Repair via Delivery of Decorin and DUSP3 Proteins. International journal of nanomedicine 1 39931530
2025 Targeting DUSP3 promotes cell senescence by activating the notch1 pathway to treat hepatocellular carcinoma. Tissue & cell 1 39954561
2025 BCL6, DUSP3, and IL6R Are Identified as Shared Druggable Immune-Regulatory Axis in Atrial Fibrillation and Atherosclerosis Through Integrative In Silico and In Vitro Analysis. Human mutation 1 41262879
2025 Overexpression of miR-20a targeting DUSP3 inhibits OCLN ubiquitination levels and alleviates sepsis induced intestinal barrier dysfunction. In vitro cellular & developmental biology. Animal 0 40392484
2025 [Construction of etiological diagnosis model for pathogen-negative pulmonary tuberculosis using tuberculosis scores of GBP5, DUSP3, and TBP genes combined with inflammatory factors]. Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine] 0 41287336

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