Affinage

WNK3

Serine/threonine-protein kinase WNK3 · UniProt Q9BYP7

Length
1800 aa
Mass
198.4 kDa
Annotated
2026-06-11
38 papers in source corpus 26 papers cited in narrative 26 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 6/6 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

WNK3 is a serine-threonine protein kinase that functions as a coordinate regulator of cation-chloride cotransporters, setting intracellular chloride concentration and cell volume by reciprocally controlling Na-(K)-Cl cotransporters and K-Cl cotransporters: kinase-active WNK3 activates NKCC1, NKCC2 and NCC by increasing their surface expression and stimulatory-site phosphorylation while inhibiting KCC1–KCC4, and the catalytically inactive kinase produces the opposite effects, bypassing the normal tonicity requirement for transporter activation (PMID:16275911, PMID:16275913, PMID:16446421). Its activation of cotransporters operates upstream of the SPAK/OSR1 kinases through RFxV-type binding motifs, where site-specific mutations (F242A, F1337A) selectively abolish effects on individual cotransporters, and WNK3 promotes target phosphorylation by binding both SPAK and the cotransporter (PMID:18550832, PMID:22415098). WNK3 itself is regulated by extracellular tonicity/cell volume rather than by direct intracellular chloride sensing, distinguishing it from chloride-sensitive WNK4 (PMID:32579473). In neurons WNK3 controls GABAergic signaling by restraining KCC2 via inhibitory phosphorylation, so its loss hyperpolarizes the GABAA reversal potential and lowers intracellular chloride, and pathogenic catalytic-domain missense variants that impair KCC2 Thr1007 phosphorylation cause X-linked intellectual disability (PMID:34858138, PMID:36311015, PMID:35678782). The WNK3-SPAK-NKCC1 cascade also drives ischemic brain injury, as WNK3 or SPAK deletion reduces infarct volume, cerebral edema and demyelination after stroke (PMID:26069258, PMID:26861815). Beyond its core cotransporter role, WNK3 regulates ROMK1 surface expression through a kinase-independent C-terminal mechanism (PMID:16357011), activates TRPV5/TRPV6 calcium channels via the secretory pathway (PMID:18768590), modulates renal NCC abundance through ERK1/2 and NEDD4-2 (PMID:24920754, PMID:27467688), promotes cell survival through interaction with procaspase-3 (PMID:16501604), regulates neuronal Fox-1 splicing-factor localization (PMID:23027929), controls tumor PD-L1 via JNK/c-JUN signaling (PMID:36357569), and phosphorylates the circadian protein PER1 (PMID:35273702); in vivo, however, renal cotransporter regulation is largely dispensable, with WNK1 compensating for WNK3 loss in the kidney (PMID:23213404, PMID:23427142).

Mechanistic history

Synthesis pass · year-by-year structured walk · 19 steps
  1. 2005 High

    Established WNK3 as a bidirectional switch over chloride transport, answering whether a single kinase could coordinately activate Na-coupled and inhibit K-coupled chloride cotransporters.

    Evidence Kinase-active and kinase-inactive WNK3 expressed with NKCC1/NKCC2/NCC and KCC1–KCC2 in Xenopus oocytes with phosphorylation and surface-expression readouts

    PMID:16275911 PMID:16275913

    Open questions at the time
    • Performed in a heterologous oocyte system; physiological relevance in native tissue untested at this stage
    • Direct vs. indirect phosphorylation of transporters not distinguished
  2. 2006 High

    Showed WNK3 regulates KCCs through a phosphatase-dependent pathway, clarifying that WNK3 opposes the cell-volume-dependent dephosphorylation that activates K-Cl cotransport.

    Evidence Oocyte expression of KCC1–KCC4 with kinase-inactive WNK3 and PP1/PP2B inhibitors (calyculin A, cyclosporine A)

    PMID:16446421

    Open questions at the time
    • Identity of the responsible phosphatase not pinned down
    • Whether WNK3 acts directly on the phosphatase or on the transporter unresolved
  3. 2005 High

    Defined a kinase-independent function of WNK3, distinguishing its catalytic activity from a separate C-terminal mechanism that suppresses ROMK1 surface expression.

    Evidence Coexpression of ROMK1 with kinase-inactive and domain-mutant WNK3 in oocytes, with paracellular and ENaC controls

    PMID:16357011

    Open questions at the time
    • C-terminal effector mediating ROMK1 trafficking not identified
    • Physiological role in renal K+ handling untested
  4. 2006 Medium

    Demonstrated endogenous WNK3 kinase activity and a pro-survival role, expanding WNK3 function beyond ion transport into apoptosis regulation.

    Evidence IP-kinase assay, overexpression/RNAi in HeLa cells with caspase-3 readout, and co-IP with procaspase-3 and HSP70

    PMID:16501604

    Open questions at the time
    • Whether WNK3 phosphorylates procaspase-3 not established
    • Single-lab co-IP without reciprocal or structural validation
  5. 2008 High

    Established WNK3 as an upstream component of a chloride-sensitive SPAK cascade and mapped phosphorylation sites, defining the signaling architecture controlling NKCC2.

    Evidence Oocyte expression with NKCC2 threonine-to-alanine mutagenesis, SPAK-binding motif deletion, and kinase-inactive WNK3

    PMID:18550832

    Open questions at the time
    • Direct phosphorylation of SPAK by WNK3 not shown biochemically here
    • In vivo chloride-sensing mechanism unresolved
  6. 2008 High

    Localized the activating-vs-inhibiting determinant to the WNK3 amino-terminal domain and showed kinase-dependent positive regulation of TRPV5/TRPV6, broadening WNK3 targets to calcium channels.

    Evidence WNK3/WNK4 chimeras and NCC assays; oocyte TRPV5/TRPV6 Ca2+ uptake with D294A mutant, domain truncation, and colchicine

    PMID:18701621 PMID:18768590

    Open questions at the time
    • Molecular basis by which the N-terminus dictates activating vs inhibiting output not defined
    • Whether TRPV channels are direct substrates unknown
  7. 2009 Medium

    Revealed isoform-specific and SPAK-dependence differences, showing renal and brain WNK3 splice variants exert opposite effects on NCC through distinct pathways.

    Evidence Oocyte expression of renal vs brain WNK3 isoforms with kinase-dead WNK3 and kinase-dead SPAK coexpression

    PMID:19470686

    Open questions at the time
    • Mechanism by which exons 18/22 invert function not resolved
    • Tissue isoform expression patterns not validated in vivo
  8. 2011 Medium

    Tested the in vivo requirement for WNK3 in renal transporter regulation and found it largely dispensable, prompting a compensation model.

    Evidence WNK3 knockout mice with immunoblotting of pSPAK/OSR1/NKCC2/NCC, urine electrolytes, and blood pressure under salt diets

    PMID:23213404

    Open questions at the time
    • Compensatory mechanism only inferred from WNK1 upregulation
    • Possible developmental compensation not excluded
  9. 2011 Medium

    Linked WNK3-NKCC1 signaling to a disease-relevant cellular behavior, showing WNK3 supports volume regulation and invasion in glioma.

    Evidence Co-IP of WNK3 with NKCC1 plus shRNA knockdown with bumetanide-sensitive volume and Transwell invasion assays

    PMID:21813709

    Open questions at the time
    • Whether invasion effect is solely NKCC1-dependent unclear
    • Single-lab functional data
  10. 2012 High

    Mapped the RFxV SPAK-binding motifs that govern target selectivity and identified a non-canonical role in mRNA splicing, defining how WNK3 selects among cotransporters and uncovering Fox-1 regulation.

    Evidence Systematic F242A/F1337A/F873A mutagenesis with cotransporter assays; co-IP, in vitro phosphorylation, and splicing reporters for Fox-1

    PMID:22415098 PMID:23027929

    Open questions at the time
    • How the same motifs confer different specificity for each cotransporter not fully explained
    • Fox-1 splicing role tested in heterologous reporter context only
  11. 2013 High

    Placed WNK3 downstream of angiotensin II/AT1 signaling in vasculature and identified an inhibitory kinase regulator, situating WNK3 within hormonal blood-pressure control and neuronal apoptosis.

    Evidence WNK3 KO mice with salt/AngII manipulation and valsartan; LINGO-1 co-IP, Nogo66 stimulation, and kinase-activity assays in neurons

    PMID:23482566 PMID:24019400

    Open questions at the time
    • Direct vascular substrate of the WNK3-SPAK-NKCC1 cascade not defined
    • Mechanism by which LINGO-1 binding inhibits kinase activity unresolved
  12. 2013 Medium

    Confirmed WNK1/SPAK axis compensation for renal WNK3 loss, reconciling robust oocyte effects with a minor in vivo kidney phenotype.

    Evidence WNK3 KO mice with immunoblotting of WNK1/WNK4 and phospho-transporters plus hydrochlorothiazide response under salt diets

    PMID:23427142

    Open questions at the time
    • Mechanism of WNK1 upregulation upon WNK3 loss not established
    • Extent of functional redundancy across nephron segments unclear
  13. 2014 Medium

    Identified NEDD4-2 as an N-terminal partner and showed WNK3 reverses NEDD4-2-mediated NCC inhibition through a non-catalytic, SPAK-independent route, distinguishing this from its canonical cascade.

    Evidence GST pulldown/LC-MS/MS and co-IP in HEK293; ubiquitylation/phosphorylation assays and oocyte assays with WNK3-F242A

    PMID:24920754

    Open questions at the time
    • Molecular mechanism by which WNK3 antagonizes NEDD4-2 not defined
    • Single-lab interaction without structural mapping
  14. 2015 High

    Established WNK3-SPAK/OSR1-NKCC1 as the operative cascade driving ischemic brain edema and injury, providing a therapeutic rationale.

    Evidence WNK3 KO mice with MCAO, infarct/edema/demyelination readouts, cascade phospho-immunoblotting, and OGD with SPAK/OSR1 siRNA in neurons/oligodendrocytes

    PMID:26069258

    Open questions at the time
    • Upstream trigger activating WNK3 during ischemia not identified
    • Cell-type-specific contributions not fully dissected
  15. 2016 High

    Uncovered a transcriptional/translational arm of WNK3 NCC regulation via ERK1/2 and confirmed the WNK3-SPAK complex as the stroke effector unit using complementary genetics.

    Evidence Overexpression/knockdown in DCT cells with CHX pulse-chase and ERK1/2 knockdown; WNK3 KO, SPAK het and SPAK KO mice with MCAO and DTI imaging

    PMID:26861815 PMID:27467688

    Open questions at the time
    • How WNK3 lowers ERK1/2 phosphorylation mechanistically unknown
    • Whether ERK1/2 arm operates in vivo not tested
  16. 2020 Medium

    Redefined WNK3 regulation as cell-volume/tonicity-responsive rather than intracellular-chloride-sensitive, separating it mechanistically from WNK4.

    Evidence Oocyte NCC assays with WNK3 chloride-binding-site mutagenesis and WNK3/WNK4 chimeras under controlled tonicity and Cl-

    PMID:32579473

    Open questions at the time
    • The volume-sensing element within the C-terminus not identified
    • Physiological signal coupling volume to WNK3 activity unknown
  17. 2021 Medium

    Demonstrated WNK3 controls GABAergic chloride homeostasis in mature neurons via KCC2 phosphorylation, defining its role in synaptic inhibition development.

    Evidence shRNA knockdown in hippocampal neurons with perforated patch-clamp EGABA, KCC2 phospho-immunoblotting, and KCC2 inhibitor VU024055 rescue

    PMID:34858138

    Open questions at the time
    • Why the effect is age-dependent (mature vs immature) not explained
    • Direct WNK3 phosphorylation of KCC2 not shown biochemically here
  18. 2022 Medium

    Extended WNK3-KCC2 regulation to intrinsic excitability and resting K+ conductance in cortical neurons and tied catalytic-domain variants to X-linked intellectual disability, establishing a disease mechanism.

    Evidence WNK3 KO mice with patch-clamp, active-WNK3 reintroduction, CLP290 rescue, and behavioral testing; ID-family sequencing with KCC2 T1007 phosphorylation assays of variants

    PMID:35678782 PMID:36311015

    Open questions at the time
    • Link between altered Cl-/excitability and cognitive phenotype not directly demonstrated
    • Variant functional assays in patient-derived neurons not performed
  19. 2022 Low

    Identified additional kinase-dependent roles in tumor immune evasion and circadian regulation, broadening WNK3 substrates beyond ion transport.

    Evidence shRNA screen, kinase-dead mutant, WNK463 inhibitor and MC38 model with c-JUN analysis for PD-L1; co-IP and in vitro PER1 phosphorylation with SCN shRNA knockdown and sleep analysis

    PMID:35273702 PMID:36357569

    Open questions at the time
    • PER1/circadian role rests on a single low-confidence study without reconstitution
    • How WNK3 engages the JNK/c-JUN pathway to regulate PD-L1 transcription not mechanistically defined

Open questions

Synthesis pass · forward-looking unresolved questions
  • How extracellular tonicity is transduced into WNK3 catalytic activation, and how a single kinase achieves opposite, target-specific outputs across cotransporters, channels, and splicing/transcriptional substrates, remains unresolved.
  • No structural model of the volume-sensing C-terminus
  • Mechanism converting one kinase into bidirectional transporter control unknown
  • Direct vs indirect phosphorylation for several proposed substrates undetermined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 6 GO:0016740 transferase activity 4 GO:0098772 molecular function regulator activity 3
Localization
GO:0005634 nucleus 1 GO:0005829 cytosol 1
Pathway
R-HSA-382551 Transport of small molecules 5 R-HSA-112316 Neuronal System 3 R-HSA-162582 Signal Transduction 3 R-HSA-5357801 Programmed Cell Death 2
Partners
KCC2LINGO-1NCCNEDD4-2NKCC1OSR1PER1SPAK
Complex memberships
WNK3-SPAK kinase complex

Evidence

Reading pass · 26 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2005 Kinase-active WNK3 increases Cl- influx via NKCC1 and inhibits Cl- exit through KCC1 and KCC2; kinase-inactive WNK3 has opposite effects. WNK3 effects are imparted via altered phosphorylation and surface expression of its downstream targets and bypass the normal requirement of altered tonicity for activation of these transporters. Heterologous expression in Xenopus laevis oocytes; kinase-inactive mutant studies; phosphorylation and surface expression assays Proceedings of the National Academy of Sciences of the United States of America High 16275911
2005 Kinase-active WNK3 potently activates both NKCC2 and NCC-mediated transport; kinase-inactive WNK3 potently inhibits both. WNK3 regulates these transporters by altering their expression at the plasma membrane and increases phosphorylation of NKCC2 at Thr-184 and Thr-189. Coexpression studies in Xenopus oocytes; kinase-inactive mutant; plasma membrane expression assays; phosphorylation assays Proceedings of the National Academy of Sciences of the United States of America High 16275913
2006 WNK3 completely prevents cell swelling-induced activation of KCC1-KCC4 in Xenopus oocytes. Catalytically inactive WNK3 abolishes cell shrinkage-induced inhibition of KCC1-KCC4, resulting in >100-fold stimulation of K-Cl cotransport. This activation is abolished by calyculin A and cyclosporine A (protein phosphatase 1 and 2B inhibitors), indicating WNK3 acts via a phosphatase-dependent pathway to regulate KCCs. Heterologous expression in Xenopus oocytes; kinase-inactive mutant; pharmacological phosphatase inhibitors (calyculin A, cyclosporine A) Proceedings of the National Academy of Sciences of the United States of America High 16446421
2008 Intracellular chloride depletion activates NKCC2 by promoting phosphorylation of three conserved threonines (T96, T101, T111). This chloride-sensitive activation requires WNK3 upstream of SPAK; elimination of WNK3's SPAK-binding motif prevents NKCC2 activation, as does catalytically inactive WNK3. Heterologous expression in Xenopus laevis oocytes; KCC2 co-expression and hypotonic stress for Cl- depletion; threonine-to-alanine mutagenesis; SPAK-binding motif deletion; kinase-inactive mutant Proceedings of the National Academy of Sciences of the United States of America High 18550832
2005 WNK3 inhibits the renal K+ channel ROMK1 >5.5-fold by reducing its plasmalemmal surface expression. This inhibition is independent of WNK3's catalytic activity and is mediated by WNK3's carboxyl terminus. A kinase-inactivating point mutation enhances ROMK1 inhibition >2.5-fold. WNK3 had no effect on paracellular Cl- flux or ENaC-mediated currents. Coexpression in Xenopus laevis oocytes; kinase-inactive and domain mutants; MDCK II paracellular flux assay; surface expression analysis The Journal of physiology High 16357011
2006 Endogenous WNK3 is an active protein kinase when immunoprecipitated from cells. WNK3 overexpression increases HeLa cell survival by delaying apoptosis onset; WNK3 knockdown by RNAi accelerates apoptotic response and promotes caspase-3 activation. The mechanism involves interaction of WNK3 with procaspase-3 and heat-shock protein 70. Immunoprecipitation kinase assay; overexpression; RNAi knockdown; caspase-3 activation assays; co-immunoprecipitation with procaspase-3 and HSP70 Oncogene Medium 16501604
2008 WNK3 positively regulates TRPV5 and TRPV6 epithelial calcium channels in a kinase-dependent manner, increasing TRPV5-mediated Ca2+ uptake by ~83%. The kinase domain alone is sufficient; D294A kinase-inactive mutation abolishes the effect. WNK3 increases the complexly glycosylated, plasma membrane-expressed form of TRPV5 via enhanced delivery through the secretory pathway, blocked by microtubule inhibitor colchicine. Coexpression in Xenopus laevis oocytes; kinase-inactive D294A mutant; kinase domain truncation; Ca2+ uptake assay; voltage-clamp; plasma membrane expression and exocytosis assays; colchicine treatment American journal of physiology. Renal physiology High 18768590
2008 The amino-terminal domain of WNK3 (not the kinase or carboxyl domain) determines whether a WNK kinase activates or inhibits NCC. Chimeras swapping amino- or carboxyl-terminal domains between WNK3 and WNK4 show that the amino terminus governs activating vs. inhibiting properties toward NCC. WNK3/WNK4 chimera construction and coexpression in Xenopus oocytes; NCC transport assays American journal of physiology. Renal physiology Medium 18701621
2009 Renal and brain splice isoforms of WNK3 (differing by exons 18 and 22) have opposite effects on NCC: the renal isoform increases NCC expression/activity while the brain isoform decreases it. Kinase-inactivating mutation in renal WNK3 reverses its effect. Brain WNK3 acts in tandem with SPAK, whereas renal WNK3 upregulates NCC through a SPAK-independent pathway. Expression in Xenopus oocytes; kinase-dead mutant; kinase-dead SPAK coexpression; NCC surface expression and transport assays Journal of the American Society of Nephrology : JASN Medium 19470686
2011 WNK3 knockout mice show no significant decrease in phosphorylation or expression of OSR1, SPAK, NKCC2, or NCC under normal or low-salt diets, indicating WNK3 has only a minor role in regulating these transporters in vivo in the kidney. WNK1 expression was slightly elevated in knockout mice on low-salt diet, suggesting compensatory upregulation. WNK3 knockout mouse generation; immunoblotting for pOSR1, pSPAK, pNKCC2, pNCC; urine electrolyte measurements; blood pressure measurement Biology open Medium 23213404
2011 In glioma cells, WNK3 co-localizes and co-immunoprecipitates with NKCC1 upon cell volume changes. Stable knockdown of WNK3 completely abolishes bumetanide-sensitive NKCC1 activity and reduces invasive capacity across Transwell barriers. Co-immunoprecipitation; shRNA knockdown; bumetanide-sensitive cell volume regulation assay; Transwell invasion assay American journal of physiology. Cell physiology Medium 21813709
2012 WNK3 binds Fox-1 (neuronal mRNA splicing factor) and inhibits its splicing activity in a kinase activity-dependent manner. WNK3 phosphorylates Fox-1 without changing its RNA binding capacity; instead, WNK3 increases the cytoplasmic localization of Fox-1, thereby suppressing Fox-1-dependent splicing. Co-immunoprecipitation; in vitro phosphorylation assay; subcellular localization studies; splicing reporter assays; kinase-inactive mutant comparisons Proceedings of the National Academy of Sciences of the United States of America Medium 23027929
2012 WNK3's effects on NCC and other CCCs require interaction with SPAK via specific RFxV binding motifs. The F242A mutation in the first SPAK-binding site of WNK3 abolishes effects on all tested cotransporters; F1337A eliminates effect on NKCC2 but preserves effects on NCC, NKCC1, and KCC4; F873A has no consequence. WNK3 can promote NCC phosphorylation at T58 even without the NCC SPAK-binding site, suggesting WNK3 binds both SPAK and NCC to promote phosphorylation. Site-directed mutagenesis of SPAK binding motifs; expression in Xenopus laevis oocytes; transport assays; phosphorylation assays Cellular physiology and biochemistry High 22415098
2013 In mouse aorta, the WNK3-SPAK-NKCC1 phosphorylation cascade is regulated by dietary salt intake through angiotensin II acting via the AT1 receptor. In WNK3 knockout mice, low-salt diet and angiotensin II infusion fail to increase SPAK/NKCC1 phosphorylation, demonstrating WNK3 dependence. Angiotensin II-induced blood pressure increases are diminished in WNK3 knockout mice. WNK3 knockout mice; dietary salt manipulation; angiotensin II infusion; AT1 receptor antagonist (valsartan); immunoblotting for pSPAK, pNKCC1; blood pressure measurement Hypertension High 24019400
2013 LINGO-1 co-localizes and co-precipitates with WNK3 in cultured cortical neurons and brain tissue. Nogo66 stimulation enhances this co-association and reduces WNK3 kinase activity. LINGO-1 binding to WNK3's intracellular domain reduces WNK3 kinase activity, thereby promoting neuronal apoptosis. Co-immunoprecipitation; co-localization immunofluorescence; kinase activity assay; shRNA knockdown; Nogo66 stimulation; LINGO-1 intracellular domain construct The Journal of biological chemistry Medium 23482566
2013 In WNK3-deficient mice, WNK1 is markedly upregulated while WNK4 expression remains similar, indicating that WNK1/SPAK axis compensates for WNK3 loss in the kidney. Under low-salt diet, pSPAK/OSR1, pNKCC2, and pNCC levels in WNK3-/- exceed those in wild-type, confirming compensatory activation of WNK3-independent pathways. WNK3 knockout mice; immunoblotting for WNK1, WNK4, pSPAK/OSR1, pNKCC2, pNCC; hydrochlorothiazide diuretic response; dietary salt manipulation American journal of physiology. Renal physiology Medium 23427142
2014 NEDD4-2 was identified as a WNK3-interacting partner via GST pulldown of WNK3's N-terminal domain followed by LC-MS/MS, validated by co-immunoprecipitation in HEK293 cells. WNK3 interaction with NEDD4-2 does not involve the PY-like motif. WNK3 does not ubiquitylate NEDD4-2 or phosphorylate NEDD4-2 on S222/S328, and does not regulate NCC-NEDD4-2 interaction. WNK3 can overcome SGK1-resistant NEDD4-2-mediated inhibition of NCC via a SPAK-independent mechanism (F242A mutation blocks NCC activation but not NEDD4-2 inhibitory reversal). GST pulldown with LC-MS/MS; co-immunoprecipitation in HEK293 cells; ubiquitylation assay; phosphorylation assay; Xenopus oocyte expression with SPAK-binding mutant WNK3-F242A American journal of physiology. Renal physiology Medium 24920754
2015 In ischemic stroke (transient MCAO), WNK3 knockout mice exhibit significantly decreased infarct volume and axonal demyelination, reduced cerebral edema, and accelerated neurological recovery. The neuroprotective phenotype is associated with decreased hyperphosphorylation of SPAK/OSR1 catalytic T-loop and of NKCC1 stimulatory sites Thr203/Thr207/Thr212, and decreased cell surface expression of NKCC1. siRNA knockdown of SPAK/OSR1 also increased tolerance of neurons and oligodendrocytes to in vitro ischemia, confirming WNK3-SPAK/OSR1-NKCC1 as the operative cascade. WNK3 knockout mice; transient MCAO; infarct volume, edema, demyelination assessment; immunoblotting and immunostaining for pSPAK/OSR1, pNKCC1, NKCC1 surface expression; oxygen-glucose deprivation/reoxygenation in neurons and oligodendrocytes; siRNA knockdown Stroke High 26069258
2016 WNK3 enhances NCC protein expression by increasing NCC protein synthesis (not via lysosomal degradation pathway). WNK3 reduces ERK1/2 phosphorylation, and knockdown of ERK1/2 expression reverses WNK3-mediated increase in NCC, indicating WNK3 acts through an ERK1/2 signaling pathway to upregulate NCC. Overexpression and siRNA knockdown in Cos-7 and mouse distal convoluted tubule cells; cycloheximide pulse-chase assay; bafilomycin A1 treatment; ERK1/2 knockdown; Western blot Nephron Medium 27467688
2016 Deletion of the WNK3-SPAK kinase complex in mice (WNK3 KO and SPAK heterozygous or KO) each produce >50% reduction in infarct size and cerebral edema, significantly less demyelination, and improved neurological outcomes after MCAO stroke, establishing the WNK3-SPAK complex as the operative signaling unit mediating ischemic brain injury. WNK3 KO, SPAK heterozygous and SPAK KO mice; transient MCAO; MRI diffusion tensor imaging; immunofluorescence; neurological deficit scoring up to 14 days Journal of cerebral blood flow and metabolism High 26861815
2020 WNK3 is not sensitive to intracellular chloride concentration but is regulated by changes in extracellular tonicity (cell volume changes). Elimination of the chloride-binding site in WNK3 does not affect its activity toward NCC. Chimeric WNK3/WNK4 analysis suggests sequences within the carboxy-terminal end modulate chloride affinity. In contrast, WNK4 is chloride-sensitive but not regulated by cell volume. Xenopus oocyte expression system; chloride-binding site mutagenesis; WNK3/WNK4 chimeric protein constructs; tonicity and Cl- concentration manipulation; NCC transport assays American journal of physiology. Cell physiology Medium 32579473
2021 WNK3 knockdown in hippocampal neurons hyperpolarizes the GABAA reversal potential (EGABA) in mature (DIV13-15) but not immature (DIV6-8) neurons. This effect is mediated through reduced phosphorylation of KCC2 at the membrane (increased KCC2 activity), reversed by KCC2 inhibitor VU024055. WNK3 knockdown also causes morphological changes and increased intrinsic excitability in mature neurons. shRNA knockdown in embryonic hippocampal neurons; gramicidin-perforated patch-clamp; immunoblotting for KCC2 phosphorylation; KCC2 inhibitor (VU024055); membrane surface expression analysis Frontiers in molecular neuroscience Medium 34858138
2022 WNK3 knockout mice show elevated intracellular Cl- in layer V prefrontal cortex pyramidal neurons (EGABA depolarized by 6 mV, corresponding to ~4 mM higher [Cl-]i). KO neurons exhibit increased inwardly rectifying K+ (Kir channel) conductance, hyperpolarized resting membrane potential, and increased firing threshold. Introducing active WNK3 into KO neurons reverses these changes. The KCC2 activator CLP290 reverses enhanced IRK conductance in KO neurons, linking WNK3 and KCC2 in resting K+ conductance regulation. WNK3 KO mice show behavioral deficits in prepulse inhibition. WNK3 KO mice; gramicidin-perforated patch-clamp in acute slices; intracellular active WNK3 injection; CLP290 (KCC2 activator); immunoblotting for pSPAK, pOSR1, pNKCC1, pKCC2; mEPSC/mIPSC recording; prepulse inhibition behavioral test Frontiers in molecular neuroscience Medium 36311015
2022 WNK3 pathogenic missense variants (p.Pro204Arg, p.Leu300Ser, p.Glu607Val) localize to the catalytic domain and impede the inhibitory phosphorylation of neuronal KCC2 at threonine 1007, a site critically regulated during development of synaptic inhibition, causing X-linked intellectual disability. Exome/genome sequencing of intellectual disability families; functional assays of KCC2 T1007 phosphorylation with disease-associated WNK3 variants Genetics in medicine Medium 35678782
2022 WNK3 is a positive regulator of PD-L1 expression in tumor cells via a kinase-dependent mechanism. Kinase-dead WNK3 mutant fails to elevate PD-L1 levels. WNK3 regulates PD-L1 transcription through the JNK/c-JUN pathway. Pooled shRNA screening; kinase-dead mutant; co-culture of cancer and immune cells; cytokine and cytolytic enzyme secretion assays; WNK463 pan-WNK inhibitor; MC38 syngeneic mouse tumor model; c-JUN pathway analysis Experimental & molecular medicine Medium 36357569
2022 WNK3 co-localizes and interacts with the circadian clock protein PER1 in the suprachiasmatic nucleus. WNK3 phosphorylates PER1 to promote its degradation. WNK3 knockdown in rat SCN alters sleep patterns. Co-localization and co-immunoprecipitation in SCN; in vitro kinase assay (PER1 phosphorylation); shRNA knockdown in vivo; sleep pattern analysis American journal of translational research Low 35273702

Source papers

Stage 0 corpus · 38 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2008 Regulation of NKCC2 by a chloride-sensing mechanism involving the WNK3 and SPAK kinases. Proceedings of the National Academy of Sciences of the United States of America 195 18550832
2005 WNK3 modulates transport of Cl- in and out of cells: implications for control of cell volume and neuronal excitability. Proceedings of the National Academy of Sciences of the United States of America 175 16275911
2005 WNK3 kinase is a positive regulator of NKCC2 and NCC, renal cation-Cl- cotransporters required for normal blood pressure homeostasis. Proceedings of the National Academy of Sciences of the United States of America 152 16275913
2006 WNK3 bypasses the tonicity requirement for K-Cl cotransporter activation via a phosphatase-dependent pathway. Proceedings of the National Academy of Sciences of the United States of America 104 16446421
2015 Inhibition of WNK3 Kinase Signaling Reduces Brain Damage and Accelerates Neurological Recovery After Stroke. Stroke 86 26069258
2011 With-No-Lysine Kinase 3 (WNK3) stimulates glioma invasion by regulating cell volume. American journal of physiology. Cell physiology 56 21813709
2005 WNK3, a kinase related to genes mutated in hereditary hypertension with hyperkalaemia, regulates the K+ channel ROMK1 (Kir1.1). The Journal of physiology 51 16357011
2013 Dietary salt intake regulates WNK3-SPAK-NKCC1 phosphorylation cascade in mouse aorta through angiotensin II. Hypertension (Dallas, Tex. : 1979) 50 24019400
2004 Cloning, genomic organization, alternative splicing and expression analysis of the human gene WNK3 (PRKWNK3). Gene 50 15194194
2011 A minor role of WNK3 in regulating phosphorylation of renal NKCC2 and NCC co-transporters in vivo. Biology open 45 23213404
2009 Renal and brain isoforms of WNK3 have opposite effects on NCCT expression. Journal of the American Society of Nephrology : JASN 45 19470686
2006 Protein kinase WNK3 increases cell survival in a caspase-3-dependent pathway. Oncogene 40 16501604
2016 Deletion of the WNK3-SPAK kinase complex in mice improves radiographic and clinical outcomes in malignant cerebral edema after ischemic stroke. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism 34 26861815
2011 WNK3 is a putative chloride-sensing kinase. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 34 22179001
2008 WNK3 and WNK4 amino-terminal domain defines their effect on the renal Na+-Cl- cotransporter. American journal of physiology. Renal physiology 33 18701621
2008 WNK3 positively regulates epithelial calcium channels TRPV5 and TRPV6 via a kinase-dependent pathway. American journal of physiology. Renal physiology 33 18768590
2013 Loss of WNK3 is compensated for by the WNK1/SPAK axis in the kidney of the mouse. American journal of physiology. Renal physiology 30 23427142
2012 WNK3-SPAK interaction is required for the modulation of NCC and other members of the SLC12 family. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 30 22415098
2020 Activated WNK3 induced by intracerebral hemorrhage deteriorates brain injury maybe via WNK3/SPAK/NKCC1 pathway. Experimental neurology 28 32589890
2011 Similar effects of all WNK3 variants on SLC12 cotransporters. American journal of physiology. Cell physiology 24 21613606
2012 Influence of WNK3 on intracellular chloride concentration and volume regulation in HEK293 cells. Pflugers Archiv : European journal of physiology 23 22864523
2020 WNK3 and WNK4 exhibit opposite sensitivity with respect to cell volume and intracellular chloride concentration. American journal of physiology. Cell physiology 22 32579473
2014 WNK3 abrogates the NEDD4-2-mediated inhibition of the renal Na+-Cl- cotransporter. American journal of physiology. Renal physiology 22 24920754
2021 Silencing of the lncRNA H19 enhances sensitivity to X-ray and carbon-ions through the miR-130a-3p /WNK3 signaling axis in NSCLC cells. Cancer cell international 21 34863180
2013 LINGO-1 receptor promotes neuronal apoptosis by inhibiting WNK3 kinase activity. The Journal of biological chemistry 21 23482566
2012 Protein kinase WNK3 regulates the neuronal splicing factor Fox-1. Proceedings of the National Academy of Sciences of the United States of America 16 23027929
2022 WNK3 inhibition elicits antitumor immunity by suppressing PD-L1 expression on tumor cells and activating T-cell function. Experimental & molecular medicine 8 36357569
2018 Increased expression of WNK3 in dispersed granule cells in hippocampal sclerosis of mesial temporal lobe epilepsy patients. Epilepsy research 8 30253317
2022 Rare pathogenic variants in WNK3 cause X-linked intellectual disability. Genetics in medicine : official journal of the American College of Medical Genetics 7 35678782
2021 WNK3 Promotes Neuronal Survival after Traumatic Brain Injury in Rats. Neuroscience 4 34626749
2022 WNK3 kinase maintains neuronal excitability by reducing inwardly rectifying K+ conductance in layer V pyramidal neurons of mouse medial prefrontal cortex. Frontiers in molecular neuroscience 3 36311015
2021 WNK3 Maintains the GABAergic Inhibitory Tone, Synaptic Excitation and Neuronal Excitability via Regulation of KCC2 Cotransporter in Mature Neurons. Frontiers in molecular neuroscience 3 34858138
2016 WNK3 Kinase Enhances the Sodium Chloride Cotransporter Expression via an ERK 1/2 Signaling Pathway. Nephron 3 27467688
2022 Kinome-Wide Profiling Identifies Human WNK3 as a Target of Cajanin Stilbene Acid from Cajanus cajan (L.) Millsp. International journal of molecular sciences 2 35163434
2022 WNK3-PER1 interactions regulate the circadian rhythm in the suprachiasmatic nucleus in rats. American journal of translational research 2 35273702
2021 WNK3 promotes the invasiveness of glioma cell lines under hypoxia by inducing the epithelial-to-mesenchymal transition. Translational neuroscience 2 34513083
2025 ZFHX3 silencing alleviates ischemic stroke by suppressing pericyte contraction and calcium influx through inhibiting WNK3 expression. Experimental neurology 1 40602592
2025 Increased expression of WNK3 during the perinatal period in newborn rats with hypoxic-ischemic encephalopathy. Brain injury 0 40993905

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