Affinage

NFAT5

Nuclear factor of activated T-cells 5 · UniProt O94916

Length
1531 aa
Mass
165.8 kDa
Annotated
2026-04-29
100 papers in source corpus 43 papers cited in narrative 42 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

NFAT5 (TonEBP/OREBP) is a Rel-family transcription factor that functions as a master osmostress-responsive transactivator, an NF-κB/AP-1 transcriptional cofactor in immune and inflammatory signaling, and a guardian of genome integrity through R-loop resolution. Structurally, NFAT5 homodimerizes via its Rel homology domain and completely encircles asymmetric TonE DNA elements, conferring kinetically stable DNA binding; its activity is governed by regulated nuclear-cytoplasmic shuttling through a CRM1-dependent NES, phosphorylation of Y143 by c-Abl (opposed by SHP-1 phosphatase) to recruit PLC-γ1, phosphorylation of T135 by CDK5 for rapid nuclear import, and mRNA stabilization via its 5′-UTR under hypertonicity (PMID:11780147, PMID:16782704, PMID:20585028, PMID:20351292, PMID:21209322, PMID:15900024). Beyond classical osmoadaptive targets (AQP1, AQP2, UT-A, aldose reductase), NFAT5 acts as a DNA-binding-independent cofactor that recruits p300 to NF-κB enhanceosomes in macrophages and microglia, represses IFNβ by competing with IRF3 at the IFNB1 enhanceosome, promotes CD8⁺ T cell exhaustion via TOX/PD-1 induction in the hyperosmolar tumor microenvironment, and supports thymocyte survival at the β-selection checkpoint (PMID:27118681, PMID:31816635, PMID:37709986, PMID:24043824). NFAT5 also recognizes R-loop structures and recruits METTL3 for m⁶A-mediated R-loop resolution, a process dependent on PARP1-mediated PARylation, linking it to DNA damage repair (PMID:33313823, PMID:34049076).

Mechanistic history

Synthesis pass · year-by-year structured walk · 16 steps
  1. 1999 High

    Identification of NFAT5 as a divergent, constitutively nuclear Rel-family member established that it represents a distinct branch of the NFAT/Rel superfamily that does not rely on calcineurin-regulated nuclear import or cooperative Fos/Jun binding.

    Evidence DNA binding assays, nuclear fractionation, and sequence analysis in transfected cells

    PMID:10377394

    Open questions at the time
    • Calcineurin-independent mechanism of nuclear localization not yet defined
    • Endogenous genomic targets unknown
  2. 2000 High

    Demonstration that NFAT5 directly transactivates osmoprotective genes (UT-A urea transporters) through TonE elements established NFAT5 as the effector transcription factor of the tonicity-responsive pathway, while calcineurin-dependent induction of NFAT5 expression during T cell activation revealed a second, osmostress-independent regulatory axis.

    Evidence TonE promoter mutagenesis, dominant-negative TonEBP, pharmacological calcineurin inhibition in T cells

    PMID:10995747 PMID:11046013

    Open questions at the time
    • Full repertoire of TonE-dependent target genes not mapped
    • Mechanism linking calcineurin to NFAT5 gene induction unclear
  3. 2002 High

    Crystal structure of the TonEBP-DNA complex revealed that NFAT5 homodimers completely encircle asymmetric DNA, explaining the kinetic stability of its DNA engagement and confirming an NF-κB-like fold; parallel work showed that posttranslational phosphorylation controls transactivation domain (TAD) activity in proportion to extracellular tonicity.

    Evidence X-ray crystallography with biochemical encirclement assays; GAL4-TAD reporter assays with kinase inhibitors

    PMID:11780147 PMID:11792870

    Open questions at the time
    • Identity of the kinases phosphorylating the TAD not yet determined
    • Structural basis of TAD regulation unresolved
  4. 2005 High

    Discovery that high NaCl stabilizes NFAT5 mRNA via its 5′-UTR and that mitochondrial ROS (complexes I/III) selectively enhance TAD activity revealed two post-transcriptional/signaling inputs that amplify the osmotic response independently of nuclear import.

    Evidence Actinomycin D mRNA decay assay with UTR-swapped reporters; ROS scavengers and mitochondrial complex inhibitors with reporter assays

    PMID:15769933 PMID:15900024

    Open questions at the time
    • RNA-binding factor mediating 5′-UTR stabilization not identified
    • Direct ROS target in the TAD pathway unknown
  5. 2006 High

    Mapping of a CRM1-dependent NES and an auxiliary export domain in NFAT5 defined the nuclear export arm of its shuttling, while proteomic identification of nuclear partners (Hsp90, PARP-1, DNA-PK, RNA helicases) expanded the functional network beyond simple DNA binding.

    Evidence NES mutagenesis with GFP imaging and leptomycin B; nuclear IP-MS with functional reporter validation

    PMID:16782704 PMID:17148781

    Open questions at the time
    • Nuclear import signal and its regulation not yet mapped
    • Functional significance of PARP-1 and DNA-PK interactions not clarified
  6. 2010 High

    A coherent phosphoswitch was established: c-Abl phosphorylates Y143 to promote nuclear localization and TAD activity, SHP-1 opposes this by dephosphorylating Y143, and phospho-Y143 recruits PLC-γ1 as an obligate cofactor — providing a unified mechanism linking osmosensing kinase/phosphatase balance to NFAT5 activation.

    Evidence In vitro kinase assays, genome-wide phosphatase siRNA screen, co-IP with Y143A mutant, PLC-γ1-null cell reconstitution

    PMID:20080774 PMID:20351292 PMID:20585028

    Open questions at the time
    • How high NaCl activates c-Abl is unknown
    • Whether PLC-γ1 catalytic activity or scaffolding function is required is unresolved
  7. 2011 High

    CDK5 was identified as a second kinase that phosphorylates T135 for rapid early nuclear import (without affecting TAD), and Rac1/OSM were shown to act upstream of PLC-γ1 (not p38) to support NFAT5 activity, ordering the signaling cascade from membrane GTPase to transcription factor.

    Evidence MS-based phosphosite identification, CDK5 inhibitor/siRNA, Rac1 epistasis in PLC-γ1-null MEFs with reconstitution

    PMID:21209322 PMID:21712438

    Open questions at the time
    • Relationship between CDK5-T135 and c-Abl-Y143 phosphorylation events not established
    • Full complement of upstream Rac1 activators in osmostress unknown
  8. 2010 High

    NFAT5 was shown to interact with NF-κB p65 in a tonicity- and Akt-dependent manner, binding κB elements and enhancing NF-κB-driven transcription — establishing NFAT5 as a transcriptional cofactor for inflammatory signaling beyond its classical osmoprotective role.

    Evidence Reciprocal co-IP, ChIP at κB sites, Akt inhibitor epistasis in renal epithelial cells and macrophages

    PMID:20685965

    Open questions at the time
    • Direct vs. indirect DNA binding at κB sites not fully resolved
    • Whether Akt directly phosphorylates NFAT5 is unknown
  9. 2012 High

    TAZ was identified as a phosphorylation-dependent negative regulator: c-Abl phosphorylates TAZ-Y316, enabling TAZ to bind NFAT5 and suppress its DNA binding — revealing a feedback brake on NFAT5 activation.

    Evidence Co-IP, Y316F mutagenesis, TAZ KO with WT vs. mutant rescue, EMSA

    PMID:23045390

    Open questions at the time
    • Temporal dynamics of TAZ inhibition relative to c-Abl-Y143 activation unclear
    • Whether TAZ competes with PLC-γ1 for NFAT5 binding is unknown
  10. 2013 High

    Conditional knockout revealed that NFAT5 promotes thymocyte survival at the β-selection checkpoint by upregulating A1/Bcl2 and suppressing p53/Noxa, an osmostress-independent function regulated by IKKβ — extending NFAT5 biology into adaptive immune development.

    Evidence Conditional NFAT5 KO mice, IKKβ genetic manipulation, apoptosis and gene expression analysis

    PMID:24043824

    Open questions at the time
    • Whether NFAT5 acts through TonE or NF-κB sites in thymocyte target genes not determined
    • Mechanism by which IKKβ regulates NFAT5 expression not defined
  11. 2016 High

    NFAT5 was shown to function as a DNA-binding-independent NF-κB cofactor by recruiting p300 to the LPS-induced enhanceosome in macrophages, with myeloid-specific deletion attenuating sepsis — establishing a non-classical mechanism of transcriptional coactivation.

    Evidence ChIP showing p300 recruitment loss upon TonEBP deletion, myeloid-specific KO mice in sepsis model

    PMID:27118681

    Open questions at the time
    • How NFAT5 is recruited to enhanceosome without direct DNA binding not clarified
    • Whether other coactivators besides p300 are involved is unknown
  12. 2017 High

    NFAT5 was found to drive macrophage survival in rheumatoid arthritis by inducing autocrine CCL2, with haploinsufficiency reducing joint destruction — directly linking NFAT5 to inflammatory disease pathogenesis.

    Evidence NFAT5-deficient macrophages, apoptosis assays, CCL2 rescue in Nfat5+/- mice

    PMID:28192374

    Open questions at the time
    • Whether NFAT5 binds the CCL2 promoter directly via TonE or NF-κB cofactor mechanism not resolved
    • Contribution of other NFAT5 target genes to macrophage survival unknown
  13. 2019 High

    Two studies revealed NFAT5's dual role as an epigenetic repressor: it recruits DNMT1 to methylate and silence the β3-adrenoreceptor promoter in adipocytes (opposing thermogenesis), and it blocks Nrf2 access to the HO-1 promoter (promoting M1 macrophage polarization) — establishing gene-repressive functions beyond classical transactivation.

    Evidence ChIP, bisulfite sequencing, adipocyte-specific KO (high-fat diet resistance); Nrf2 competition ChIP in macrophages

    PMID:31057560 PMID:31387996

    Open questions at the time
    • Generality of DNMT1 recruitment mechanism to other NFAT5-repressed loci unknown
    • Whether NFAT5 displaces Nrf2 by direct competition or chromatin remodeling not distinguished
  14. 2020 High

    NFAT5 was shown to repress IFNβ by competing with IRF3 at the IFNB1 enhanceosome, while simultaneously serving as a cofactor for both NF-κB and AP-1 in microglia — demonstrating context-dependent activating and repressive roles at distinct promoters within the same cell type.

    Evidence NFAT5 KO mice with LCMV infection and ChIP at IFNB1; myeloid-specific KO with co-IP of NF-κB/AP-1/p300 complex in microglia

    PMID:31816635 PMID:33292328

    Open questions at the time
    • How NFAT5 is directed to activating vs. repressive roles at different loci is undefined
    • Full spectrum of innate immune genes regulated by NFAT5 not mapped
  15. 2021 High

    NFAT5 was discovered to recognize R-loop structures via its RHD, recruit METTL3 for m6A-dependent R-loop resolution and RNaseH1 loading, with PARP1-mediated PARylation directing NFAT5 to damage-associated R-loops — establishing a genome integrity function independent of classical transcription.

    Evidence Single-molecule imaging of R-loop binding, co-IP of METTL3 and PARP1, PARylation assays, R-loop quantification by S9.6 antibody

    PMID:33313823 PMID:34049076

    Open questions at the time
    • Whether NFAT5 encircles R-loops as it does dsDNA is structurally unresolved
    • Relative contribution of NFAT5 vs. other R-loop sensors to genome stability not quantified
  16. 2023 High

    NFAT5 was found to promote CD8+ T cell exhaustion selectively in the hyperosmolar tumor microenvironment by driving TOX and PD-1 expression, while EGFR-activated EZH2 methylation at K668 was shown to stabilize NFAT5 protein by blocking TRAF6-mediated ubiquitination and lysosomal degradation — revealing tumor-specific regulatory mechanisms.

    Evidence NFAT5 gain/loss-of-function in tumor-bearing vs. LCMV-infected mice; co-IP/ubiquitination assays with K668 mutagenesis, xenograft and PDX models

    PMID:37429858 PMID:37709986

    Open questions at the time
    • Whether EZH2-K668 methylation also operates outside tumor contexts is unknown
    • Therapeutic targetability of NFAT5 in tumor immunology not established

Open questions

Synthesis pass · forward-looking unresolved questions
  • Key unresolved questions include: the identity of the proximal osmosensor upstream of NFAT5, the structural basis of R-loop recognition by the RHD, how NFAT5 is differentially directed to activating versus repressive chromatin complexes at distinct loci, and whether the R-loop resolution function is coordinated with or independent of its transcriptional roles.
  • No proximal osmosensor identified
  • No structural model for NFAT5-R-loop interaction
  • Mechanism selecting coactivation vs. repression at individual loci unknown

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140110 transcription regulator activity 9 GO:0003677 DNA binding 5 GO:0003723 RNA binding 1
Localization
GO:0005634 nucleus 5 GO:0005654 nucleoplasm 3 GO:0005694 chromosome 2
Pathway
R-HSA-74160 Gene expression (Transcription) 9 R-HSA-162582 Signal Transduction 6 R-HSA-168256 Immune System 6 R-HSA-4839726 Chromatin organization 2 R-HSA-73894 DNA Repair 2 R-HSA-8953897 Cellular responses to stimuli 2 R-HSA-8953854 Metabolism of RNA 1
Partners
ABL1CDK5METTL3PARP1PLCG1PTPN6RELAWWTR1
Complex memberships
AP-1/NF-κB/p300 complexNF-κB enhanceosome (p65/p300 cofactor complex)

Evidence

Reading pass · 42 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1999 NFAT5 contains a Rel homology domain, binds DNA sequences similar to those recognized by NFAT1-4, but lacks the majority of Fos/Jun contact residues and does not bind cooperatively with Fos and Jun to DNA. Unlike NFAT1-4, NFAT5 is constitutively nuclear and its nuclear localization is not regulated by calcineurin-mediated dephosphorylation. DNA binding assays, co-transfection, nuclear/cytoplasmic fractionation, sequence analysis Proceedings of the National Academy of Sciences of the United States of America High 10377394
2002 TonEBP/NFAT5 adopts an NF-κB-like structure upon binding DNA and exists as a homodimer that completely encircles its asymmetric TonE DNA target, providing increased kinetic stability of the TonEBP-DNA complex. Crystal structure determination, biochemical DNA-encirclement assays, electrophoretic mobility shift assay Nature structural biology High 11780147
2002 The C-terminal transactivation domain (TAD) of TonEBP/OREBP has activity that varies directly with extracellular NaCl concentration, and tonicity-dependent regulation of this TAD involves posttranslational phosphorylation. Tyrosine kinase and CK2 inhibitors reduce TAD activity at high osmolality. GAL4-TAD chimera reporter assays, phosphorylation analysis, pharmacological inhibitors in transfected HepG2 cells Proceedings of the National Academy of Sciences of the United States of America High 11792870
2000 TonEBP/TonE pathway mediates tonicity-responsive transcriptional regulation of urea transporter UT-A1, UT-A3, and UT-A4; a TonE element at -377 bp of the UT-A promoter is required, and dominant-negative TonEBP abolishes hypertonic induction of UT-A expression. Luciferase reporter assay, EMSA supershift, dominant-negative co-transfection, deletion/mutation analysis The Journal of biological chemistry High 10995747
2006 Nuclear export of TonEBP/OREBP under isotonic conditions is mediated by a CRM1-dependent leucine-rich nuclear export sequence (NES) in the N-terminus; disruption of the NES causes constitutive nuclear accumulation. An auxiliary export domain distinct from the NES is required for hypotonicity-induced nuclear export. CRM1 physically interacts with TonEBP. GFP fusion imaging, site-directed mutagenesis, bimolecular fluorescence complementation (BiFC), immunocytochemistry, CRM1 inhibitor (leptomycin B) The Journal of biological chemistry High 16782704
2010 c-Abl kinase directly phosphorylates TonEBP/OREBP at tyrosine-143 in response to high NaCl, contributing to its nuclear localization and transactivating activity. c-Abl co-immunoprecipitates with TonEBP, and inhibition of c-Abl (imatinib, siRNA, kinase-dead mutant) drastically reduces TonEBP nuclear location and transactivating activity. Co-immunoprecipitation, in vitro kinase assay, siRNA knockdown, imatinib pharmacological inhibition, phosphorylation-site mutagenesis (Y143A) FASEB journal High 20585028
2009 PLC-γ1 is recruited to TonEBP/OREBP at phospho-Y143 in response to high NaCl; this interaction is required for TonEBP nuclear localization and transactivating activity. PLC-γ1 co-immunoprecipitates with wild-type but not Y143A TonEBP, and is present in the protein complex on TonEBP DNA binding sites. Co-immunoprecipitation, siRNA knockdown, PLC-γ1-null cells, reporter assay, nuclear localization quantification, EMSA Proceedings of the National Academy of Sciences of the United States of America High 20080774
2011 Rac1 and its scaffold OSM support TonEBP/OREBP transcriptional and transactivating activity through PLC-γ1, not through p38 MAPK. In PLC-γ1-null cells, catalytically active Rac1 cannot increase TonEBP activity unless PLC-γ1 is reconstituted. siRNA knockdown of Rac1/OSM/MKK3/MKK6, PLC-γ1-null mouse embryonic fibroblasts, reconstitution, dominant-negative constructs, reporter assay Proceedings of the National Academy of Sciences of the United States of America High 21712438
2010 SHP-1 protein tyrosine phosphatase dephosphorylates TonEBP/OREBP at Y143, reducing both its nuclear localization and transactivating activity. High NaCl inhibits SHP-1 by phosphorylating it at Ser591, thereby promoting TonEBP activation. SHP-1 co-immunoprecipitates with TonEBP. Genome-wide phosphatase siRNA screen, SHP-1 overexpression, co-immunoprecipitation, in vivo and in vitro dephosphorylation assays Proceedings of the National Academy of Sciences of the United States of America High 20351292
2011 CDK5 is activated by high NaCl and directly phosphorylates TonEBP/OREBP at threonine-135, which accelerates rapid nuclear localization of TonEBP but does not affect its transactivating activity. CDK5 inhibition reduces the early increase in TonEBP nuclear localization. Mass spectrometry of phosphorylation sites, alanine mutagenesis, CDK5 activity assay, siRNA knockdown, CDK5 inhibitor, nuclear localization quantification Molecular biology of the cell High 21209322
2006 TonEBP/OREBP associates with multiple nuclear proteins including Hsp90β, PARP-1, DNA-PK, RNA helicases, and hnRNPs in response to high NaCl. Hsp90 enhances TonEBP transcriptional and transactivating activity and sustains increased TonEBP protein abundance, while PARP-1 reduces TonEBP transcriptional activity. Stable expression of N-terminal TonEBP fragment, nuclear immunoprecipitation followed by mass spectrometry, Western blot confirmation, EMSA complex analysis, functional reporter assay American journal of physiology. Renal physiology High 17148781
2010 TonEBP interacts with NF-κB p65 in a tonicity-dependent manner; NF-κB-TonEBP complexes bind κB elements and enhance NF-κB activity. This is dependent on Akt signaling activated by hypertonicity. TonEBP does not affect IκBα degradation or p65 nuclear translocation. Co-immunoprecipitation, chromatin immunoprecipitation, reporter assay, Akt pathway inhibitors, siRNA knockdown in renal epithelial cells and macrophages Molecular biology of the cell High 20685965
2016 TonEBP is required for recruitment of p300 co-activator to the LPS-induced NF-κB enhanceosome; TonEBP itself does not bind DNA in this context but acts as a cofactor. Recombinant TonEBP unable to recruit p300 does not stimulate NF-κB. Myeloid-specific TonEBP deletion results in milder sepsis and inflammation. ChIP, co-immunoprecipitation, dominant-negative/mutant TonEBP, myeloid-specific knockout mice, reporter assay, sepsis model Scientific reports High 27118681
2021 TonEBP recognizes R-loops (RNA-DNA hybrid structures) generated by DNA-damaging agents and recruits METTL3 to R-loops through its Rel homology domain (RHD) for m6A RNA methylation-mediated R-loop resolution. TonEBP also recruits RNaseH1 to R-loops via METTL3 interaction. Single-molecule imaging shows TonEBP preferentially binds R-loops via 3D collision and 1D diffusion along DNA. Single-molecule imaging, biochemical binding assays, co-immunoprecipitation, siRNA knockdown, R-loop quantification (S9.6 antibody), cell survival assay Nucleic acids research High 33313823
2021 PARP1-mediated PARylation of TonEBP is required for recruitment of TonEBP to sites of R-loop-associated DNA damage (camptothecin-induced). TonEBP interacts with PARP1 and loss of TonEBP increases R-loop accumulation and DNA damage. Co-immunoprecipitation, biotin-switch/PARylation assay, siRNA knockdown, R-loop quantification, cell survival assay DNA repair High 34049076
2005 High NaCl stabilizes TonEBP/OREBP mRNA through its 5'-UTR (not the 3'-UTR), contributing to the increase in TonEBP mRNA and protein abundance. mRNA half-life measured after actinomycin D block shows 6 h stabilization. Actinomycin D mRNA stability assay, luciferase reporters with UTR constructs, quantitative northern/western analysis American journal of physiology. Renal physiology High 15900024
2005 High NaCl increases reactive oxygen species (ROS), specifically superoxide from mitochondria (complexes I and III), which contribute to TonEBP/OREBP activation by increasing its transactivating activity but not its nuclear translocation. ROS inhibitors (NAC, MnTBAP, rotenone, myxothiazol), reporter assay, mitochondrial complex inhibitors, BGT1 mRNA measurement American journal of physiology. Renal physiology High 15769933 16303854
2012 TAZ physically interacts with NFAT5 in response to hyperosmotic stress via c-Abl-mediated phosphorylation of TAZ at tyrosine-316, and phosphorylated TAZ suppresses NFAT5 DNA binding and transcriptional activity. TAZ-deficient cells show increased NFAT5 activity, reversed by wild-type but not Y316F mutant TAZ. Co-immunoprecipitation, site-directed mutagenesis (Y316F), TAZ knockdown/knockout, reporter assay, EMSA Molecular and cellular biology High 23045390
2000 NFAT5/NFATL1 induction upon T cell activation is inhibited by cyclosporin A and FK506 (calcineurin inhibitors), and constitutively active calcineurin induces NFATL1-dependent transcription, placing NFAT5 induction downstream of calcineurin in T cell receptor signaling. Hyperosmotic induction of NFATL1 is calcineurin-independent. Primary T cell activation, pharmacological calcineurin inhibition, constitutively active calcineurin overexpression, dominant-negative NFATc1, reporter assay Journal of immunology Medium 11046013
2013 IKKβ regulates NFAT5 expression in thymocytes; NFAT5 promotes survival of αβ T cell receptor thymocytes at the β-selection checkpoint by upregulating prosurvival factors A1 and Bcl2 and attenuating the p53/Noxa proapoptotic axis, in an osmostress-independent manner. Conditional NFAT5 knockout mice, IKKβ genetic manipulation, gene expression analysis, apoptosis assay, allelic exclusion assessment Proceedings of the National Academy of Sciences of the United States of America High 24043824
2007 Hypertonic activation of ERK1/2 signaling regulates the transactivation domain (TAD) of TonEBP/OREBP in nucleus pulposus cells; dominant-negative ERK suppresses TonEBP TAD activity while constitutively active MEK1 and wild-type ERK enhance it. EMSA, luciferase reporter assay, dominant-negative and constitutively active kinase constructs, ERK siRNA, inhibitor treatment Journal of bone and mineral research Medium 17371162
2008 p38α and p38δ isoforms have opposing effects on TonEBP/OREBP activity when both are activated by high NaCl: p38α overexpression increases TonEBP activity while p38δ overexpression and siRNA knockdown of p38δ both reduce and enhance TonEBP activity respectively. MKP-1 phosphatase inhibits p38 but has no net effect on TonEBP because of these opposing actions. p38 isoform-specific siRNA, dominant-negative MKP-1, reporter assay, phosphorylation analysis Proceedings of the National Academy of Sciences of the United States of America Medium 18367666
2010 TonEBP is required for optimal expression of AQP1 in renal inner medulla under hypertonic conditions; a TonEBP binding site 811 bp upstream of the AQP1 exon is essential, and site-directed mutation of this TonE site reduces AQP1 luciferase activity by 54%. TonEBP mutant mice show markedly decreased AQP1 in the inner medulla. shRNA knockdown, rottlerin inhibitor, luciferase reporter with TonE-site mutation, TonEBP mutant mice The Journal of biological chemistry High 20639513
2019 TonEBP suppresses the β3-adrenoreceptor gene promoter in subcutaneous adipocytes by recruiting DNMT1 DNA methylase and methylating the promoter, thereby epigenetically repressing thermogenesis. Adipocyte-specific TonEBP deficiency results in resistance to high-fat diet-induced obesity. ChIP, DNA methylation (bisulfite sequencing), adipocyte-specific knockout mice, promoter reporter assay, gene expression analysis Nature communications High 31387996
2015 TonEBP binds the PPARγ2 promoter and blocks epigenetic transition (chromatin remodeling) required for PPARγ2 activation, thereby suppressing adipogenesis and insulin signaling. Reduction of TonEBP expression leads to enhanced epigenetic transition, PPARγ2 expression, adipogenesis, and insulin response. ChIP, chromatin accessibility assay, reporter assay, TonEBP knockdown/overexpression Scientific reports Medium 26042523
2019 TonEBP suppresses HO-1 expression by blocking Nrf2 binding to the HO-1 promoter, thereby promoting M1 macrophage polarization. TonEBP knockdown releases this suppression, allowing Nrf2-driven HO-1 expression. ChIP, reporter assay, TonEBP knockdown, Nrf2 binding competition assay Frontiers in immunology Medium 31057560
2020 NFAT5 represses TLR3- and virus-induced IFNβ expression in macrophages and dendritic cells by binding an evolutionarily conserved sequence in the IFNB1 enhanceosome that overlaps a key IRF3 binding site, opposing IRF3-mediated IFN-I induction. NFAT5 knockout mice, LCMV infection model, poly(I:C) challenge, ChIP, reporter assay, EMSA The Journal of experimental medicine High 31816635
2023 NFAT5 transcriptional activity is selectively activated in the tumor microenvironment by hyperosmolarity; NFAT5 promotes exhaustion of CD8+ T cells (reducing tumor control) by enhancing expression of TOX and PD-1 in precursor exhausted T cells, but this function is not required during chronic viral infection where osmolarity is normal. NFAT5 overexpression and deletion in CD8+ T cells, tumor models, chronic LCMV infection, flow cytometry for exhaustion markers, cytokine production assays Nature immunology High 37709986
2023 EGFR activation induces phosphorylated EZH2 (Ser21) to bind and methylate NFAT5 at K668; this methylation prevents TRAF6-mediated K63-linked ubiquitination and lysosomal degradation of NFAT5 in the cytoplasm, causing NFAT5 protein stabilization, nuclear accumulation, and activation with upregulation of MGMT transcription. Co-immunoprecipitation, ubiquitination assays, mutagenesis (K668), lysosomal degradation assays, ChIP, in vivo xenograft models, PDX models Nature communications High 37429858
2014 Biomechanical stretch activates NFAT5 nuclear translocation in vascular smooth muscle cells (VSMCs) through c-Jun N-terminal kinase (JNK) for protein abundance regulation and palmitoyltransferases for nuclear translocation; NFAT5 then transcriptionally upregulates tenascin-C, stimulating VSMC migration. Immunofluorescence, Western blot, ChIP, DNA microarray, cyclic stretch device, pharmacological inhibitors, isolated artery perfusion model Journal of the American Heart Association Medium 24614757
2009 In nucleus pulposus cells, NFAT5/TonEBP controls AQP2 expression in response to both osmolarity and intracellular calcium, independently of the calcineurin-NFAT1-4 pathway. TonEBP-null MEFs confirm TonEBP requirement, and mutant TonE reporter constructs show direct promoter binding is necessary. AQP2 reporter assay, TonEBP-null MEFs, calcineurin null MEFs, calcium ionophore treatment, NFAT1-4 co-transfection, mutant TonE elements Journal of bone and mineral research Medium 19138132
2009 Hypertonic stress induces rapid and reversible nucleosome depletion around TonE/ORE binding sites of the aldose reductase gene in an OREBP-independent initial step; OREBP potentiates this nucleosome loss and drives OREBP-dependent histone hyperacetylation spanning 5' upstream sequences and exons. Chromatin immunoprecipitation, nucleosome positioning assay, histone acetylation ChIP, OREBP-null comparison PloS one Medium 20041176
2017 NFAT5 promotes macrophage survival in rheumatoid arthritis by inducing CCL2 secretion; CCL2 acts in an autocrine/paracrine manner to confer apoptotic resistance. In vivo, NFAT5-deficient macrophages show increased apoptosis and reduced joint destruction, and intra-articular injection of recombinant CCL2 reverses the protective effect of NFAT5 haploinsufficiency. NFAT5-deficient macrophages, apoptosis assays, recombinant CCL2 rescue, intra-articular injection in Nfat5+/- mice, transcriptome profiling The Journal of clinical investigation High 28192374
2014 PKC-α contributes to high NaCl-induced activation of NFAT5 through ERK1/2 signaling (but not through SHP-1-S591); PKC-α knockout reduces NFAT5 protein abundance and target gene expression in renal inner medulla. PKC-α knockout mice, ERK1/2 siRNA knockdown, combined knockdown epistasis, reporter assay, inner medulla gene expression American journal of physiology. Renal physiology Medium 25391900
2010 NFAT5 regulates T lymphocyte homeostasis and CD24 expression under hypernatremia; NFAT5 binds the Cd24 promoter in response to hypertonicity, facilitates local chromatin derepression, and enhances CD24 mRNA and protein expression; CD24 is required for T cell expansion under osmostress. NFAT5-null and T cell-specific KO mice, ChIP, chromatin accessibility assay, in vivo hypernatremia model, CD24 rescue experiments Journal of immunology High 21037089
2019 NFAT5 directly controls AQP4 expression in spinal dorsal horn astrocytes. Aurora kinase B (AURKB) phosphorylates NFAT5, enhancing its protein stability and nuclear translocation, thereby regulating AQP4 expression and astrocyte swelling in neuropathic pain. In vivo knockdown/knockout, ChIP for NFAT5 at AQP4 promoter, AURKB inhibition, phosphorylation analysis, astrocyte swelling assay Advanced science Medium 38195869
2012 NFAT5 regulates the canonical Wnt pathway during cardiomyogenesis by coordinating expression of Wnt ligands (Wnt3, Wnt3a) and antagonists (Dkk1, Cerberus1); inhibition of NFAT5 blocks mesodermal differentiation and Wnt signaling activation, which is rescued by pharmacological Wnt activation. Dominant-negative NFAT5, TOPflash Wnt reporter assay, BIO pharmacological rescue, gene expression analysis in P19CL6 cells Biochemical and biophysical research communications Medium 22935419
2012 In intervertebral disc nucleus pulposus cells, pharmacological inhibition of Cdc42 and Arp2/3 prevents TonEBP/NFAT5 from recruiting cofactors in response to hyperosmolarity challenge, linking actin cytoskeletal dynamics to TonEBP-mediated osmoadaptation. Pharmacological Cdc42/Arp2/3 inhibitors, Arpc2 conditional knockout mice, cofactor recruitment assay JCI insight Medium 31961823
2020 In microglia, TonEBP acts as a transcriptional cofactor for both NF-κB and AP-1, promoting LPS-induced pro-inflammatory gene expression; myeloid-specific TonEBP deletion blocks microglial activation and neuronal cell death in vivo. The natural molecule cerulenin disrupts the TonEBP/NF-κB/AP-1/p300 complex. shRNA knockdown, myeloid-specific TonEBP knockout mice, AP-1 reporter assay, co-immunoprecipitation of TonEBP/NF-κB/AP-1/p300 complex, cerulenin pharmacology, LPS brain injection model Journal of neuroinflammation Medium 33292328
2016 NFAT5 directly activates CACNA1C (L-type calcium channel) transcription by binding the consensus sequence TGGAAGCGTTC in the CACNA1C promoter; NFAT5 knockdown suppresses CACNA1C expression and L-type calcium current in cardiomyocytes, and morpholino knockdown in zebrafish causes non-contractile ventricle rescued by CACNA1C or NFAT5 overexpression. Comparative genomics, ChIP, reporter assay, siRNA knockdown, electrophysiology (L-type Ca2+ current), morpholino knockdown in zebrafish, rescue overexpression Journal of molecular medicine Medium 27368804
2014 NFAT5 acts synergistically with TNF-α to induce CXCL1, CXCL2, and CXCL3 expression in nucleus pulposus cells through a conserved NF-κB-binding site (not a TonE site), indicating cross-talk between TonEBP and NF-κB Rel family members at CXCL gene promoters. RNA sequencing, CXCL promoter luciferase reporter with NF-κB site mutations, TonEBP knockdown, ChIP The Journal of biological chemistry Medium 28842479
2018 NFAT5 upregulates NFAT5 binds the Rnf183 promoter at a conserved TonE motif and activates Rnf183 transcription specifically under hypertonic conditions; ChIP shows enhanced NFAT5 binding at this site in hypertonic stress, and mutation of the TonE site abolishes hypertonic induction. siRNA knockdown of NFAT5, luciferase reporter with TonE-site mutation, ChIP, gene expression analysis The Journal of biological chemistry Medium 30413537

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1999 NFAT5, a constitutively nuclear NFAT protein that does not cooperate with Fos and Jun. Proceedings of the National Academy of Sciences of the United States of America 318 10377394
2000 The NFAT-related protein NFATL1 (TonEBP/NFAT5) is induced upon T cell activation in a calcineurin-dependent manner. Journal of immunology (Baltimore, Md. : 1950) 137 11046013
2000 The TonE/TonEBP pathway mediates tonicity-responsive regulation of UT-A urea transporter expression. The Journal of biological chemistry 119 10995747
2002 Activity of the TonEBP/OREBP transactivation domain varies directly with extracellular NaCl concentration. Proceedings of the National Academy of Sciences of the United States of America 118 11792870
2002 Structure of a TonEBP-DNA complex reveals DNA encircled by a transcription factor. Nature structural biology 104 11780147
2014 Extracellular osmolarity regulates matrix homeostasis in the intervertebral disc and articular cartilage: evolving role of TonEBP. Matrix biology : journal of the International Society for Matrix Biology 96 25172826
2019 Role of NFAT5 in the Immune System and Pathogenesis of Autoimmune Diseases. Frontiers in immunology 90 30873159
2011 Tonicity-independent regulation of the osmosensitive transcription factor TonEBP (NFAT5). American journal of physiology. Cell physiology 89 21998140
2010 Osmoprotective transcription factor NFAT5/TonEBP modulates nuclear factor-kappaB activity. Molecular biology of the cell 88 20685965
2002 The osmoprotective function of the NFAT5 transcription factor in T cell development and activation. Journal of immunology (Baltimore, Md. : 1950) 86 12421923
2007 MEK/ERK signaling controls osmoregulation of nucleus pulposus cells of the intervertebral disc by transactivation of TonEBP/OREBP. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research 85 17371162
2019 CircFOXO3 promotes glioblastoma progression by acting as a competing endogenous RNA for NFAT5. Neuro-oncology 83 31504797
2006 How tonicity regulates genes: story of TonEBP transcriptional activator. Acta physiologica (Oxford, England) 80 16734761
2004 Osmotic response element-binding protein (OREBP) is an essential regulator of the urine concentrating mechanism. The Journal of biological chemistry 77 15347663
2021 TonEBP recognizes R-loops and initiates m6A RNA methylation for R-loop resolution. Nucleic acids research 73 33313823
2010 Tonicity-responsive microRNAs contribute to the maximal induction of osmoregulatory transcription factor OREBP in response to high-NaCl hypertonicity. Nucleic acids research 73 20852262
2017 Transcription factor NFAT5 promotes macrophage survival in rheumatoid arthritis. The Journal of clinical investigation 72 28192374
2020 The evolving role of TonEBP as an immunometabolic stress protein. Nature reviews. Nephrology 65 32157251
2005 High NaCl increases TonEBP/OREBP mRNA and protein by stabilizing its mRNA. American journal of physiology. Renal physiology 61 15900024
2017 Upregulation of DARS2 by HBV promotes hepatocarcinogenesis through the miR-30e-5p/MAPK/NFAT5 pathway. Journal of experimental & clinical cancer research : CR 60 29052520
2004 Transcription factor tonicity-responsive enhancer-binding protein (TonEBP) which transactivates osmoprotective genes is expressed and upregulated following acute systemic hypertonicity in neurons in brain. Neuroscience 59 14960342
2003 Silencing of TonEBP/NFAT5 transcriptional activator by RNA interference. Journal of the American Society of Nephrology : JASN 57 12538727
2003 The role of NFAT5/TonEBP in establishing an optimal intracellular environment. Archives of biochemistry and biophysics 57 12729611
2014 NFAT5-mediated expression of S100A4 contributes to proliferation and migration of renal carcinoma cells. Frontiers in physiology 56 25152734
2006 Regulation of nucleocytoplasmic trafficking of transcription factor OREBP/TonEBP/NFAT5. The Journal of biological chemistry 56 16782704
2005 Increased reactive oxygen species contribute to high NaCl-induced activation of the osmoregulatory transcription factor TonEBP/OREBP. American journal of physiology. Renal physiology 55 15769933
2019 Regulation of Inflammatory Functions of Macrophages and T Lymphocytes by NFAT5. Frontiers in immunology 54 30949179
2022 NLRC3 expression in macrophage impairs glycolysis and host immune defense by modulating the NF-κB-NFAT5 complex during septic immunosuppression. Molecular therapy : the journal of the American Society of Gene Therapy 53 36068919
2016 LPS-induced NFκB enhanceosome requires TonEBP/NFAT5 without DNA binding. Scientific reports 53 27118681
2011 TonEBP stimulates multiple cellular pathways for adaptation to hypertonic stress: organic osmolyte-dependent and -independent pathways. American journal of physiology. Renal physiology 52 21209002
2010 The expression of aquaporin-1 in the medulla of the kidney is dependent on the transcription factor associated with hypertonicity, TonEBP. The Journal of biological chemistry 52 20639513
2018 A miRNA181a/NFAT5 axis links impaired T cell tolerance induction with autoimmune type 1 diabetes. Science translational medicine 51 29298866
2023 Activation of the transcription factor NFAT5 in the tumor microenvironment enforces CD8+ T cell exhaustion. Nature immunology 49 37709986
2019 HIF1A and NFAT5 coordinate Na+-boosted antibacterial defense via enhanced autophagy and autolysosomal targeting. Autophagy 49 30982460
2005 Mitochondrial reactive oxygen species contribute to high NaCl-induced activation of the transcription factor TonEBP/OREBP. American journal of physiology. Renal physiology 49 16303854
2012 NFAT5 expression in bone marrow-derived cells enhances atherosclerosis and drives macrophage migration. Frontiers in physiology 48 22934063
2017 NFAT5-Regulated Macrophage Polarization Supports the Proinflammatory Function of Macrophages and T Lymphocytes. Journal of immunology (Baltimore, Md. : 1950) 47 29150563
2010 NFAT5 regulates T lymphocyte homeostasis and CD24-dependent T cell expansion under pathologic hypernatremia. Journal of immunology (Baltimore, Md. : 1950) 45 21037089
2023 Macrophage transcription factor TonEBP promotes systemic lupus erythematosus and kidney injury via damage-induced signaling pathways. Kidney international 44 37088425
2017 TNF-α promotes nuclear enrichment of the transcription factor TonEBP/NFAT5 to selectively control inflammatory but not osmoregulatory responses in nucleus pulposus cells. The Journal of biological chemistry 43 28842479
2015 NFAT5 promotes proliferation and migration of lung adenocarcinoma cells in part through regulating AQP5 expression. Biochemical and biophysical research communications 43 26299924
2002 Adaptation of kidney medulla to hypertonicity: role of the transcription factor TonEBP. International review of cytology 43 11952228
2014 Arterial wall stress controls NFAT5 activity in vascular smooth muscle cells. Journal of the American Heart Association 42 24614757
2019 TonEBP/NFAT5 promotes obesity and insulin resistance by epigenetic suppression of white adipose tissue beiging. Nature communications 41 31387996
2009 Osmolarity and intracellular calcium regulate aquaporin2 expression through TonEBP in nucleus pulposus cells of the intervertebral disc. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research 41 19138132
2011 Src kinase pathway is involved in NFAT5-mediated S100A4 induction by hyperosmotic stress in colon cancer cells. American journal of physiology. Cell physiology 40 21289293
2014 The xanthine oxidase-NFAT5 pathway regulates macrophage activation and TLR-induced inflammatory arthritis. European journal of immunology 38 25044064
2012 TAZ suppresses NFAT5 activity through tyrosine phosphorylation. Molecular and cellular biology 38 23045390
2006 Proteomic identification of proteins associated with the osmoregulatory transcription factor TonEBP/OREBP: functional effects of Hsp90 and PARP-1. American journal of physiology. Renal physiology 38 17148781
2012 NFAT5 is activated by hypoxia: role in ischemia and reperfusion in the rat kidney. PloS one 37 22768306
2004 Maturation of TonEBP expression in developing rat kidney. American journal of physiology. Renal physiology 36 15226152
2009 Phospholipase C-gamma1 is involved in signaling the activation by high NaCl of the osmoprotective transcription factor TonEBP/OREBP. Proceedings of the National Academy of Sciences of the United States of America 35 20080774
2017 Suppression of NFAT5-mediated Inflammation and Chronic Arthritis by Novel κB-binding Inhibitors. EBioMedicine 34 28396011
2009 Hypertonic induction of COX2 expression requires TonEBP/NFAT5 in renal epithelial cells. Biochemical and biophysical research communications 34 19146830
2019 MiR-361-3p/Nfat5 Signaling Axis Controls Cementoblast Differentiation. Journal of dental research 33 31343932
2011 Rac1/osmosensing scaffold for MEKK3 contributes via phospholipase C-gamma1 to activation of the osmoprotective transcription factor NFAT5. Proceedings of the National Academy of Sciences of the United States of America 33 21712438
2008 MKP-1 inhibits high NaCl-induced activation of p38 but does not inhibit the activation of TonEBP/OREBP: opposite roles of p38alpha and p38delta. Proceedings of the National Academy of Sciences of the United States of America 33 18367666
2010 c-Abl mediates high NaCl-induced phosphorylation and activation of the transcription factor TonEBP/OREBP. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 32 20585028
2012 NFAT5 contributes to osmolality-induced MCP-1 expression in mesothelial cells. Mediators of inflammation 31 22619484
2011 High NaCl-induced activation of CDK5 increases phosphorylation of the osmoprotective transcription factor TonEBP/OREBP at threonine 135, which contributes to its rapid nuclear localization. Molecular biology of the cell 31 21209322
2008 Interstitial tonicity controls TonEBP expression in the renal medulla. Kidney international 31 19052532
2016 Context-dependent regulation of Th17-associated genes and IFNγ expression by the transcription factor NFAT5. Immunology and cell biology 30 27479742
2001 Amino acid depletion activates TonEBP and sodium-coupled inositol transport. American journal of physiology. Cell physiology 30 11350742
2019 miR-194 suppresses high glucose-induced non-small cell lung cancer cell progression by targeting NFAT5. Thoracic cancer 29 30900402
2013 NFAT5 induction by the pre-T-cell receptor serves as a selective survival signal in T-lymphocyte development. Proceedings of the National Academy of Sciences of the United States of America 28 24043824
2020 Arp2/3 inactivation causes intervertebral disc and cartilage degeneration with dysregulated TonEBP-mediated osmoadaptation. JCI insight 26 31961823
2016 Inflammatory signals induce the expression of tonicity-responsive enhancer binding protein (TonEBP) in microglia. Journal of neuroimmunology 26 27235345
2010 Contribution of SHP-1 protein tyrosine phosphatase to osmotic regulation of the transcription factor TonEBP/OREBP. Proceedings of the National Academy of Sciences of the United States of America 26 20351292
2006 Large discrepancies in cellular distribution of the tonicity-induced expression of osmoprotective genes and their regulatory transcription factor TonEBP in rat brain. Neuroscience 26 16890372
2006 Sequential expression of NKCC2, TonEBP, aldose reductase, and urea transporter-A in developing mouse kidney. American journal of physiology. Renal physiology 26 16926446
2009 Inducible nucleosome depletion at OREBP-binding-sites by hypertonic stress. PloS one 25 20041176
2020 The transcription factor NFAT5 limits infection-induced type I interferon responses. The Journal of experimental medicine 24 31816635
2020 NFAT5 promotes oral squamous cell carcinoma progression in a hyperosmotic environment. Laboratory investigation; a journal of technical methods and pathology 24 32901097
2014 PKC-α contributes to high NaCl-induced activation of NFAT5 (TonEBP/OREBP) through MAPK ERK1/2. American journal of physiology. Renal physiology 24 25391900
2024 Inhibition of NFAT5-Dependent Astrocyte Swelling Alleviates Neuropathic Pain. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 23 38195869
2023 Exosomal MFI2-AS1 sponge miR-107 promotes non-small cell lung cancer progression through NFAT5. Cancer cell international 23 36934264
2021 PARP1-mediated PARylation of TonEBP prevents R-loop-associated DNA damage. DNA repair 23 34049076
2023 Lysine methylation promotes NFAT5 activation and determines temozolomide efficacy in glioblastoma. Nature communications 22 37429858
2020 Potential Role of Gene Regulator NFAT5 in the Pathogenesis of Diabetes Mellitus. Journal of diabetes research 22 33015193
2009 Expression and function of NFAT5 in medullary thick ascending limb (mTAL) cells. American journal of physiology. Renal physiology 22 19369291
2017 NFAT5-sensitive Orai1 expression and store-operated Ca2+ entry in megakaryocytes. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 21 28446591
2012 NFAT5 regulates the canonical Wnt pathway and is required for cardiomyogenic differentiation. Biochemical and biophysical research communications 21 22935419
2007 Downregulation of renal TonEBP in hypokalemic rats. American journal of physiology. Renal physiology 21 17409277
2001 Urea inhibits hypertonicity-inducible TonEBP expression and action. American journal of physiology. Renal physiology 21 11292634
2016 NFAT5-mediated CACNA1C expression is critical for cardiac electrophysiological development and maturation. Journal of molecular medicine (Berlin, Germany) 20 27368804
2009 Brx shines a light on the route from hyperosmolarity to NFAT5. Science signaling 20 19351952
2024 Serum amyloid A expression in liver promotes synovial macrophage activation and chronic arthritis via NFAT5. The Journal of clinical investigation 19 38426494
2020 Genistein-Calcitriol Mitigates Hyperosmotic Stress-Induced TonEBP, CFTR Dysfunction, VDR Degradation and Inflammation in Dry Eye Disease. Clinical and translational science 19 32896986
2019 NFAT5/TonEBP controls early acquisition of notochord phenotypic markers, collagen composition, and sonic hedgehog signaling during mouse intervertebral disc embryogenesis. Developmental biology 19 31301300
2012 Sepsis-induced urinary concentration defect is related to nitric oxide-dependent inactivation of TonEBP/NFAT5, which downregulates renal medullary solute transport proteins and aquaporin-2. Critical care medicine 19 22610191
2002 Rottlerin inhibits tonicity-dependent expression and action of TonEBP in a PKCdelta-independent fashion. American journal of physiology. Renal physiology 19 11880333
2020 TonEBP in dendritic cells mediates pro-inflammatory maturation and Th1/Th17 responses. Cell death & disease 18 32499518
2019 NFAT5 is Regulated by p53/miR-27a Signal Axis and Promotes Mouse Ovarian Granulosa Cells Proliferation. International journal of biological sciences 18 30745821
2019 TonEBP Suppresses the HO-1 Gene by Blocking Recruitment of Nrf2 to Its Promoter. Frontiers in immunology 18 31057560
2019 Osteoprotective action of low-salt diet requires myeloid cell-derived NFAT5. JCI insight 18 31801906
2018 Transcription Factor NFAT5 Promotes Migration and Invasion of Rheumatoid Synoviocytes via Coagulation Factor III and CCL2. Journal of immunology (Baltimore, Md. : 1950) 18 29794013
2018 NFAT5 up-regulates expression of the kidney-specific ubiquitin ligase gene Rnf183 under hypertonic conditions in inner-medullary collecting duct cells. The Journal of biological chemistry 18 30413537
2015 TonEBP suppresses adipogenesis and insulin sensitivity by blocking epigenetic transition of PPARγ2. Scientific reports 18 26042523
2014 TonEBP/NFAT5 regulates ACTBL2 expression in biomechanically activated vascular smooth muscle cells. Frontiers in physiology 18 25520667
2020 Microglial TonEBP mediates LPS-induced inflammation and memory loss as transcriptional cofactor for NF-κB and AP-1. Journal of neuroinflammation 17 33292328