Affinage

NKX2-5

Homeobox protein Nkx-2.5 · UniProt P52952

Length
324 aa
Mass
34.9 kDa
Annotated
2026-06-10
100 papers in source corpus 39 papers cited in narrative 39 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

NKX2-5 is a homeodomain transcription factor that operates as a central node of the cardiac gene regulatory network, acting in a gene-dosage-dependent manner to direct chamber and conduction-system identity, cardiomyocyte differentiation, and adult cardiomyocyte homeostasis (PMID:10021345, PMID:15085192, PMID:11889119). Its homeodomain mediates sequence-specific DNA binding to a TAAG/NKE consensus through the conserved Tyr54 residue, and also drives homodimerization (requiring Lys193/Arg194) and heterodimerization with related NK-2 factors (PMID:22849347, PMID:11042197). NKX2-5 nucleates combinatorial transcriptional complexes by physically associating with GATA4, TBX5, MEF2C, SRF, and the LIM protein Cal, yielding synergistic activation of cardiac promoters such as Nppa/ANF, cardiac alpha-actin, and Lrrc10 (PMID:9312027, PMID:11431700, PMID:19035347, PMID:11983708, PMID:23751912). Through these partnerships it sits genetically upstream of an extensive target set — activating myocardin, eHAND/Irx4, Id2, Rspo3, and HEY2 while repressing Isl1 and beta-catenin — thereby specifying the ventricular conduction system, promoting second heart field growth via Wnt/Rspo3 signaling, and balancing progenitor fate (PMID:14645532, PMID:11784028, PMID:17604724, PMID:25053429, PMID:29636455, PMID:25524439, PMID:19479054). NKX2-5 additionally recruits the CHD4/NuRD repressor to silencer elements to exclude noncardiac (skeletal and smooth muscle) gene programs from the developing heart (PMID:35450884). Pathogenic human homeodomain mutations cause loss of DNA binding and transcriptional activation, often acting dominant-negatively, and underlie congenital heart disease with conduction defects (PMID:10903346, PMID:11457872). Its activity is tuned by post-translational and post-transcriptional inputs: SUMOylation (at K51 plus a predominant cardiac site) stabilizes complexes and enhances activity, SIRT1 deacetylation at K182 within the homeodomain inhibits it by weakening SRF/TBX5 binding, and the RHAU/DHX36 helicase controls Nkx2-5 mRNA translation and decay via 5'-UTR G-quadruplex and 3'-UTR elements (PMID:18579533, PMID:27819261, PMID:26489465).

Mechanistic history

Synthesis pass · year-by-year structured walk · 16 steps
  1. 1997 High

    Established that NKX2-5 does not act alone but partners with GATA4 to synergistically activate cardiac promoters, defining the combinatorial logic of the cardiac transcription network.

    Evidence In vitro binding, Co-IP, and ANF promoter reporter assays with deletion mapping in heterologous cells

    PMID:9312027

    Open questions at the time
    • Did not resolve endogenous occupancy or genome-wide co-targets
    • GATA6 specificity mechanism not structurally defined
  2. 1999 High

    Placed Nkx2-5 genetically upstream of a battery of cardiac regulators and structural genes, establishing its role as a master regulator whose loss arrests heart development after looping.

    Evidence Nkx2-5 null mouse with in situ hybridization for 20 genes, TUNEL/PCNA, and chimera analysis

    PMID:10021345

    Open questions at the time
    • Direct vs. indirect targets not distinguished
    • Mechanism of dominant interfering effect in chimeras unresolved
  3. 2000 High

    Defined the molecular basis of NKX2-5-linked congenital heart disease — homeodomain mutations abolish DNA binding/activation and can act dominant-negatively through preserved dimerization.

    Evidence EMSA, luciferase reporters, homodimerization and nuclear localization assays on patient-derived mutants

    PMID:10903346 PMID:11042197

    Open questions at the time
    • Cell-type-dependent dominance (fibroblast vs. cardiomyocyte) not mechanistically explained
    • In vivo consequences of dimerization defects not tested here
  4. 2001 High

    Showed that NKX2-5 cooperates with TBX5 and dHAND on specific promoters and pathways, linking disease-causing TBX5 mutations to loss of synergy and defining genetic routes to ventricular formation.

    Evidence Y2H, Co-IP, GST pulldown, reporter assays, and double-null mouse epistasis

    PMID:11431700 PMID:11784028

    Open questions at the time
    • Structural basis of the NKX2-5–TBX5 interface not resolved
    • Direct target promoters at endogenous loci not mapped genome-wide
  5. 2002 High

    Demonstrated higher-order combinatorial complexes (with SRF and GATA4) and a postnatal requirement for NKX2-5 in cardiomyocyte survival, extending its role beyond development into adult homeostasis.

    Evidence Reporter assays in multiple cell lines, EMSA, dominant-negative/wild-type transgenic mice, and doxorubicin apoptosis assays

    PMID:11889119 PMID:11983708 PMID:14645532

    Open questions at the time
    • Stoichiometry of SRF/NKX2-5/GATA4 complexes unknown
    • Anti-apoptotic mechanism not connected to specific transcriptional targets
  6. 2004 High

    Identified gene-dosage as a quantitative determinant of conduction-system cell number and discovered the calcium-responsive cofactor Cal, linking NKX2-5 activity to physiological signaling.

    Evidence Nkx2-5 KO/heterozygous mouse cell counting with connexin IHC and electrophysiology; Y2H and Co-IP for Cal

    PMID:14757752 PMID:15085192

    Open questions at the time
    • Molecular trigger converting dosage into cell-number outcome unresolved
    • Cal nuclear shuttling signaling pathway not fully defined
  7. 2007 High

    Resolved NKX2-5 as a determinant of regional myocardial identity (pulmonary myocardium, conduction system) and showed it tunes pacemaking ion-channel programs.

    Evidence Hypomorphic and lineage-tracing mouse models; Id2 promoter reporters with compound haploinsufficiency; adenoviral overexpression in rat cardiomyocytes with patch clamp

    PMID:17250822 PMID:17498735 PMID:17604724 PMID:17823370

    Open questions at the time
    • Direct vs. indirect control of T-type/L-type Ca2+ channels not distinguished
    • Cell-autonomy mechanism for delayed Purkinje phenotype unresolved
  8. 2008 High

    Established NKX2-5 SUMOylation at K51 as an activity-enhancing, complex-stabilizing modification and showed cooperative control of ventricular differentiation with MEF2C, while inducible KO revealed postnatal regulation of ion-channel/Ca2+-handling genes.

    Evidence In vitro SUMOylation with PIAS E3 ligases and K51R mutagenesis; Co-IP/two-hybrid plus double-KO epistasis; tamoxifen-inducible conditional KO with RT-PCR/Western

    PMID:18579533 PMID:18689573 PMID:18722343 PMID:19035347

    Open questions at the time
    • In vivo physiological role of SUMOylation not established
    • Direct vs. indirect regulation of Nav1.5/RyR2 not separated
  9. 2011 High

    Connected NKX2-5 dosage to discrete fate decisions — SAN versus working myocardium, Isl1 repression, miR-1/Cdc42 signaling, and SHF Wnt/Rspo3 growth — defining how it partitions progenitor identity, and identified an additional dominant cardiac SUMOylation site.

    Evidence Transgenic gain-of-function, enhancer binding/reporter assays, compound heterozygous mouse and Drosophila genetics, inducible conditional KO, and systematic lysine mutagenesis

    PMID:21285290 PMID:21640717 PMID:21690310 PMID:21931855 PMID:25524439

    Open questions at the time
    • Identity of the predominant cardiac SUMOylation site remains undefined (Medium-confidence)
    • Direct enhancer targets of repression vs. indirect effects incompletely mapped
  10. 2012 High

    Provided atomic-resolution insight into DNA recognition, showing two homeodomains bind the ANF promoter independently with Tyr54 reading the TAAG motif.

    Evidence X-ray crystallography at 1.8 Å of the homeodomain–DNA complex

    PMID:22849347

    Open questions at the time
    • No structure of full-length protein or of cofactor complexes
    • Functional consequence of non-interacting dual-site binding in vivo untested
  11. 2014 High

    Established NKX2-5 as both an activator of growth-promoting targets (Rspo3, Lrrc10) and a suppressor of proliferation via Notch, clarifying its dual control of cardiomyocyte number.

    Evidence Conditional KO/rescue and pharmacological Wnt activation for Rspo3; atrial-specific KO with clonal analysis, transcriptomics and forced Notch activation; ChIP and reporter mutagenesis for Lrrc10

    PMID:23751912 PMID:24563458 PMID:25053429

    Open questions at the time
    • Mechanism linking NKX2-5 loss to Notch derepression unresolved
    • Direct vs. indirect regulation of Rspo3 enhancer not fully mapped
  12. 2015 High

    Revealed a post-transcriptional layer of NKX2-5 control by the RHAU/DHX36 helicase acting on 5'-UTR G-quadruplex (translation) and 3'-UTR AU-rich element (decay).

    Evidence Cardiac-specific Rhau KO mouse, RNA-protein binding, G-quadruplex detection, and ribosomal assays

    PMID:26489465

    Open questions at the time
    • Whether RHAU regulation is dynamically controlled during development unknown
    • Other RNA-binding regulators not surveyed
  13. 2016 Medium

    Identified SIRT1-mediated deacetylation at K182 as a negative regulatory input that weakens NKX2-5 cofactor binding, adding a metabolic/NAD-linked control point.

    Evidence Co-IP, K182 mutagenesis, in vitro deacetylation, and cofactor-binding reporter assays

    PMID:27819261

    Open questions at the time
    • In vivo physiological role of K182 deacetylation not established (single-lab biochemistry)
    • Acetyltransferase counterpart not identified
  14. 2018 High

    Defined human-cell-specific NKX2-5 downstream circuitry, identifying HEY2 as a key mediator of cardiomyogenic differentiation and electrophysiological maturation.

    Evidence NKX2-5 knockout hESC-derived cardiomyocytes with profiling, HEY2 genetic rescue, and electrophysiology

    PMID:29636455

    Open questions at the time
    • Whether HEY2 is a direct NKX2-5 target not established
    • Mechanism of abnormal action potentials not fully resolved
  15. 2022 High

    Established NKX2-5 as a recruiter of the CHD4/NuRD repressor to silence noncardiac programs, and as a requirement for cardiac regeneration, completing a picture of both gene activation and active repression.

    Evidence MS Co-IP, ChIP-seq, and in vivo silencer-deletion mice; zebrafish nkx2-5 KO regeneration assay with RNA-seq

    PMID:35450884 PMID:35624100

    Open questions at the time
    • How NKX2-5 selectively directs CHD4 to silencer vs. activator loci unresolved
    • Regeneration findings are zebrafish-based (Medium confidence) and not validated in mammals
  16. 2019 Medium

    Linked common NKX2-5 regulatory binding variation to human EKG-trait variation, connecting NKX2-5 occupancy to population-level cardiac physiology.

    Evidence Allele-specific ChIP-seq in iPSC-cardiomyocytes with GWAS fine-mapping and reporter validation of two variants

    PMID:31570892

    Open questions at the time
    • Only two of ~2,000 ASE-SNVs experimentally validated
    • Causal target genes for most variants not established

Open questions

Synthesis pass · forward-looking unresolved questions
  • The integration of NKX2-5's multiple regulatory inputs (SUMOylation, deacetylation, RHAU translational control) with its context-specific selection between activation and CHD4-mediated repression at individual loci remains unresolved.
  • No model explains how a single factor switches between activator and repressor at specific targets
  • In vivo physiological roles of individual PTMs largely untested
  • Structure of NKX2-5 in cooperative cofactor complexes undetermined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140110 transcription regulator activity 7 GO:0003677 DNA binding 3
Localization
GO:0005634 nucleus 2
Pathway
R-HSA-74160 Gene expression (Transcription) 5 R-HSA-1266738 Developmental Biology 4 R-HSA-162582 Signal Transduction 3
Partners
CALCHD4GATA4MEF2CSIRT1SRFTBX5
Complex memberships
CHD4/NuRD repressor complex

Evidence

Reading pass · 39 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1997 NKX2-5 and GATA-4 physically interact and are mutual cofactors: the C-terminal zinc finger of GATA-4 and a C-terminus extension interact with the C-terminally extended homeodomain of NKX2-5, resulting in synergistic activation of the ANF promoter. Binding of GATA-4 to the C-terminus autorepressive domain of NKX2-5 may induce a conformational change that unmasks NKX2-5 activation domains. GATA-6 cannot substitute for GATA-4 in this interaction. In vitro binding assays, co-immunoprecipitation, transcription reporter assays (ANF promoter) in heterologous cells, structure-function deletion mapping The EMBO journal High 9312027
1999 Csx/Nkx2-5 acts genetically upstream of multiple cardiac transcription factors and structural genes: in Nkx2-5 null mice, expression of ANF, BNP, MLC2V, N-myc, MEF2C, HAND1, and Msx2 is disturbed. Cardiac development arrests after looping without enhanced apoptosis or reduced proliferation, and Nkx2-5-deficient cells exert dominant interfering effects on cardiac development in chimeras. Csx/Nkx2-5 null mouse knockout, in situ hybridization for 20 candidate genes, TUNEL and PCNA staining, chimeric mouse analysis with ES cells Development (Cambridge, England) High 10021345
2001 NKX2-5 physically associates with TBX5 via the NKX2-5 homeodomain and the N-terminal domain plus N-terminal T-box of TBX5, and this interaction mediates synergistic transcriptional activation of the Nppa (ANF) promoter. The Holt-Oram syndrome TBX5-G80R mutation (causing cardiac defects) abolishes this synergy, whereas R237Q (causing limb defects) retains it. Yeast two-hybrid screen, co-immunoprecipitation in COS-7 cells, GST pull-down assay, promoter-reporter co-transfection, P19CL6 differentiation assay Nature genetics High 11431700
2000 Human CSX/NKX2-5 homeodomain mutations associated with congenital heart disease cause loss of DNA binding and transcriptional activation; homeodomain missense mutations that preserve homodimerization can act in a dominant-negative manner on wild-type CSX/NKX2-5 activity through the ANF promoter. Nuclear localization assay, EMSA (DNA binding), luciferase reporter transcription assays, homodimerization assays with multiple patient-derived mutants The Journal of clinical investigation High 10903346
2000 The NKX2-5 homeodomain mediates homodimerization, with Lys193 and Arg194 at the C-terminal end of the homeodomain being essential for dimerization. Lys193 is also required for specific interaction with GATA4. NKX2-5 can heterodimerize with Nkx2.3 and Nkx2.6 with different affinities. The I183P mutant abolishes DNA binding while preserving homodimerization, and acts as a dominant inhibitor in fibroblasts but not in cardiomyocytes. In vitro protein interaction assays, co-immunoprecipitation, EMSA with monomeric and dimeric binding sites, site-directed mutagenesis, transcription reporter assays The Journal of biological chemistry High 11042197
2007 Nkx2-5 and Tbx5 cooperatively regulate the Id2 promoter in the ventricular conduction system. A 1.2 kb Id2 promoter fragment is sufficient for cooperative regulation by Nkx2-5 and Tbx5 in vitro and for conduction-system-specific expression in vivo. Compound haploinsufficiency of Tbx5 and Nkx2-5 prevents embryonic specification of the ventricular conduction system, placing them in a molecular pathway: Tbx5 → Nkx2-5 → Id2 → conduction system specification. SAGE transcriptional profiling of microdissected conduction system, Id2 knockout mouse analysis, in vitro promoter reporter assays, compound haploinsufficiency genetic epistasis in mice Cell High 17604724
2004 Nkx2-5 gene dosage directly determines the number of cells in the cardiac conduction system: null mutant embryos appear to lack the AV node primordium, and heterozygous Nkx2-5 KO mice have half the normal number of conduction system cells. A connexin40-negative/connexin45-positive cell population is specifically absent from the AV node in heterozygotes. Cellular expression of connexin40 in Purkinje fibers is unaffected. Nkx2-5 knockout and heterozygous mouse analysis, cell counting in conduction system, immunohistochemistry for connexin40 and connexin45, in vivo electrophysiology The Journal of clinical investigation High 15085192
2001 Nkx2-5 is required for eHAND expression in the precardiac mesoderm. In Nkx2-5/dHAND double-null mice, complete ventricular dysgenesis occurs (only atrial chamber forms), and Irx4 expression is cooperatively regulated by Nkx2-5 and dHAND, establishing a genetic pathway for ventricular formation. Double knockout mouse epistasis analysis, molecular marker analysis (eHAND, Irx4), histology, cell fate analysis Developmental biology High 11784028
2001 Expression of a DNA-binding defective Nkx2-5 mutant (I183P) under the beta-MHC promoter in transgenic mice leads to progressive AV conduction block and heart failure, with dramatic decrease in connexin40 and connexin43 expression in transgenic hearts, indicating Nkx2-5 regulates these gap junction genes. Transgenic mouse overexpression of dominant-negative Nkx2-5(I183P), in vivo ECG, connexin immunohistochemistry The Journal of clinical investigation High 11457872
2002 Nkx2-5 is required for cardiomyocyte survival and homeostasis in adult heart: dominant-negative Nkx2-5 transgenic mice show cardiac myocyte degeneration and impaired function; wild-type Nkx2-5 overexpression protects cardiomyocytes from doxorubicin-induced apoptosis; dominant-negative Nkx2-5 induces apoptosis in cultured cardiomyocytes. Transgenic mouse overexpression (dominant-negative and wild-type), doxorubicin treatment, TUNEL apoptosis assay, cardiac function measurements The Journal of biological chemistry High 11889119
2002 Synergistic activation of the cardiac alpha-actin (alphaCA) promoter requires combinatorial binding of SRF, Nkx2-5, and GATA4. DNA-binding defective Nkx2-5pm can coactivate the alphaCA promoter via cooperative SRF binding, and SRF physically associates with Nkx2-5 and GATA4, forming higher-order promoter complexes. Transcription reporter assays in CV1 and Schneider 2 cells, mouse ES cell endogenous gene activation, EMSA, co-transfection with DNA-binding mutants The Journal of biological chemistry High 11983708
2003 Nkx2-5 transcriptionally activates the myocardin promoter. Myocardin expression is markedly downregulated in Nkx2-5-null mouse hearts (identified by subtractive hybridization), and inhibition of myocardin function prevents cardiomyocyte differentiation in P19CL6 cells. Subtractive hybridization from Nkx2-5 null vs. wild-type E8.5 hearts, cotransfection/reporter assay, siRNA inhibition of myocardin in P19CL6 cells Molecular and cellular biology High 14645532
2004 NKX2-5 interacts with a novel LIM domain protein Cal (CSX-associated LIM protein): the LIM domains of Cal and the homeodomain of NKX2-5 are necessary for binding. Cal enhances NKX2-5-induced ANP promoter activation and shuttles from cytoplasm to nucleus in response to calcium to promote cardiomyocyte differentiation. Yeast two-hybrid screening, in vivo and in vitro co-immunoprecipitation, transcription reporter assay, subcellular localization by microscopy, P19CL6 differentiation assay The Journal of cell biology High 14757752
2008 NKX2-5 is SUMOylated at Lys51 (K51), a conserved residue absent in other NK-2 family members. This SUMOylation is catalyzed by PIAS1, PIASx, and PIASy (but not PIAS3) for SUMO-1, and only by PIASx for SUMO-2. SUMO modification stabilizes NKX2-5-containing complexes and robustly enhances transcriptional activity; mutation K51R reduces DNA binding and transcriptional activity, and the mutant is targeted by ubiquitin instead. In vitro SUMOylation assays, site-directed mutagenesis (K51R), E3 ligase co-expression assays, transcription reporter assays The Journal of biological chemistry High 18579533
2011 SUMOylation of NKX2-5 strongly enhances transcriptional activity; K51 is a SUMOylation target but is not the only site — at least one additional, non-canonical or shifting SUMOylation site exists that is the predominant site in cardiac cells and cannot be abrogated by mutation of any individual lysine in the K51R background. Biochemical SUMOylation assays, systematic mutagenesis of all lysines in K51R background, transcription reporter assays across multiple cell lines PloS one Medium 21931855
2016 SIRT1 deacetylates NKX2-5 at lysine 182 within the homeodomain. SIRT1 interacts with the C-terminus of NKX2-5. Mutation of K182 reduces NKX2-5 transcriptional activity. SIRT1 inhibits NKX2-5 transcriptional activity partly by reducing NKX2-5 binding to its cofactors SRF and TBX5. Co-immunoprecipitation, site-directed mutagenesis (K182), in vitro deacetylation assays, transcription reporter assays, co-factor binding assays Scientific reports Medium 27819261
2008 Direct physical interaction between NKX2-5 and MEF2C was demonstrated. Compound Nkx2-5(-/-);Mef2c(-/-) double mutants lack identifiable ventricles (single mutants both have ventricles), with mutant cardiomyocytes expressing only atrial/second heart field markers, indicating cooperative requirement for ventricular differentiation. Co-immunoprecipitation, mammalian two-hybrid assay, genetic double-knockout epistasis, histological and molecular phenotyping Developmental dynamics High 19035347
2007 Nkx2-5 and its target Cx40 are expressed in atria and pulmonary myocardium but not in systemic venous return. When Nkx2-5 protein level is lowered in a hypomorphic model, pulmonary myocardium switches to a Cx40-negative, Hcn4-positive phenotype resembling systemic venous return, establishing Nkx2-5 as a determinant of pulmonary myocardial identity. Genetic labeling shows pulmonary myocardium derives from Nkx2-5-expressing precursors, distinct from systemic venous Nkx2-5-negative precursors. Nkx2-5 hypomorphic mouse model, genetic Cre-lox lineage labeling, immunohistochemistry for Cx40 and Hcn4 Circulation research High 17823370
2008 Perinatal loss of Nkx2-5 (tamoxifen-inducible KO) results in reduction of cardiac Nav1.5-alpha (voltage-gated Na+ channel pore-forming subunit) and ryanodine receptor 2 expression, accompanied by conduction and contraction defects within 4 days, demonstrating that Nkx2-5 regulates these ion channel/Ca2+ handling genes postnatally. Tamoxifen-inducible conditional Nkx2-5 KO mouse, ECG, RT-PCR/Western blot for Nav1.5 and RyR2 Circulation research High 18689573
2011 Ectopic overexpression of Nkx2-5 in the developing mouse heart leads to severe hypoplasia of the sinoatrial node (SAN) and venous valves, converting SAN cell fate toward working myocardium, and causes embryonic lethality. Shox2 normally suppresses Nkx2-5 in the SAN, and Shox2 acts upstream of Nkx2-5 to shield SAN from becoming working myocardium. Tissue-specific transgenic overexpression of Nkx2-5 in mouse heart, in situ hybridization and immunohistochemistry for SAN markers, ECG Developmental biology High 21640717
2011 NKX2-5 directly represses Isl1 transcription: NKX2-5 binds to an Isl1 enhancer and represses Isl1 transcriptional activity. Nkx2-5-deficient embryos fail to downregulate Isl1 protein in heart tube cardiomyocytes. This Isl1/Nkx2-5 mechanism coordinates specification of cardiac progenitors toward ventricular vs. nodal myocyte identities. Mouse ESC overexpression/KO, Xenopus gain-of-function, chromatin binding assay (Nkx2-5 binding to Isl1 enhancer), luciferase reporter, single-cell electrophysiology, immunocytochemistry Stem cells (Dayton, Ohio) High 25524439
2012 Crystal structure of the NKX2-5 homeodomain in complex with double-stranded DNA at 1.8 Å resolution reveals that two homeodomains bind both sites of the ANF proximal promoter (-242 site) separated by five nucleotides without physical interaction between themselves, with identical conformations at both sites. Tyr54, conserved in all NK2 proteins, mediates sequence-specific interaction with the TAAG motif. X-ray crystallography (1.8 Å resolution), structural analysis of protein-DNA contacts Biochemistry High 22849347
2014 Nkx2-5 regulates cardiac growth in the second heart field (SHF) by transcriptionally activating R-spondin3 (Rspo3), a secreted Wnt agonist. Rspo3 is markedly downregulated in Nkx2-5 mutants; conditional Rspo3 inactivation in the Isl1 lineage phenocopies Nkx2-5 SHF defects; enhancing Wnt/β-catenin signaling pharmacologically or by transgenic Rspo3 rescues SHF defects in Nkx2-5 conditional heterozygous mutants. Nkx2-5 conditional KO, Rspo3 conditional KO, pharmacological Wnt activation, Rspo3 transgenic rescue, gene expression analysis Development (Cambridge, England) High 25053429
2014 Nkx2-5 suppresses cardiomyocyte proliferation: atrial-specific deletion of Nkx2-5 results in hyperplastic working myocytes and conduction system. Transcriptome analysis reveals aberrant Notch signaling activation underlies the hyperproliferation, and forced Notch activation recapitulates hyperproliferation of working myocytes. Atrial-specific Nkx2-5 conditional KO mouse, multicolor reporter clonal analysis, transcriptome profiling, forced Notch pathway activation Circulation research High 24563458
2015 RHAU (DHX36) RNA helicase post-transcriptionally regulates Nkx2-5: RHAU associates with the 5' UTR and 3' UTR of Nkx2-5 mRNA. The 5' UTR contains a G-quadruplex requiring RHAU for protein translation, while the 3' UTR AU-rich element facilitates RHAU-mediated mRNA decay. Cardiac Rhau deletion causes heart defects and embryonic lethality with reduced Nkx2-5 expression. Cardiac-specific Rhau KO mouse, gene expression profiling, RNA-protein binding assays, G-quadruplex detection, ribosomal assays Cell reports High 26489465
2011 NKX2-5 regulates Cdc42 indirectly via miR-1: Nkx2-5 negatively regulates miR-1 expression in the mouse heart (and Tinman does so in Drosophila), and miR-1 negatively regulates Cdc42. Compound Nkx2-5/Cdc42 heterozygous mutant mice show conduction system and cardiac output defects, demonstrating a conserved Tinman/Nkx2-5 → miR-1 → Cdc42 pathway regulating heart function. Compound heterozygous mouse genetics, miR-1 expression analysis, Cdc42 targeting validation, Drosophila genetic interaction, cardiac output measurements The Journal of cell biology High 21690310
2018 NKX2-5 regulates cardiomyogenesis through a HEY2-dependent transcriptional network: NKX2-5 null hESC-derived cardiomyocytes fail to activate VCAM1, fail to downregulate PDGFRα, have abnormal action potentials, and HEY2 is identified as a key mediator of NKX2-5 function by molecular profiling and genetic rescue experiments. NKX2-5 knockout hESCs, molecular profiling, genetic rescue (HEY2 restoration), electrophysiology, flow cytometry for surface markers Nature communications High 29636455
2022 CHD4 (catalytic subunit of NuRD complex) interacts with NKX2-5, GATA4, and TBX5 during embryonic heart development. The NKX2-5-CHD4 complex occupies specific cardiac loci to repress noncardiac gene programs. Deletion of NKX2-5-CHD4-controlled silencer elements for Acta1 and Myh11 leads to inappropriate skeletal and smooth muscle gene expression in embryonic heart. Mass spectrometry co-immunoprecipitation, ChIP-seq occupancy mapping, transcriptomics, in vivo silencer deletion mouse models Genes & development High 35450884
2019 NKX2-5 allele-specific binding at ~2,000 common regulatory variants (ASE-SNVs) in iPSC-derived cardiomyocytes contributes to EKG trait variation. Two NKX2-5 ASE-SNVs (rs3807989 and rs590041) modulate target gene expression via differential protein binding in cardiac cells. iPSC-derived cardiomyocytes from related individuals, allele-specific ChIP-seq, GWAS fine-mapping, reporter assays for two validated variants Nature genetics Medium 31570892
2013 Lrrc10 is a direct transcriptional target of Nkx2-5 and GATA4: Nkx2-5 and GATA4 endogenously occupy the proximal and distal cardiac regulatory elements of the Lrrc10 promoter. Nkx2-5 KO mouse hearts have dramatically reduced Lrrc10 expression. The proximal element is synergistically activated by Nkx2-5 and GATA4; the distal element requires SRF in addition. ChIP assay (endogenous occupancy), Nkx2-5 KO expression analysis, transcription reporter assay, mutational analysis of binding sites Journal of molecular and cellular cardiology High 23751912
2009 NKX2-5 suppresses beta-catenin expression and activates GATA4 expression in human fetal cardiac myocytes by binding to specific NKX2-5 binding elements (NKEs) in the promoters of these genes; mutational analysis of the NKEs demonstrates they are essential for these regulatory effects. Antisense inhibition of NKX2-5, promoter-luciferase reporter with NKE mutagenesis, gene expression analysis PloS one Medium 19479054
2017 NKX2-5 point mutations disrupt the Wnt signaling pathway in cardiomyocytes, promoting heart dysfunction through alteration of cardiomyocyte metabolism. A murine Nkx2-5 point mutation model recapitulates the full spectrum of human congenital heart disease phenotypes including right ventricular dysfunction. Murine Nkx2-5 point mutation knock-in model, transcriptional network analysis, Wnt pathway assays, metabolic profiling JCI insight Medium 28352650
2022 Nkx2-5 is required for cardiac regeneration in zebrafish: nkx2-5-null fish have impaired ventricular apex regeneration with diminished dedifferentiation and proliferation. Nkx2-5 activates proteolytic pathways for sarcomere disassembly and promotes cardiomyocyte proliferative response; RNA-seq reveals disrupted embryonic transcriptional profile in adult Nkx2-5 loss-of-function myocardium. Nkx2-5 KO zebrafish, ventricular apex amputation, cardiac-specific RNA-seq, gene regulatory network analysis Nature communications Medium 35624100
2006 Nkx2-5 is cell-autonomously required for postnatal differentiation of Purkinje fibers: chimeric analysis shows maximal Nkx2-5 levels are required in a cell-autonomous manner; deficiency in Purkinje fibers becomes measurable only after birth. Reduced Nkx2-5 is associated with delayed cell cycle withdrawal in surrounding myocytes. Nkx2-5+/- × Cx40(eGFP/+) cross for conduction system visualization, chimeric embryo analysis, electrophysiology, cell cycle analysis Developmental biology High 17250822
2019 Nkx2-5 defines a subpopulation of pacemaker cells in the SAN transitional junction zone. Although Nkx2-5 is dispensable for SAN morphogenesis during embryogenesis, its deletion in SAN junction cells hampers atrial activation by the pacemaker, demonstrating a functional role in pacemaker exit conduction. Conditional Nkx2-5 KO in SAN junction, single-cell RNA-seq, action potential recording, ECG Development (Cambridge, England) High 31320323
1999 Hmx1 and NKX2-5 bind to the same DNA consensus sequence (5'-CAAGTG-3') with high affinity; co-expression of Hmx1 and Nkx2-5 results in transcriptional antagonism — NKX2-5 transactivates reporters containing this element while Hmx1 represses them. SAAB DNA binding site selection, EMSA, luciferase reporter assays with co-transfection, mutagenesis of binding site The Journal of biological chemistry Medium 10206974
2007 Csx/Nkx2-5 overexpression in neonatal rat cardiomyocytes markedly increases spontaneous beating rate by upregulating the Cav3.2 T-type Ca2+ channel, while downregulating the L-type Ca2+ channel (Cav1.3 mRNA dramatically decreased). These are distinct from GATA4 effects. Adenoviral overexpression in neonatal rat cardiomyocytes, patch clamp electrophysiology, real-time PCR Journal of molecular and cellular cardiology Medium 17498735
2011 Conditional Nkx2-5 ablation beginning at E12.5 results in embryonic death by E17.5 with ASD, arrhythmias, and contraction defects. Within 4 days of Nkx2-5 ablation, expression of Nav1.5-alpha, connexin40, cardiac myosin light chain kinase, and sarcolipin is abnormal, identifying these as downstream targets. Tamoxifen-inducible conditional KO beginning at E12.5, 3D serial section reconstruction, quantitative RT-PCR, ECG Cardiovascular research High 21285290
2011 NKX2-5 is expressed in Nkx2-5+ cardiac progenitors that contribute to the proepicardium (which expresses Wt1 and Tbx18). Nkx2-5 knockout results in abnormal proepicardial development and decreased Wt1 expression, demonstrating a functional role for Nkx2-5 in proepicardium formation. In vivo Cre-loxP lineage tracing using Nkx2-5-Cre, immunofluorescence, Nkx2-5 KO analysis Biochemical and biophysical research communications Medium 18722343

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1997 The cardiac transcription factors Nkx2-5 and GATA-4 are mutual cofactors. The EMBO journal 531 9312027
1999 The cardiac homeobox gene Csx/Nkx2.5 lies genetically upstream of multiple genes essential for heart development. Development (Cambridge, England) 468 10021345
1993 Csx: a murine homeobox-containing gene specifically expressed in the developing heart. Proceedings of the National Academy of Sciences of the United States of America 456 7690144
2001 Tbx5 associates with Nkx2-5 and synergistically promotes cardiomyocyte differentiation. Nature genetics 455 11431700
2011 NKX2-5(eGFP/w) hESCs for isolation of human cardiac progenitors and cardiomyocytes. Nature methods 326 22020065
2007 Pitx2c and Nkx2-5 are required for the formation and identity of the pulmonary myocardium. Circulation research 299 17823370
2001 NKX2.5 mutations in patients with tetralogy of fallot. Circulation 259 11714651
2001 Embryonic expression of an Nkx2-5/Cre gene using ROSA26 reporter mice. Genesis (New York, N.Y. : 2000) 258 11783008
2007 A molecular pathway including Id2, Tbx5, and Nkx2-5 required for cardiac conduction system development. Cell 223 17604724
2004 Nkx2-5 mutation causes anatomic hypoplasia of the cardiac conduction system. The Journal of clinical investigation 192 15085192
2005 Cardiac transcription factor Csx/Nkx2-5: Its role in cardiac development and diseases. Pharmacology & therapeutics 174 15925411
2005 Phenotypes with GATA4 or NKX2.5 mutations in familial atrial septal defect. American journal of medical genetics. Part A 164 15810002
1999 Building the heart piece by piece: modularity of cis-elements regulating Nkx2-5 transcription. Development (Cambridge, England) 142 10477287
2001 The combinatorial activities of Nkx2.5 and dHAND are essential for cardiac ventricle formation. Developmental biology 136 11784028
2000 Loss of function and inhibitory effects of human CSX/NKX2.5 homeoprotein mutations associated with congenital heart disease. The Journal of clinical investigation 134 10903346
1998 Cardiac and extracardiac expression of Csx/Nkx2.5 homeodomain protein. Circulation research 129 9598591
2002 Combinatorial expression of GATA4, Nkx2-5, and serum response factor directs early cardiac gene activity. The Journal of biological chemistry 128 11983708
2001 Progressive atrioventricular conduction defects and heart failure in mice expressing a mutant Csx/Nkx2.5 homeoprotein. The Journal of clinical investigation 120 11457872
2008 Nkx2-5- and Isl1-expressing cardiac progenitors contribute to proepicardium. Biochemical and biophysical research communications 116 18722343
1999 Context-dependent transcriptional cooperation mediated by cardiac transcription factors Csx/Nkx-2.5 and GATA-4. The Journal of biological chemistry 102 10075728
1999 Identification of upstream regulatory regions in the heart-expressed homeobox gene Nkx2-5. Development (Cambridge, England) 98 9895330
2003 Myocardin expression is regulated by Nkx2.5, and its function is required for cardiomyogenesis. Molecular and cellular biology 93 14645532
2010 NKX2-5: an update on this hypermutable homeodomain protein and its role in human congenital heart disease (CHD). Human mutation 92 20725931
2018 NKX2-5 regulates human cardiomyogenesis via a HEY2 dependent transcriptional network. Nature communications 89 29636455
2010 Mutational spectrum in the cardiac transcription factor gene NKX2.5 (CSX) associated with congenital heart disease. Clinical genetics 83 20456451
2008 Perinatal loss of Nkx2-5 results in rapid conduction and contraction defects. Circulation research 82 18689573
2000 Characterization of homo- and heterodimerization of cardiac Csx/Nkx2.5 homeoprotein. The Journal of biological chemistry 81 11042197
2008 Cooperative interaction of Nkx2.5 and Mef2c transcription factors during heart development. Developmental dynamics : an official publication of the American Association of Anatomists 79 19035347
2008 WNT signaling promotes Nkx2.5 expression and early cardiomyogenesis via downregulation of Hdac1. Biochimica et biophysica acta 78 18851995
2015 Familial Atrial Septal Defect and Sudden Cardiac Death: Identification of a Novel NKX2-5 Mutation and a Review of the Literature. Congenital heart disease 76 26679770
2001 Nkx2-5 activity is essential for cardiomyogenesis. The Journal of biological chemistry 76 11526122
2016 Genetics of Congenital Heart Defects: The NKX2-5 Gene, a Key Player. Genes 73 26805889
2019 Structure of Csx1-cOA4 complex reveals the basis of RNA decay in Type III-B CRISPR-Cas. Nature communications 71 31541109
2004 Novel NKX2-5 mutations in diseased heart tissues of patients with cardiac malformations. The American journal of pathology 71 15161646
1999 Complex modular cis-acting elements regulate expression of the cardiac specifying homeobox gene Csx/Nkx2.5. Development (Cambridge, England) 71 10068637
2015 Direct nkx2-5 transcriptional repression of isl1 controls cardiomyocyte subtype identity. Stem cells (Dayton, Ohio) 69 25524439
2006 Wnt signaling and CEH-22/tinman/Nkx2.5 specify a stem cell niche in C. elegans. Current biology : CB 69 16461282
2015 The CRISPR-associated Csx1 protein of Pyrococcus furiosus is an adenosine-specific endoribonuclease. RNA (New York, N.Y.) 68 26647461
2011 Tinman/Nkx2-5 acts via miR-1 and upstream of Cdc42 to regulate heart function across species. The Journal of cell biology 68 21690310
2007 RNA toxicity in myotonic muscular dystrophy induces NKX2-5 expression. Nature genetics 68 18084293
2002 Novel point mutation in the cardiac transcription factor CSX/NKX2.5 associated with congenital heart disease. Circulation journal : official journal of the Japanese Circulation Society 68 12074273
2011 Ablation of Nkx2-5 at mid-embryonic stage results in premature lethality and cardiac malformation. Cardiovascular research 66 21285290
1996 Molecular cloning and characterization of human cardiac homeobox gene CSX1. Circulation research 66 8888684
2015 Post-transcriptional Regulation of Nkx2-5 by RHAU in Heart Development. Cell reports 65 26489465
2014 A novel NKX2.5 loss-of-function mutation associated with congenital bicuspid aortic valve. The American journal of cardiology 64 25438918
2006 A novel CSX/NKX2-5 mutation causes autosomal-dominant AV block: are atrial fibrillation and syncopes part of the phenotype? European journal of human genetics : EJHG 64 16896344
2006 Nkx2.5 cell-autonomous gene function is required for the postnatal formation of the peripheral ventricular conduction system. Developmental biology 64 17250822
2011 Ectopic expression of Nkx2.5 suppresses the formation of the sinoatrial node in mice. Developmental biology 63 21640717
2002 Csx/Nkx2-5 is required for homeostasis and survival of cardiac myocytes in the adult heart. The Journal of biological chemistry 61 11889119
2014 Nkx2-5 regulates cardiac growth through modulation of Wnt signaling by R-spondin3. Development (Cambridge, England) 58 25053429
2000 Subdivision of the cardiac Nkx2.5 expression domain into myogenic and nonmyogenic compartments. Developmental biology 58 10656773
2014 Mesodermal Nkx2.5 is necessary and sufficient for early second heart field development. Developmental biology 57 24613616
2013 A novel NKX2.5 loss-of-function mutation responsible for familial atrial fibrillation. International journal of molecular medicine 52 23525379
2004 A novel LIM protein Cal promotes cardiac differentiation by association with CSX/NKX2-5. The Journal of cell biology 52 14757752
2014 A novel NKX2-5 loss-of-function mutation predisposes to familial dilated cardiomyopathy and arrhythmias. International journal of molecular medicine 51 25503402
2014 Nkx2-5 suppresses the proliferation of atrial myocytes and conduction system. Circulation research 50 24563458
2000 Functional analyses of three Csx/Nkx-2.5 mutations that cause human congenital heart disease. The Journal of biological chemistry 50 10948187
2013 DNA methylation status of NKX2-5, GATA4 and HAND1 in patients with tetralogy of fallot. BMC medical genomics 49 24182332
2008 NKX2.5 mutations in patients with non-syndromic congenital heart disease. International journal of cardiology 49 19073351
2022 Computational profiling of hiPSC-derived heart organoids reveals chamber defects associated with NKX2-5 deficiency. Communications biology 47 35488063
2017 Discovery of Small Molecules Targeting the Synergy of Cardiac Transcription Factors GATA4 and NKX2-5. Journal of medicinal chemistry 47 28858485
2008 Regulation of cardiac specific nkx2.5 gene activity by small ubiquitin-like modifier. The Journal of biological chemistry 45 18579533
2009 Nkx2.7 and Nkx2.5 function redundantly and are required for cardiac morphogenesis of zebrafish embryos. PloS one 44 19158954
2007 Transcription factors Csx/Nkx2.5 and GATA4 distinctly regulate expression of Ca2+ channels in neonatal rat heart. Journal of molecular and cellular cardiology 44 17498735
1998 Upregulation of the cardiac homeobox gene Nkx2-5 (CSX) in feline right ventricular pressure overload. The American journal of physiology 43 9612365
2019 Allele-specific NKX2-5 binding underlies multiple genetic associations with human electrocardiographic traits. Nature genetics 42 31570892
2009 NKX2-5 regulates the expression of beta-catenin and GATA4 in ventricular myocytes. PloS one 41 19479054
1999 Transcriptional antagonism between Hmx1 and Nkx2.5 for a shared DNA-binding site. The Journal of biological chemistry 41 10206974
2012 Crystal structure of the human NKX2.5 homeodomain in complex with DNA target. Biochemistry 37 22849347
2022 CHD4 is recruited by GATA4 and NKX2-5 to repress noncardiac gene programs in the developing heart. Genes & development 36 35450884
2003 Early stage-specific inhibitions of cardiomyocyte differentiation and expression of Csx/Nkx-2.5 and GATA-4 by phosphatidylinositol 3-kinase inhibitor LY294002. Experimental cell research 36 14597408
2016 Mutations of NKX2.5 and GATA4 genes in the development of congenital heart disease. Gene 34 27154817
2011 Complex SUMO-1 regulation of cardiac transcription factor Nkx2-5. PloS one 34 21931855
2018 Cardiac Actions of a Small Molecule Inhibitor Targeting GATA4-NKX2-5 Interaction. Scientific reports 33 29545582
2014 Mutational spectrum of the NKX2-5 gene in patients with lone atrial fibrillation. International journal of medical sciences 33 24782644
2012 Crystal structure and nucleic acid-binding activity of the CRISPR-associated protein Csx1 of Pyrococcus furiosus. Proteins 33 22987782
2011 Investigation of somatic NKX2-5, GATA4 and HAND1 mutations in patients with tetralogy of Fallot. Pathology 33 21519287
2005 CSX/Nkx2.5 modulates differentiation of skeletal myoblasts and promotes differentiation into neuronal cells in vitro. The Journal of biological chemistry 33 15653675
2017 Allosteric regulation of Csx1, a type IIIB-associated CARF domain ribonuclease by RNAs carrying a tetraadenylate tail. Nucleic acids research 32 28977519
2015 NKX2-5 mutations in an inbred consanguineous population: genetic and phenotypic diversity. Scientific reports 32 25742962
1995 Assignment of cardiac homeobox gene CSX to human chromosome 5q34. Genomics 32 7665173
2017 Nkx2.5 marks angioblasts that contribute to hemogenic endothelium of the endocardium and dorsal aorta. eLife 30 28271994
2006 Differentiation of cardiac Purkinje fibers requires precise spatiotemporal regulation of Nkx2-5 expression. Developmental dynamics : an official publication of the American Association of Anatomists 30 16245335
2016 SIRT1 deacetylates the cardiac transcription factor Nkx2.5 and inhibits its transcriptional activity. Scientific reports 28 27819261
2010 A novel NKX2-5 mutation in familial ventricular septal defect. International journal of molecular medicine 28 21165553
2017 Prevalence and spectrum of NKX2.5 mutations in patients with congenital atrial septal defect and atrioventricular block. Molecular medicine reports 27 28259982
2005 Sox9 and Nkx2.5 determine the pyloric sphincter epithelium under the control of BMP signaling. Developmental biology 27 15733673
2019 Nkx2-5 defines a subpopulation of pacemaker cells and is essential for the physiological function of the sinoatrial node in mice. Development (Cambridge, England) 26 31320323
2013 Nkx2-5 mediates differential cardiac differentiation through interaction with Hoxa10. Stem cells and development 26 23477547
2013 Lrrc10 is a novel cardiac-specific target gene of Nkx2-5 and GATA4. Journal of molecular and cellular cardiology 26 23751912
2020 Nkx2-5 defines distinct scaffold and recruitment phases during formation of the murine cardiac Purkinje fiber network. Nature communications 25 33082351
2017 Nkx2.5+ Cardiomyoblasts Contribute to Cardiomyogenesis in the Neonatal Heart. Scientific reports 25 28974782
2011 Mutations in the NKX2.5 gene and the PAX8 promoter in a girl with thyroid dysgenesis. The Journal of clinical endocrinology and metabolism 25 21450989
2022 Activation of Nkx2.5 transcriptional program is required for adult myocardial repair. Nature communications 24 35624100
2006 Single-cell-derived mesenchymal stem cells overexpressing Csx/Nkx2.5 and GATA4 undergo the stochastic cardiomyogenic fate and behave like transient amplifying cells. Experimental cell research 24 17208226
2014 Pharmacological inhibition of TGFβ receptor improves Nkx2.5 cardiomyoblast-mediated regeneration. Cardiovascular research 23 25362681
2011 Overexpression of Csx/Nkx2.5 and GATA-4 enhances the efficacy of mesenchymal stem cell transplantation after myocardial infarction. Circulation journal : official journal of the Japanese Circulation Society 23 21828931
2017 Point mutations in murine Nkx2-5 phenocopy human congenital heart disease and induce pathogenic Wnt signaling. JCI insight 22 28352650
2010 Novel NKX2-5 mutations in patients with familial atrial septal defects. Pediatric cardiology 22 21188375
2004 Function follows form: cardiac conduction system defects in Nkx2-5 mutation. The anatomical record. Part A, Discoveries in molecular, cellular, and evolutionary biology 22 15368343

Missed literature

Know a paper Affinage missed for NKX2-5? Flag it for the maintainers and the community.

No submissions yet.