Affinage

NEK2

Serine/threonine-protein kinase Nek2 · UniProt P51955

Length
445 aa
Mass
51.8 kDa
Annotated
2026-04-29
100 papers in source corpus 46 papers cited in narrative 46 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

NEK2 is a centrosomal serine/threonine kinase that orchestrates centrosome disjunction, bipolar spindle formation, kinetochore–microtubule attachment, cilium disassembly, and—through phosphorylation of diverse non-mitotic substrates—signal transduction, alternative splicing, and protein stability control. NEK2 forms leucine-zipper-mediated homodimers that undergo trans-autophosphorylation for activation, and its activity is positively regulated by the MAPK/p90Rsk2 and Plk1–Mst2 pathways while negatively regulated by PP1α, kendrin, and HEF1 (PMID:10347187, PMID:11923207, PMID:21723128, PMID:17283141, PMID:16184168). At mitotic entry, NEK2A phosphorylates centrosomal linker components (C-Nap1, Rootletin, β-catenin, Dishevelled, Cep68) to dissolve the intercentrosomal linker, and phosphorylates kinetochore substrates Hec1 and Sgo1 to ensure faithful chromosome segregation, after which NEK2A is destroyed via a unique MR-dipeptide-dependent direct binding to APC/C that bypasses the spindle assembly checkpoint (PMID:18086858, PMID:27486244, PMID:18297113, PMID:17621308, PMID:16648845, PMID:32307883). Beyond mitosis, NEK2 phosphorylates Kif24 to drive cilium resorption, stabilizes EZH2, YAP1, and PD-L1 by blocking their ubiquitin-dependent degradation, inhibits p53 transcriptional activity through Ser315 phosphorylation, activates non-canonical NF-κB signaling via NIK phosphorylation, and modulates alternative splicing through interaction with hnRNPA1/2 (PMID:26290419, PMID:28737508, PMID:35705994, PMID:34315872, PMID:30266789, PMID:35031599, PMID:28086949).

Mechanistic history

Synthesis pass · year-by-year structured walk · 21 steps
  1. 1999 High

    Establishing how NEK2 is activated resolved the fundamental question of how this kinase achieves catalytic competence: leucine-zipper-mediated homodimerization enables trans-autophosphorylation required for full kinase activity.

    Evidence Glycerol gradient sedimentation, yeast two-hybrid, co-precipitation, and in vitro kinase assays with leucine zipper deletion mutants in human cells

    PMID:10347187

    Open questions at the time
    • Structural basis of leucine-zipper-mediated dimerization not yet resolved at atomic level
    • In vivo stoichiometry of dimers versus monomers unknown
  2. 2001 High

    Discovery that NEK2A is destroyed at mitotic entry by APC/C-Cdc20 via D-box and KEN-box motifs explained how the kinase is temporally restricted to the G2/M transition, while isoform NEK2B escapes this regulation.

    Evidence Cell synchronization, proteasome inhibitor treatment, APC/C ubiquitination reconstitution, and destruction motif mutagenesis

    PMID:11742988

    Open questions at the time
    • Mechanism by which NEK2A directly binds APC/C was not yet understood
    • Physiological role of NEK2B persistence in mitosis not defined
  3. 2002 High

    Identification of NEK2 splice variants (Nek2A vs Nek2B) and demonstration that only Nek2A induces centrosome splitting established isoform-specific functional specialization, while localization studies placed NEK2 at centrosomes, chromosomes, and the midbody.

    Evidence RT-PCR, antisense knockdown, overexpression centrosome splitting assay, and immunofluorescence throughout mitosis

    PMID:11742531 PMID:11785960

    Open questions at the time
    • Functional roles at chromosomes and midbody not mechanistically defined
    • Regulatory basis of isoform-specific centrosome splitting unknown
  4. 2002 High

    Placing NEK2 downstream of the MAPK/p90Rsk2 pathway during meiotic G2/M established the first upstream activation mechanism for NEK2 kinase activity.

    Evidence MEK inhibitor treatment during meiotic progression in mouse spermatocytes and in vitro kinase assay with recombinant p90Rsk2

    PMID:11923207

    Open questions at the time
    • Direct p90Rsk2 phosphorylation site(s) on NEK2 not mapped
    • Whether this pathway operates in somatic mitosis not established
  5. 2003 High

    Demonstrating that kinase-dead NEK2A causes monopolar spindles and cytokinesis failure proved that NEK2A catalytic activity is essential for centrosome disjunction and bipolar spindle formation.

    Evidence Tetracycline-inducible overexpression of wild-type vs kinase-dead NEK2A with live cell imaging in human cells

    PMID:12857871

    Open questions at the time
    • Critical centrosomal substrates mediating the splitting phenotype not yet identified
  6. 2004 High

    Identification of NEK2A at kinetochores and demonstration that it recruits MAD2 (but not MAD1) for spindle checkpoint signaling expanded NEK2 function beyond centrosomes to chromosome segregation fidelity.

    Evidence Reciprocal co-IP of NEK2A–MAD1, siRNA knockdown, nocodazole challenge, and immunofluorescence

    PMID:14978040

    Open questions at the time
    • Whether NEK2A phosphorylates MAD1 or MAD2 directly at kinetochores not resolved
    • Mechanism of MAD2 recruitment by NEK2A unclear
  7. 2004 High

    Demonstrating that NEK2A phosphorylates numatrin/NPM and controls its centrosomal localization, and that NEK2A binds Erk2 to recruit it to centrosomes, expanded the roster of NEK2 centrosomal substrates and interactors.

    Evidence Reciprocal co-IP, GST pulldown, in vitro kinase assay, siRNA depletion with localization readout

    PMID:15358203 PMID:15388344

    Open questions at the time
    • Functional consequence of numatrin phosphorylation for centrosome duplication/separation not defined
    • Erk2 interaction confirmed by single co-IP only
  8. 2005 High

    Discovery that NEK2 trafficking to centrosomes requires microtubules, the centriolar satellite component PCM-1, and local proteasomal turnover revealed the dynamic supply chain maintaining centrosomal NEK2 pools.

    Evidence FRAP (t½ ~3 s), microtubule depolymerization, PCM-1 siRNA, motif mutagenesis, proteasome inhibitor treatment

    PMID:15659651

    Open questions at the time
    • Motor protein(s) responsible for NEK2 transport not identified
    • How proteasomal degradation sustains recruitment is mechanistically unclear
  9. 2005 High

    Identification of HEF1 as a negative regulator and Nlp as a phosphorylation-priming substrate of NEK2 at centrosomes revealed multi-layered control of centrosome cohesion and maturation.

    Evidence Co-IP, siRNA depletion causing centrosomal splitting and Nek2 hyperactivation, in vitro kinase assay showing Nek2 phosphorylation primes Nlp for Plk1

    PMID:15684383 PMID:16184168

    Open questions at the time
    • Mechanism by which HEF1 restrains NEK2 activity not defined at the molecular level
  10. 2006 High

    Solving the crystal structure of NEK2's kinase domain revealed an auto-inhibitory helical motif in the activation loop, while discovery of the MR-dipeptide mechanism showed NEK2A bypasses the spindle assembly checkpoint by directly binding APC/C without requiring Cdc20 adaptor function.

    Evidence X-ray crystallography at 2.2 Å, mass spectrometry of autophosphorylation sites, APC/C pull-down with MR-mutation and Cdc20 immunodepletion

    PMID:16648845 PMID:17197699

    Open questions at the time
    • Structural basis of the MR-dipeptide–APC/C interaction not resolved
    • How auto-inhibition is relieved in vivo not fully understood
  11. 2007 High

    Identification of β-catenin, Rootletin, Sgo1, centrobin, and Dishevelled as direct NEK2 substrates, plus PP1α as a DNA-damage-responsive negative regulator, established the core centrosomal linker dissolution mechanism and expanded kinetochore functions.

    Evidence In vitro kinase assays, phospho-specific antibodies, co-IP, electron microscopy, non-phosphorylatable mutant phenotypes, siRNA with centrosome/kinetochore readouts

    PMID:17283141 PMID:17535851 PMID:17621308 PMID:18086858

    Open questions at the time
    • Relative contributions of individual linker substrates to centrosome cohesion not dissected
    • Sgo1 phosphorylation sites' structural impact on cohesin protection not defined
  12. 2008 High

    Demonstrating that NEK2A phosphorylates Hec1 at Ser165 to modulate Ndc80 complex–microtubule binding affinity established a direct mechanism by which NEK2 controls kinetochore–microtubule attachment fidelity.

    Evidence In vitro kinase assay, phospho-specific antibody, non-phosphorylatable mutant causing attachment errors, in vitro microtubule-binding reconstitution

    PMID:18297113

    Open questions at the time
    • Whether Hec1 S165 phosphorylation is the sole NEK2-dependent kinetochore signal is unclear
    • Temporal coordination with Aurora B error-correction pathway not resolved
  13. 2010 High

    Placing Hippo pathway components Mst2 and hSav1 upstream of NEK2A, and subsequently showing that Plk1 phosphorylation of Mst2 relieves PP1γ-mediated suppression of NEK2A, established a Plk1–Mst2–NEK2A signaling axis for centrosome disjunction.

    Evidence Co-IP, siRNA, kinase activity assays, Eg5 inhibitor epistasis, Plk1 inhibition/depletion

    PMID:21076410 PMID:21723128

    Open questions at the time
    • Direct Plk1 phosphorylation sites on Mst2 that control PP1γ binding not mapped
    • Whether this axis operates in all cell types not established
  14. 2015 High

    Discovery that NEK2 phosphorylates Kif24 to stimulate microtubule depolymerization and suppress ciliogenesis, independently of Aurora A/HDAC6, revealed a major non-mitotic function in cilium disassembly.

    Evidence Co-IP, in vitro kinase assay, siRNA knockdown/rescue, ciliation assay in proliferating cells

    PMID:26290419

    Open questions at the time
    • Phosphorylation sites on Kif24 not fully mapped
    • How NEK2–Kif24 axis is coordinated with cell cycle re-entry signals unclear
  15. 2016 High

    Demonstrating that NEK2 phosphorylates Dishevelled to create binding interfaces for CDK5RAP2 and C-NAP1 linked centrosome linker dissolution to Wnt/PCP pathway components, and Drosophila genetic studies confirmed evolutionary conservation.

    Evidence Phospho-specific antibodies, in vitro kinase assay, siRNA/dominant-negative NEK2, centrosome separation assay; Drosophila genetic epistasis for PCP

    PMID:27486244 PMID:28041906

    Open questions at the time
    • Structural basis of phospho-DVL interaction with linker proteins not resolved
    • Whether PCP regulation by NEK2 occurs in mammalian tissues unknown
  16. 2017 High

    Showing that NEK2 interacts with hnRNPA1/2 and directly binds PKM pre-mRNA to promote the PKM2 splice isoform revealed an unexpected role in alternative splicing and metabolic reprogramming.

    Evidence TAP-MS, co-IP, RNA immunoprecipitation, siRNA knockdown in myeloma cells

    PMID:28086949

    Open questions at the time
    • Whether NEK2 kinase activity is required for splicing regulation or only scaffolding function not resolved
    • Breadth of NEK2-regulated splicing events beyond PKM unknown
  17. 2017 High

    Demonstrating that NEK2 phosphorylates EZH2 to block its ubiquitin-proteasome degradation established a paradigm in which NEK2 stabilizes oncoproteins by phosphorylation-dependent protection from ubiquitination.

    Evidence Co-IP, kinase-dead mutant, ubiquitination assay, proteasome inhibitor rescue in glioma stem cells, in vivo tumor model

    PMID:28737508

    Open questions at the time
    • EZH2 phosphorylation site(s) responsible for stabilization not mapped
    • Whether this mechanism is tissue-specific not determined
  18. 2018 High

    Identification of p53 as a NEK2 substrate phosphorylated at Ser315, with functional inhibition of p53 transcriptional activity, connected NEK2 to tumor suppressor inactivation.

    Evidence In vitro kinase assay, in vivo phosphorylation, p53 transcriptional reporter assay

    PMID:30266789

    Open questions at the time
    • Whether Ser315 is the sole regulatory site on p53 targeted by NEK2 unknown
    • In vivo tumorigenic consequences of NEK2-p53 axis not fully characterized
  19. 2020 High

    Cryo-EM and biochemical reconstitution of the APC/C-MCC-Nek2A complex resolved the structural basis of how NEK2A binds APC/C in a conformation-specific manner, explaining checkpoint bypass.

    Evidence Cryo-EM structural analysis, in vitro ubiquitination with E2 variants, cellular validation

    PMID:32307883

    Open questions at the time
    • Resolution of the MR-dipeptide binding interface on APC/C at atomic detail incomplete
    • Whether other substrates exploit this checkpoint bypass mechanism unknown
  20. 2021 High

    Discovery that NEK2 phosphorylates PD-L1 at T194/T210 to prevent its ER-lumenal ubiquitination and degradation linked NEK2 kinase activity to immune evasion.

    Evidence Phospho-site mutagenesis, ubiquitination assay, siRNA knockdown, in vivo tumor immunotherapy model

    PMID:34315872

    Open questions at the time
    • E3 ligase responsible for PD-L1 degradation blocked by NEK2 phosphorylation not identified
    • Breadth of NEK2's role in immune checkpoint regulation beyond PD-L1 unknown
  21. 2022 Medium

    Identification of YAP1 (T143 phosphorylation blocking degradation) and NIK (phosphorylation activating NF-κB) as NEK2 substrates extended the kinase's reach into Hippo and NF-κB signaling, while cooperation with STRIPAK complexes to inactivate MST1/2 provided a second mechanism for Hippo pathway suppression.

    Evidence Co-IP, phospho-site mutagenesis, ubiquitination/CHX chase assays, kinase activity assays, in vivo xenografts

    PMID:35031599 PMID:35705994 PMID:36115593

    Open questions at the time
    • NIK phosphorylation site(s) not mapped
    • Whether STRIPAK cooperation requires NEK2 kinase activity or scaffolding unclear
    • Each finding from a single laboratory

Open questions

Synthesis pass · forward-looking unresolved questions
  • A unified structural and systems-level model explaining how NEK2's mitotic centrosomal/kinetochore functions are coordinated with its non-mitotic roles in cilium disassembly, splicing, immune evasion, and signaling pathway regulation remains to be constructed.
  • No comprehensive phosphoproteomics defining the full NEK2 substrate repertoire in a single system
  • How NEK2 activity is differentially directed toward distinct substrate pools in different cell cycle phases is unknown
  • Structural basis for substrate recognition beyond the Hec1 and APC/C interactions is lacking

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 18 GO:0003723 RNA binding 1
Localization
GO:0005815 microtubule organizing center 10 GO:0005694 chromosome 4 GO:0005634 nucleus 1 GO:0005730 nucleolus 1
Pathway
R-HSA-1640170 Cell Cycle 11 R-HSA-162582 Signal Transduction 5 R-HSA-392499 Metabolism of proteins 4 R-HSA-1852241 Organelle biogenesis and maintenance 3 R-HSA-8953854 Metabolism of RNA 2
Partners
C-NAP1CTNNB1DVLHEC1KIF24MAD1MST2PPP1CA
Complex memberships
Mst2–hSav1–NEK2A complexNEK2 homodimerNEK2A–PP1α complex

Evidence

Reading pass · 46 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1999 Nek2 forms homodimers via an unusual leucine zipper motif in its C-terminal non-catalytic domain; dimerization is required for trans-autophosphorylation and full kinase activity on exogenous substrates. Glycerol gradient centrifugation, yeast two-hybrid, co-precipitation, in vitro kinase assay with leucine zipper deletion mutants The Journal of biological chemistry High 10347187
2001 Nek2A is destroyed at mitotic entry via APC/C-Cdc20-mediated proteasomal degradation, dependent on an extended D-box motif and a KEN-box in its C-terminus; Nek2B is not degraded by this mechanism. Cell synchronization, proteasome inhibitor treatment, APC/C ubiquitination assays, deletion and point mutagenesis of destruction motifs The EMBO journal High 11742988
2002 Nek2 localizes dynamically to centrosomes, chromosomes (prophase to metaphase), and the midbody (telophase), indicating roles in multiple mitotic events beyond centrosome separation. Immunofluorescence microscopy of mitotic cells in culture and in vivo Biochemical and biophysical research communications Medium 11785960
2002 Nek2A and Nek2B are splice variants arising from alternative polyadenylation; both localize to centrosomes and can form homo- and hetero-dimers, but only Nek2A induces centrosome splitting upon overexpression. RT-PCR sequencing, antisense oligonucleotide knockdown, immunoblotting, immunofluorescence, overexpression assays The Biochemical journal High 11742531
2002 The MAPK/Erk1-p90Rsk2 pathway activates Nek2 kinase during meiotic G2/M progression in mouse spermatocytes; p90Rsk2 phosphorylates the Nek2 kinase domain in vitro. MEK inhibitor (U0126) treatment, okadaic acid-induced meiotic progression, in vitro kinase assay with recombinant p90Rsk2 and Nek2 Development (Cambridge, England) High 11923207
2003 Nek2A kinase activity is required for sustained centriole splitting and bipolar spindle formation; kinase-dead Nek2A causes monopolar spindles with unseparated poles, chromosome segregation defects, and cytokinesis failure. Tetracycline-inducible overexpression of wild-type and kinase-dead Nek2A, live cell imaging, immunofluorescence Molecular biology of the cell High 12857871
2003 Nek2 directly interacts with and phosphorylates HMGA2 in mouse pachytene spermatocytes; phosphorylation decreases HMGA2 affinity for DNA and promotes its release from chromatin at the G2/M transition. In vitro binding assay, in vitro kinase assay, co-immunoprecipitation, immunofluorescence, MAPK pathway inhibition Molecular biology of the cell High 14668482
2004 NEK2A interacts with spindle checkpoint protein MAD1 via MAD1's C-terminal leucine-zipper domain; NEK2A localizes to kinetochores and is required for MAD2 (but not MAD1/BUB1/HEC1) recruitment to kinetochores and for proper spindle checkpoint signaling. Co-IP in vitro and in vivo, siRNA knockdown, immunofluorescence, nocodazole challenge assay The Journal of biological chemistry High 14978040
2004 Nek2A regulates centrosomal localization of numatrin (nucleophosmin/B23); Nek2A phosphorylates numatrin in vitro and in vivo, and siRNA depletion of Nek2A delocalizes numatrin from centrosomes. Reciprocal co-immunoprecipitation, GST pulldown, immunofluorescence, siRNA knockdown, in vitro kinase assay FEBS letters High 15388344
2004 Nek2A binds specifically to Erk2 via a conserved docking site in the Nek2A C-terminus; Nek2A is required for centrosomal localization of Erk2, while Erk2 activity is not required for Nek2A localization. Co-IP, immunofluorescence, RNAi depletion, kinase inhibitor treatment Biochemical and biophysical research communications Medium 15358203
2004 Nek2A is activated and interacts with Nek11 at nucleoli in G1/S-arrested cells; Nek2A phosphorylates Nek11 in its C-terminal non-catalytic region, dissociating an autoinhibitory intramolecular interaction and elevating Nek11 kinase activity. Co-immunoprecipitation, in vitro kinase assay, domain mapping, cell synchronization The Journal of biological chemistry High 15161910
2005 Nek2 trafficking to the centrosome requires microtubule-based transport, a C-terminal microtubule-binding/centrosome-localization motif, and the centriolar satellite component PCM-1; centrosomal Nek2 turns over rapidly (t½ ~3 s) and local proteasomal degradation is required for continued recruitment. FRAP, microtubule depolymerization, PCM-1 siRNA depletion, motif mutagenesis, proteasome inhibitor treatment Molecular biology of the cell High 15659651
2005 HEF1 associates with and controls activation of Nek2 at the centrosome; HEF1 depletion causes centrosomal splitting and hyperactivation of Nek2, placing HEF1 as a negative regulator upstream of Nek2. Co-immunoprecipitation, siRNA depletion, kinase activity assays, immunofluorescence Nature cell biology High 16184168
2005 Nlp (ninein-like protein) is phosphorylated by Nek2 kinase at the G2/M transition, causing premature displacement from interphase centrosomes; Nek2 phosphorylation of Nlp primes it for subsequent phosphorylation by Plk1, and both kinases coordinately regulate Nlp displacement. In vitro kinase assay, overexpression of active/kinase-dead Nek2 and Plk1, immunofluorescence, co-immunoprecipitation Molecular and cellular biology High 15684383
2006 Nek2A destruction in prometaphase depends on a C-terminal methionine-arginine (MR) dipeptide that allows Nek2A to bind directly to the APC/C independently of the adaptor Cdc20, rendering its degradation insensitive to the spindle assembly checkpoint. APC/C pull-down assays, C-terminal truncation and MR-mutation, in vitro ubiquitination, Cdc20 immunodepletion Nature cell biology High 16648845
2006 Crystal structure of the Nek2 kinase domain at 2.2 Å reveals an inhibitory helical motif within the activation loop that acts as a steric barrier to active-state formation; autophosphorylation sites identified by mass spectrometry show complex positive and negative regulatory effects on kinase activity correlated with centrosomal splitting in vivo. X-ray crystallography, mass spectrometry identification of autophosphorylation sites, mutational analysis, centrosome splitting assay The Journal of biological chemistry High 17197699
2007 PP1α (but not PP1γ) binds Nek2 via a specific PP1-binding motif outside the kinase domain, dephosphorylates Nek2, and is required for ionizing radiation-induced inhibition of centrosome splitting; ATM activates PP1 to suppress Nek2 activity after DNA damage. Co-immunoprecipitation, siRNA depletion of PP1α, Nek2 PP1-binding motif mutant overexpression, centrosome splitting assay, kinase activity assay Cancer research High 17283141
2007 Beta-catenin is a substrate of Nek2 kinase; beta-catenin is phosphorylated by Nek2, forms a complex with Rootletin at the intercentrosomal linker, and is required for centrosome separation when Nek2 activity increases at mitosis. In vitro kinase assay, co-immunoprecipitation, immunofluorescence, electron microscopy, RNAi knockdown, centrosome splitting assay Genes & development High 18086858
2007 Alternative splicing generates a third Nek2 variant, Nek2C, with a strong bipartite nuclear localization sequence (NLS) spanning the splice site; Nek2C is predominantly nuclear, Nek2B is cytoplasmic, and Nek2A is distributed in both compartments. NLS mutagenesis confirms the differential localization. RT-PCR, mutagenesis of NLS, immunofluorescence, cell fractionation The Journal of biological chemistry High 17626005
2007 NEK2A phosphorylates human Sgo1 at Ser14 and Ser507 in vitro; NEK2A co-localizes with HsSgo1 at kinetochores and phosphorylation of Sgo1 is essential for chromosome congression and proper kinetochore–microtubule attachment. In vitro kinase assay with 32P incorporation, co-IP, immunofluorescence, non-phosphorylatable mutant expression Cell research High 17621308
2007 NIP2/centrobin is a novel substrate of Nek2; Nek2 phosphorylates centrobin and promotes its redistribution from centrosomal aggregates to microtubules, and centrobin knockdown reduces microtubule-organizing activity and causes spindle assembly defects. In vitro kinase assay, immunofluorescence, siRNA knockdown, centrosome/microtubule co-localization Journal of cell science High 17535851
2008 NEK2A phosphorylates Hec1 at Ser165 during mitosis; non-phosphorylatable Hec1(S165A) causes increased microtubule attachment errors (syntelic/monotelic); in vitro, Nek2A phosphorylation of Hec1 increases the affinity of the Ndc80 complex for microtubules. In vitro kinase assay, phospho-specific antibody, expression of non-phosphorylatable mutant, in vitro microtubule-binding reconstitution, immunofluorescence Oncogene High 18297113
2008 Crystal structures of wild-type Nek2 kinase domain (1.55 Å, ADP-bound), T175A apo form, and ATP-analogue-bound form reveal multiple conformations around the nucleotide-binding site and suggest mechanisms for auto-inhibition involving activation loop dynamics. X-ray crystallography, site-directed mutagenesis of identified auto-inhibitory residues Journal of molecular biology High 19124027
2009 Both Mad2 and Cdc20 physically associate with Nek2 and can be phosphorylated by Nek2 in vitro; Nek2 overexpression enhances Mad2-induced mitotic delay, suggesting Nek2 acts on the Mad2-Cdc20 complex to regulate the mitotic checkpoint. Co-immunoprecipitation, in vitro kinase assay, overexpression phenotype assay Experimental and molecular pathology Medium 20034488
2010 Hippo pathway components Mst2 and hSav1 directly interact with Nek2A and regulate its centrosomal localization and ability to phosphorylate C-Nap1 and rootletin; this hSav1-Mst2-Nek2A pathway acts complementarily to Eg5 in centrosome disjunction and bipolar spindle formation. Co-immunoprecipitation, siRNA knockdown, immunofluorescence, kinase activity assay, Eg5 inhibitor epistasis Nature cell biology High 21076410
2011 Plk1 functions upstream of the Mst2-Nek2A module by phosphorylating Mst2; unphosphorylated Mst2 promotes assembly of Nek2A-PP1γ-Mst2 complexes that suppress Nek2A activity, while Plk1-phosphorylated Mst2 prevents PP1γ from binding and allows Nek2A to drive centrosome disjunction. Co-immunoprecipitation, kinase activity assays, Plk1 inhibition/depletion, centrosome disjunction assay Current biology : CB High 21723128
2011 The Nek2 leucine zipper exists in two conformations of nearly equal population that interconvert at ~17 s⁻¹, corresponding to two heptad register frames; a cysteine mutant locks one conformation and validates the model. NMR spectroscopy (15N HSQC, exchange spectra), site-directed mutagenesis The Journal of biological chemistry High 21669869
2011 Irreversible inhibition of Nek2 kinase can be achieved by cysteine-targeted compounds acting on Cys22 near the glycine-rich loop; compound JH295 (propynamide 16) irreversibly inactivates Nek2 in cells without affecting Cdk1, Aurora B, or Plk1. Biochemical kinase assays, cell-based kinase activity assay, selectivity profiling against mitotic kinases Journal of medicinal chemistry High 21627121
2015 Nek2 phosphorylates Kif24, a microtubule-depolymerizing kinesin, stimulating its depolymerizing activity and suppressing cilia outgrowth in proliferating cells; co-localization of Nek2 and Kif24 at the S/G2 phase drives cilium disassembly independently of Aurora A and HDAC6. Co-immunoprecipitation, in vitro kinase assay, siRNA knockdown, immunofluorescence, ciliation assay, rescue experiments Nature communications High 26290419
2015 Kendrin (pericentrin) associates with Nek2A, inhibits its kinase activity in vitro in a dose-dependent manner, and ectopic expression of the Nek2A-binding region of kendrin reduces premature centrosome splitting. Co-immunoprecipitation, in vitro kinase inhibition assay, overexpression, centrosome splitting assay Biochemical and biophysical research communications Medium 20599736
2015 Nek2 overexpression promotes cilia resorption in cultured cells via the tubulin deacetylase Hdac6; Nek2 knockdown causes centriole defects at the left-right organizer; Nek2 interaction with the nucleoporin Nup98 regulates cilium resorption. Xenopus nek2 morpholino knockdown and overexpression, immunofluorescence, co-IP, HDAC6 inhibitor rescue, ciliation assay Development (Cambridge, England) Medium 26493400
2015 Cep68 is phosphorylated by Nek2 in vivo, and this phosphorylation promotes Cep68 degradation at mitosis; the SCF-βTrCP complex is responsible for mitotic Cep68 destruction. In vivo phosphorylation assay, immunoprecipitation, SCF complex inhibition, cell cycle fractionation European journal of cell biology Medium 25704143
2016 Dishevelled (DVL) is a Nek2 substrate; Nek2 phosphorylates DVL at multiple residues creating binding interfaces for centrosomal linker proteins CDK5RAP2 and C-NAP1; DVL displacement from the centrosome upon NEK2 phosphorylation is coupled to linker dissolution and centrosome separation. Co-IP, phospho-specific antibodies, in vitro kinase assay, siRNA knockdown, centrosome separation assay, dominant-negative Nek2 Proceedings of the National Academy of Sciences of the United States of America High 27486244
2016 In Drosophila, APC/CFzr/Cdh1 negatively regulates Nek2 kinase levels, and Nek2 phosphorylates Dishevelled (Dsh) to promote its degradation, thereby controlling Dsh levels required for planar cell polarity establishment. Genetic epistasis in Drosophila PCP assay, APC/C loss-of-function, Nek2 overexpression, Dsh protein level measurement Developmental cell Medium 28041906
2017 NEK2 interacts with hnRNPA1/2 and binds to PKM pre-mRNA intronic sequence flanking exon 9, promoting alternative splicing of PKM to increase the PKM2/PKM1 ratio and aerobic glycolysis in multiple myeloma cells. TAP-MS, co-immunoprecipitation, RNA immunoprecipitation (RIP), ChIP-PCR, Western blot, siRNA knockdown Journal of hematology & oncology High 28086949
2017 NEK2 forms a complex with EZH2 in glioma stem cells; NEK2-mediated phosphorylation of EZH2 protects it from ubiquitin-proteasome degradation, thereby stabilizing EZH2 protein in a NEK2 kinase activity-dependent manner. Co-immunoprecipitation, proteasome inhibitor rescue, kinase-dead NEK2 mutant, EZH2 ubiquitination assay, in vivo tumor model The Journal of clinical investigation High 28737508
2018 NEK2 phosphorylates p53 at Ser315 in vitro and in vivo and functionally inhibits p53 transcriptional activity. In vitro kinase assay, in vivo phosphorylation, p53 transcriptional reporter assay Proceedings of the National Academy of Sciences of the United States of America High 30266789
2020 Nek2A binds the APC/C-MCC complex in a conformationally specific manner and can be ubiquitinated efficiently by APC/C together with either UbcH10 or UbcH5 E2 enzymes; this unique binding mode allows prometaphase-specific ubiquitination even when the SAC is active. Biochemical reconstitution of APC/C-MCC-Nek2A complex, cryo-EM structural analysis, in vitro ubiquitination assays with E2 variants, cellular functional validation EMBO reports High 32307883
2021 NEK2 phosphorylates PD-L1 at T194 and T210 within a NEK binding motif (F/LXXS/T) in the glycosylation-rich region, preventing ubiquitin-proteasome-mediated degradation of PD-L1 in the ER lumen and thereby stabilizing surface PD-L1 expression. Co-immunoprecipitation, phosphorylation site mutagenesis, ubiquitination assay, immunofluorescence, siRNA knockdown, in vivo tumor immunotherapy model Nature communications High 34315872
2021 NEK2 modulates an RBFOX2-dependent pro-mesenchymal alternative splicing program in triple-negative breast cancer cells; NEK2 depletion reduces RBFOX2 expression and induces epithelial phenotypic reversion. RNA-seq, siRNA knockdown, bioinformatics, RBFOX2 rescue experiments, cell morphology and motility assays Journal of experimental & clinical cancer research : CR Medium 34930366
2022 NEK2 phosphorylates YAP1 at Thr-143, protecting YAP1 from ubiquitination-dependent proteasomal degradation; NEK2-mediated YAP1 stabilization promotes ESCC migration and invasion. Co-immunoprecipitation, cycloheximide chase, ubiquitination assay, phospho-site mutagenesis (T143A), overexpression/knockdown rescue, in vivo xenograft Cell communication and signaling : CCS High 35705994
2022 NEK2 activates the non-canonical NF-κB pathway by phosphorylating NIK, increasing NIK activity and stability; NEK2 knockdown suppresses GBM proliferation and invasion by reducing NIK/NF-κB signaling. RNA-seq, co-immunoprecipitation, kinase activity assay, NIK phosphorylation assay, siRNA knockdown, in vivo xenograft Cell death & disease Medium 35031599
2022 NEK2 cooperates with STRIPAK complexes to dephosphorylate MST1/2, thereby inactivating the Hippo signaling cascade and increasing nuclear YAP activity in cervical cancer cells. Mass spectrometry, bilateral co-immunoprecipitation, YAP phosphorylation assay, siRNA/overexpression, in vivo xenograft Cancer letters Medium 36115593
2013 Nek2 directly binds and phosphorylates TRF1 at multiple sites; TRF1 is required for Nek2-overexpression-induced centrosome amplification, multinucleation, and chromosomal instability in breast cancer cells. Co-immunoprecipitation, in vitro kinase assay, siRNA depletion of TRF1, overexpression epistasis, immunofluorescence centrosome counting Cell cycle (Georgetown, Tex.) Medium 24091727
2014 Small-molecule INH compounds bind Hec1 at residues W395/L399/K400 (aa 394-408), blocking NEK2-mediated Hec1 S165 phosphorylation; INH binding to Hec1 simultaneously presents the NEK2 D-box region for APC/C-mediated proteasomal degradation of NEK2 via a 'death-trap' mechanism. In vitro kinase assay, co-immunoprecipitation domain mapping, Nek2 R361L D-box mutant, ubiquitination/degradation assay, in vivo tumor model Oncogene High 24662830
2019 Nek2 binds beta-catenin in sorafenib-treated HCC cells, stabilizes beta-catenin protein, and promotes its nuclear translocation, thereby activating Wnt target gene transcription and conferring sorafenib resistance. Co-immunoprecipitation, Western blot (protein stability), nuclear fractionation, siRNA knockdown, in vivo xenograft Journal of experimental & clinical cancer research : CR Medium 31319849

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2003 Nek2A kinase stimulates centrosome disjunction and is required for formation of bipolar mitotic spindles. Molecular biology of the cell 212 12857871
2005 The focal adhesion scaffolding protein HEF1 regulates activation of the Aurora-A and Nek2 kinases at the centrosome. Nature cell biology 202 16184168
2007 beta-Catenin is a Nek2 substrate involved in centrosome separation. Genes & development 197 18086858
2001 APC/C-mediated destruction of the centrosomal kinase Nek2A occurs in early mitosis and depends upon a cyclin A-type D-box. The EMBO journal 183 11742988
2002 The Nek2 protein kinase: a novel regulator of centrosome structure. Oncogene 181 12214248
2010 Components of the Hippo pathway cooperate with Nek2 kinase to regulate centrosome disjunction. Nature cell biology 153 21076410
2005 Nek2 kinase in chromosome instability and cancer. Cancer letters 145 16084011
2015 Nek2 activation of Kif24 ensures cilium disassembly during the cell cycle. Nature communications 136 26290419
2006 Early mitotic degradation of Nek2A depends on Cdc20-independent interaction with the APC/C. Nature cell biology 134 16648845
2017 NEK2 Promotes Aerobic Glycolysis in Multiple Myeloma Through Regulating Splicing of Pyruvate Kinase. Journal of hematology & oncology 126 28086949
2011 Plk1 controls the Nek2A-PP1γ antagonism in centrosome disjunction. Current biology : CB 112 21723128
2021 NEK2 inhibition triggers anti-pancreatic cancer immunity by targeting PD-L1. Nature communications 110 34315872
2004 NEK2A interacts with MAD1 and possibly functions as a novel integrator of the spindle checkpoint signaling. The Journal of biological chemistry 106 14978040
2005 Dynamic recruitment of Nek2 kinase to the centrosome involves microtubules, PCM-1, and localized proteasomal degradation. Molecular biology of the cell 98 15659651
2013 Role of Nek2 on centrosome duplication and aneuploidy in breast cancer cells. Oncogene 93 23708664
2006 Structure and regulation of the human Nek2 centrosomal kinase. The Journal of biological chemistry 90 17197699
2018 CircRNA circPDSS1 promotes the gastric cancer progression by sponging miR-186-5p and modulating NEK2. Journal of cellular physiology 87 30417526
2007 Nek2 as an effective target for inhibition of tumorigenic growth and peritoneal dissemination of cholangiocarcinoma. Cancer research 82 17942892
1999 Activity of the human centrosomal kinase, Nek2, depends on an unusual leucine zipper dimerization motif. The Journal of biological chemistry 82 10347187
2005 Coordinate regulation of the mother centriole component nlp by nek2 and plk1 protein kinases. Molecular and cellular biology 80 15684383
2009 An essential role for the Plasmodium Nek-2 Nima-related protein kinase in the sexual development of malaria parasites. The Journal of biological chemistry 79 19491095
2007 Protein phosphatase-1alpha regulates centrosome splitting through Nek2. Cancer research 76 17283141
2003 Phosphorylation of high-mobility group protein A2 by Nek2 kinase during the first meiotic division in mouse spermatocytes. Molecular biology of the cell 76 14668482
2007 Characterization of NIP2/centrobin, a novel substrate of Nek2, and its potential role in microtubule stabilization. Journal of cell science 74 17535851
2004 Inhibition of centrosome separation after DNA damage: a role for Nek2. Radiation research 74 15387139
2013 Upregulation of NEK2 is associated with drug resistance in ovarian cancer. Oncology reports 73 24337664
2002 The MAPK pathway triggers activation of Nek2 during chromosome condensation in mouse spermatocytes. Development (Cambridge, England) 73 11923207
2000 Nek2B, a novel maternal form of Nek2 kinase, is essential for the assembly or maintenance of centrosomes in early Xenopus embryos. The EMBO journal 73 10775266
2011 Irreversible Nek2 kinase inhibitors with cellular activity. Journal of medicinal chemistry 72 21627121
2017 Targeting NEK2 attenuates glioblastoma growth and radioresistance by destabilizing histone methyltransferase EZH2. The Journal of clinical investigation 71 28737508
2010 The Ras oncogene signals centrosome amplification in mammary epithelial cells through cyclin D1/Cdk4 and Nek2. Oncogene 70 20581865
2020 Targeting NEK2 impairs oncogenesis and radioresistance via inhibiting the Wnt1/β-catenin signaling pathway in cervical cancer. Journal of experimental & clinical cancer research : CR 69 32907622
2014 Overexpression of the Nek2 kinase in colorectal cancer correlates with beta-catenin relocalization and shortened cancer-specific survival. Journal of surgical oncology 69 25043295
2008 The mitotic checkpoint kinase NEK2A regulates kinetochore microtubule attachment stability. Oncogene 69 18297113
2002 Alternative splice variants of the human centrosome kinase Nek2 exhibit distinct patterns of expression in mitosis. The Biochemical journal 67 11742531
2010 Novel combination treatment for colorectal cancer using Nek2 siRNA and cisplatin. Cancer science 66 20345485
2020 Circular RNA circTNPO3 Regulates Paclitaxel Resistance of Ovarian Cancer Cells by miR-1299/NEK2 Signaling Pathway. Molecular therapy. Nucleic acids 64 32791450
2007 Alternative splicing controls nuclear translocation of the cell cycle-regulated Nek2 kinase. The Journal of biological chemistry 57 17626005
2018 NEK2 promotes hepatocellular carcinoma migration and invasion through modulation of the epithelial-mesenchymal transition. Oncology reports 55 29399700
2016 Dishevelled is a NEK2 kinase substrate controlling dynamics of centrosomal linker proteins. Proceedings of the National Academy of Sciences of the United States of America 54 27486244
2014 Novel small molecules disrupting Hec1/Nek2 interaction ablate tumor progression by triggering Nek2 degradation through a death-trap mechanism. Oncogene 52 24662830
2020 Circular RNA circPITX1 knockdown inhibits glycolysis to enhance radiosensitivity of glioma cells by miR-329-3p/NEK2 axis. Cancer cell international 49 32190004
2013 Nek2-targeted ASO or siRNA pretreatment enhances anticancer drug sensitivity in triple‑negative breast cancer cells. International journal of oncology 47 23340795
2010 Aminopyrazine inhibitors binding to an unusual inactive conformation of the mitotic kinase Nek2: SAR and structural characterization. Journal of medicinal chemistry 47 20936789
2019 Nek2 augments sorafenib resistance by regulating the ubiquitination and localization of β-catenin in hepatocellular carcinoma. Journal of experimental & clinical cancer research : CR 46 31319849
2009 Synthesis and biological evaluation of a series of novel inhibitor of Nek2/Hec1 analogues. Journal of medicinal chemistry 46 19243176
2008 Insights into the conformational variability and regulation of human Nek2 kinase. Journal of molecular biology 46 19124027
2007 Phosphorylation of human Sgo1 by NEK2A is essential for chromosome congression in mitosis. Cell research 46 17621308
2019 NEK2 promotes proliferation, migration and tumor growth of gastric cancer cells via regulating KDM5B/H3K4me3. American journal of cancer research 45 31815040
2002 Nek2 localizes to multiple sites in mitotic cells, suggesting its involvement in multiple cellular functions during the cell cycle. Biochemical and biophysical research communications 45 11785960
2013 Centrosomal kinase Nek2 cooperates with oncogenic pathways to promote metastasis. Oncogenesis 43 24018644
2017 Hepatoma cell functions modulated by NEK2 are associated with liver cancer progression. International journal of cancer 41 27925179
2015 Role of NEK2A in human cancer and its therapeutic potentials. BioMed research international 41 25705694
2020 microRNA-128-3p overexpression inhibits breast cancer stem cell characteristics through suppression of Wnt signalling pathway by down-regulating NEK2. Journal of cellular and molecular medicine 40 32558224
2013 Cdk4 and nek2 signal binucleation and centrosome amplification in a her2+ breast cancer model. PloS one 38 23776583
2009 Nek2 targets the mitotic checkpoint proteins Mad2 and Cdc20: a mechanism for aneuploidy in cancer. Experimental and molecular pathology 38 20034488
1999 Two structural variants of Nek2 kinase, termed Nek2A and Nek2B, are differentially expressed in Xenopus tissues and development. Developmental biology 38 10191058
2018 Literature-based automated discovery of tumor suppressor p53 phosphorylation and inhibition by NEK2. Proceedings of the National Academy of Sciences of the United States of America 36 30266789
2015 The NIMA-like kinase Nek2 is a key switch balancing cilia biogenesis and resorption in the development of left-right asymmetry. Development (Cambridge, England) 36 26493400
2022 Genetic Aberrations and Interaction of NEK2 and TP53 Accelerate Aggressiveness of Multiple Myeloma. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 35 35088582
2013 Mitotic perturbations induced by Nek2 overexpression require interaction with TRF1 in breast cancer cells. Cell cycle (Georgetown, Tex.) 34 24091727
2015 Effect of silencing NEK2 on biological behaviors of HepG2 in human hepatoma cells and MAPK signal pathway. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine 33 26337275
2020 LIN9 and NEK2 Are Core Regulators of Mitotic Fidelity That Can Be Therapeutically Targeted to Overcome Taxane Resistance. Cancer research 32 32054769
2011 An undecided coiled coil: the leucine zipper of Nek2 kinase exhibits atypical conformational exchange dynamics. The Journal of biological chemistry 32 21669869
2011 Abnormal expression of Nek2 and β-catenin in breast carcinoma: clinicopathological correlations. Histopathology 32 22014044
2004 Nucleolar Nek11 is a novel target of Nek2A in G1/S-arrested cells. The Journal of biological chemistry 32 15161910
2016 Molecular modeling and docking of small molecule inhibitors against NEK2. Bioinformation 31 28104962
2016 Increased NEK2 in hepatocellular carcinoma promotes cancer progression and drug resistance by promoting PP1/Akt and Wnt activation. Oncology reports 30 27509921
2010 Cyclin A and Nek2A: APC/C-Cdc20 substrates invisible to the mitotic spindle checkpoint. Biochemical Society transactions 30 20074038
2005 Live cell imaging reveals distinct roles in cell cycle regulation for Nek2A and Nek2B. Biochimica et biophysica acta 30 15950749
2021 The oncogenic kinase NEK2 regulates an RBFOX2-dependent pro-mesenchymal splicing program in triple-negative breast cancer cells. Journal of experimental & clinical cancer research : CR 29 34930366
2020 High expression of NEK2 promotes gastric cancer progression via activating AKT signaling. Journal of physiology and biochemistry 29 33201407
2022 NEK2 enhances malignancies of glioblastoma via NIK/NF-κB pathway. Cell death & disease 28 35031599
2020 Exosome-Derived miR-486-5p Regulates Cell Cycle, Proliferation and Metastasis in Lung Adenocarcinoma via Targeting NEK2. Frontiers in bioengineering and biotechnology 28 32322578
2019 Simulated microgravity inhibits the viability and migration of glioma via FAK/RhoA/Rock and FAK/Nek2 signaling. In vitro cellular & developmental biology. Animal 28 30820814
2024 Role of NEK2 in tumorigenesis and tumor progression. Trends in molecular medicine 25 39181803
2022 RPL17 Promotes Colorectal Cancer Proliferation and Stemness through ERK and NEK2/β-catenin Signaling Pathways. Journal of Cancer 25 35711835
2019 microRNA-138 induces cell survival and reduces WNT/β-catenin signaling of osteoarthritis chondrocytes through NEK2. IUBMB life 25 31034758
2012 Nek2A contributes to tumorigenic growth and possibly functions as potential therapeutic target for human breast cancer. Journal of cellular biochemistry 25 22234886
2004 Suppression of Nek2A in mouse early embryos confirms its requirement for chromosome segregation. Journal of cell science 25 15479717
2004 Nek2A kinase regulates the localization of numatrin to centrosome in mitosis. FEBS letters 24 15388344
2022 One shoot, three birds: Targeting NEK2 orchestrates chemoradiotherapy, targeted therapy, and immunotherapy in cancer treatment. Biochimica et biophysica acta. Reviews on cancer 23 35157980
2022 NEK2 promotes the migration and proliferation of ESCC via stabilization of YAP1 by phosphorylation at Thr-143. Cell communication and signaling : CCS 23 35705994
2022 NEK2 inactivates the Hippo pathway to advance the proliferation of cervical cancer cells by cooperating with STRIPAK complexes. Cancer letters 23 36115593
2017 MiR-486-5p negatively regulates oncogenic NEK2 in hepatocellular carcinoma. Oncotarget 22 28881785
2014 Nek2 is a novel regulator of B cell development and immunological response. BioMed research international 22 25485281
2010 Involvement of a centrosomal protein kendrin in the maintenance of centrosome cohesion by modulating Nek2A kinase activity. Biochemical and biophysical research communications 22 20599736
2021 Silencing of Nek2 suppresses the proliferation, migration and invasion and induces apoptosis of breast cancer cells by regulating ERK/MAPK signaling. Journal of molecular histology 21 34009515
2015 Cep68 can be regulated by Nek2 and SCF complex. European journal of cell biology 21 25704143
2014 Therapeutic melting pot of never in mitosis gene a related kinase 2 (Nek2): a perspective on Nek2 as an oncology target and recent advancements in Nek2 small molecule inhibition. Journal of medicinal chemistry 21 24517277
2020 A unique binding mode of Nek2A to the APC/C allows its ubiquitination during prometaphase. EMBO reports 20 32307883
2020 lncRNA SNHG1 Promotes Progression of Cervical Cancer Through miR-195/NEK2 Axis. Cancer management and research 20 33204155
2016 High NEK2 Expression Is a Predictor of Tumor Recurrence in Hepatocellular Carcinoma Patients After Hepatectomy. Anticancer research 20 26851035
2016 APC/CFzr/Cdh1-Dependent Regulation of Planar Cell Polarity Establishment via Nek2 Kinase Acting on Dishevelled. Developmental cell 20 28041906
2012 Detection of IGF2BP3, HOXB7, and NEK2 mRNA expression in brush cytology specimens as a new diagnostic tool in patients with biliary strictures. PloS one 20 22879911
2014 Inhibition of Nek2 by small molecules affects proteasome activity. BioMed research international 19 25313354
2013 Model system for irreversible inhibition of Nek2: thiol addition to ethynylpurines and related substituted heterocycles. Organic & biomolecular chemistry 19 24213855
2004 Nek2A specifies the centrosomal localization of Erk2. Biochemical and biophysical research communications 19 15358203
2021 NEK2 plays an active role in Tumorigenesis and Tumor Microenvironment in Non-Small Cell Lung Cancer. International journal of biological sciences 18 34131401
2018 Importance of protein flexibility on molecular recognition: modeling binding mechanisms of aminopyrazine inhibitors to Nek2. Physical chemistry chemical physics : PCCP 18 29270587