| 1992 |
NEK1 (Nek1) is a dual-specificity kinase that phosphorylates substrates on serine, threonine, AND tyrosine residues in vitro; its kinase domain expressed in bacteria phosphorylated exogenous substrates (preferentially beta-casein) on all three hydroxyamino acids. The kinase domain is most similar (42% identity) to the catalytic domain of NIMA, and like NIMA, Nek1 has a long basic C-terminal extension. |
In vitro kinase assay with bacterial expression of kinase domain; substrate phosphorylation on Ser/Thr/Tyr characterized |
The EMBO journal |
High |
1382974
|
| 2000 |
Loss-of-function mutations in Nek1 (partial internal deletion in kat; single-base insertion in kat2J) cause polycystic kidney disease and pleiotropic phenotypes (facial dysmorphism, dwarfing, male sterility, anemia, cystic choroid plexus) in mice, establishing that Nek1 participates in multiple signaling pathways regulating diverse cellular processes including kidney tubular homeostasis. |
Positional cloning, mutant mouse genetic analysis, molecular characterization of mutations |
Proceedings of the National Academy of Sciences of the United States of America |
High |
10618398
|
| 2003 |
The central coiled-coil region of the human NEK1 regulatory domain interacts with KIF3A (a motor protein involved in PKD), tuberin, alpha-catulin, and proteins involved in DNA double-strand break repair and neural development, as identified by yeast two-hybrid screening of a fetal brain cDNA library. |
Yeast two-hybrid screen using NEK1 kinase and regulatory domains as baits; domain mapping of interactions to central coiled-coil region |
Biochemistry |
Medium |
14690447
|
| 2006 |
Full-length murine Nek1 localizes to the cytoplasm despite containing two classical nuclear localization signals, with the C-terminus bearing cytoplasmic localization signals. Ectopic overexpression of nuclear-targeted Nek1 constructs causes abnormal chromatin condensation (DNA confined to nuclear periphery) and nuclear pore complex dispersal, without mitotic or apoptotic markers—phenotypically similar to NIMA overexpression. |
Subcellular fractionation, immunofluorescence, ectopic overexpression of nuclear-targeted constructs in mammalian cells |
Biochimica et biophysica acta |
Medium |
16603261
|
| 2008 |
Nek1 localizes to the basal body region of primary cilia. Overexpression of full-length Nek1 inhibits ciliogenesis in IMCD3 cells without disrupting centrosomes. Overexpression of the kinase domain alone (lacking the acidic regulatory domain) causes centrosome loss and secondary ciliogenesis failure. Both effects require kinase activity and the coiled-coil domain. A ciliary targeting region was defined within the coiled-coil domain. |
Immunofluorescence localization, overexpression/kinase-dead mutant analysis in MDCK and IMCD3 cells, kinase-inactivating mutations |
BMC cell biology |
High |
18387364 18533026
|
| 2009 |
Nek1 associates with VDAC1 (voltage-dependent anion channel 1) at mitochondria and phosphorylates VDAC1 at Ser193. Loss of Nek1 or expression of kinase-dead Nek1 results in loss of VDAC1-S193 phosphorylation, exaggerated mitochondrial membrane permeability, and accelerated cell death. A phospho-mimic VDAC1-S193E rescues excessive MMP and transiently keeps cells alive after DNA damage; phospho-deficient VDAC1-S193A causes constitutive cell death. |
Yeast two-hybrid, GST pull-down, reciprocal co-immunoprecipitation, in vitro kinase assay, ectopic expression of phospho-mimic/phospho-deficient VDAC1 mutants, mitochondrial permeability assays |
Cell cycle (Georgetown, Tex.) |
High |
19158487
|
| 2009 |
NEK1 contains functional nuclear localization signals and a nuclear export signal-dependent export pathway, cycling through the nucleus. This nucleocytoplasmic shuttling suggests NEK1 can transduce signals from the ciliary-basal body region to nuclear gene expression. |
Nuclear export signal inhibitor treatment, fluorescence localization, functional NLS/NES identification |
Biochemical and biophysical research communications |
Medium |
19699716
|
| 2010 |
Phosphorylation of VDAC1 at Ser193 by Nek1 closes the VDAC1 channel and prevents cytochrome c efflux. Unphosphorylated (wild-type or S193A) VDAC1 adopts an open configuration and allows cytochrome c release; phospho-mimic S193E VDAC1 remains closed and prevents cytochrome c leakage in liposome reconstitution assays. |
Atomic force microscopy (AFM) of VDAC1 channel conformation, liposome-based cytochrome c conductance assay, in vitro kinase phosphorylation, phospho-mutant (S193A, S193E) constructs |
Biochemical and biophysical research communications |
High |
20230784
|
| 2010 |
Nek1 knockdown causes a delay in DNA repair after treatment with MMS, H2O2, and cisplatin; reduces Chk1 phosphorylation in response to cisplatin; and abrogates the G2/M arrest normally induced by MMS and H2O2 damage, establishing Nek1 as an early actor in the DNA damage response and G2/M checkpoint. |
Stable siRNA knockdown, comet assay, phospho-Chk1 immunoblot, flow cytometry cell cycle analysis |
Mutagenesis |
Medium |
20501547
|
| 2011 |
NEK1-deficient cells undergo major errors in mitotic chromosome segregation and cytokinesis, becoming aneuploid. These cells transform and form tumors in syngeneic mice. NEK1+/− mice develop lymphomas with higher incidence than wild-type, establishing NEK1 as required for genome stability at multiple junctures. |
Loss-of-function mouse genetics, colony transformation assay, tumor injection in syngeneic mice, karyotype analysis |
Molecular cancer |
Medium |
21214959
|
| 2011 |
Nek1 functions independently of ATM and ATR kinases in the DNA damage response: chemical inhibitors or genetic loss of ATM/ATR do not alter Nek1 kinase activity or its localization to nuclear DNA damage foci, and ATM/ATR activities are intact in Nek1-null cells, placing Nek1 in a parallel or upstream DNA damage sensing pathway. |
Chemical inhibitors of ATM/ATR, ATM/ATR genetic knockouts, kinase activity assays, immunofluorescence of damage foci |
Cell cycle (Georgetown, Tex.) |
Medium |
21301226
|
| 2011 |
Loss of NEK1 in murine spermatocytes results in failure to remove cohesin (SMC3) from meiotic chromosome cores at the end of prophase I, causing complete male infertility, demonstrating a role for NEK1 in cohesin redistribution during meiotic prophase I. |
Nek1kat2J mutant mouse analysis, immunostaining of meiotic chromosome spreads for SMC3 cohesin |
Genes |
Medium |
21931878
|
| 2011 |
Nek1 phosphorylates TAZ at a site essential for TAZ's ubiquitination and proteasomal targeting of polycystin 2 (PC2). Loss of Nek1 leads to underphosphorylation of TAZ and abnormal accumulation of PC2. TAZ in turn targets Nek1 for degradation, forming a negative feedback loop linking Nek1 and TAZ to maintain PC2 levels and proper ciliogenesis. |
Co-immunoprecipitation, in vitro kinase assay, mutant expression, western blotting for PC2 levels |
Journal of the American Society of Nephrology : JASN |
Medium |
21474562
|
| 2012 |
Nek1 phosphorylates the VHL tumor suppressor protein (pVHL) at multiple sites including Ser168, leading to pVHL proteasomal degradation. Phosphorylation at Ser168 regulates cilia stability: non-phosphorylatable pVHL (S168A) reconstituted in VHL-deficient cells induces more stable cilia than wild-type VHL, linking Nek1-mediated pVHL phosphorylation to ciliary homeostasis. |
In vitro kinase assay, site-directed mutagenesis (S168A), reconstitution in VHL-deficient cells, ciliation assay |
Cell cycle (Georgetown, Tex.) |
Medium |
23255108
|
| 2013 |
Nek1 associates with ATR-ATRIP complex, is required for maintaining ATRIP protein levels, the ATR-ATRIP association, and basal ATR kinase activity even in undamaged cells. Upon DNA damage, Nek1 is required for efficient phosphorylation of multiple ATR substrates and ATR autophosphorylation at T1989. The pro-ATR activity of Nek1 requires its own kinase activity and its physical interaction with ATR-ATRIP. |
Co-immunoprecipitation, kinase activity assays, siRNA knockdown, epistasis analysis, ATR substrate phosphorylation assays |
Proceedings of the National Academy of Sciences of the United States of America |
High |
23345434
|
| 2013 |
Nek1 interacts with Ku80 and is required for chromatin loading of Ku80 and multiple DNA replication factors during S phase. Nek1 deficiency causes severe S-phase delay, replication stress, and checkpoint activation under normal growth conditions, identifying a role for Nek1 in S-phase progression via Ku80 interaction. |
Co-immunoprecipitation, chromatin fractionation, BrdU incorporation assay, flow cytometry |
Cell cycle (Georgetown, Tex.) |
Medium |
23851348
|
| 2015 |
C21ORF2 physically interacts with NEK1. C21ORF2 depletion impairs homologous recombination (but not NHEJ) after ionizing radiation damage, and this defect is rescued by NEK1 overexpression, placing C21ORF2 in the same DNA damage repair pathway as NEK1. |
Co-immunoprecipitation, siRNA knockdown, HR/NHEJ reporter assays, rescue by NEK1 overexpression |
Acta biochimica et biophysica Sinica |
Medium |
26290490
|
| 2016 |
NEK1 phosphorylates Rad54 at Ser572 specifically in late G2 phase to promote homologous recombination. Nek1 deficiency or unphosphorylatable Rad54-S572A causes unresolved Rad51 foci and HR defects; phospho-mimic Rad54-S572E promotes HR and rescues Nek1-loss HR defects. G2-specific phosphorylation promotes Rad51 removal from chromatin. In S phase, absence of this phosphorylation is required for replication fork stability, as phospho-mimic Rad54 in S phase causes Rad51 removal and fork degradation. |
In vitro kinase assay, phospho-specific antibody, site-directed mutagenesis (S572A, S572E), HR assay, Rad51 foci analysis, replication fork stability assay (DNA fiber assay) |
Molecular cell |
High |
27264870
|
| 2016 |
NEK1 regulates cohesin removal during meiotic prophase I via an NEK1–PDS5B–PP1γ–WAPL axis. NEK1 interacts with PDS5B and phosphorylates PP1γ; PP1γ dephosphorylates WAPL, enabling its retention on chromosome cores and promoting loss of cohesion at the end of prophase I in mammals. |
Co-immunoprecipitation, phospho-substrate analysis, Nek1 knockout mouse, immunostaining of meiotic chromosome spreads |
Cell reports |
High |
27760328
|
| 2016 |
CEP104, a ciliopathy protein involved in cilia formation and length regulation, interacts with NEK1 through NEK1's C2HC zinc finger array. NEK1 competes with the centriole-capping protein CP110 for binding to Cep104, providing a structural link between NEK1 and cilia assembly regulation. |
Structural studies (biochemical), co-immunoprecipitation, competition binding assay between NEK1 and CP110 for Cep104 |
Structure (London, England : 1993) |
Medium |
28017521
|
| 2017 |
TLK1 (Tousled-Like Kinase 1) directly interacts with and phosphorylates NEK1 at Thr141 within the kinase domain, activating NEK1. This TLK1→NEK1→ATR→Chk1 pathway mediates the DNA damage response. The phospho-deficient NEK1-T141A mutant impairs ATR and Chk1 activation after DNA damage and alters cell cycle response to oxidative stress. TLK1-NEK1 co-relocalize to nuclear repair foci with γH2AX after doxorubicin. |
Co-immunoprecipitation, proteomic screen, in vitro kinase assay (TLK1 phosphorylating NEK1-T141), NEK1-T141A mutant overexpression, ATR/Chk1 activation assays, immunofluorescence |
Cell cycle (Georgetown, Tex.) |
High |
28426283
|
| 2017 |
The crystal structure of the NEK1 kinase domain was solved in apo and ATP-mimetic inhibitor-bound forms, revealing structural basis for inhibitor specificity and providing a template for rational drug design. |
X-ray crystallography of NEK1 kinase domain (apo and inhibitor-bound); dynamic protein interactome characterized by proteomics after cisplatin treatment |
Scientific reports |
High |
28710492
|
| 2017 |
NEK1 regulates meiosis I spindle assembly: loss of NEK1 causes elongated and/or multipolar meiotic spindles and abnormal chromosome congression. NEK1 regulates protein levels and localization of α-adducin (ADD1) and myosin X (MYO10) on meiotic spindles. Hyperphosphorylation of ADD1 in NEK1 mutants prevents its interaction with MYO10, disrupting spindle architecture. |
Nek1 knockout mouse, immunostaining of meiotic spindles, co-immunoprecipitation of ADD1-MYO10, western blot of phospho-ADD1 |
PloS one |
Medium |
28982183
|
| 2019 |
NEK1 regulates retromer-mediated endosomal trafficking by phosphorylating VPS26B. NEK1 deficiency disrupts endosomal trafficking of plasma membrane proteins and leads to lysosomal degradation of A20 (a RIPK1 modulator), sensitizing cerebrovascular endothelial cells to RIPK1-dependent apoptosis and necroptosis, thus disrupting blood-brain barrier integrity. |
Phosphoproteomics, in vitro kinase assay (NEK1 phosphorylating VPS26B), NEK1 knockout mice, genetic inactivation of RIPK1, metabolic rescue studies, BBB permeability assays |
Nature communications |
High |
34376696
|
| 2020 |
NEK1 phosphorylates YAP1 at six residues in vitro, and NEK1 co-immunoprecipitates with YAP1. NEK1 overexpression leads to YAP1 accumulation (stabilization); NEK1 knockout leads to parallel decrease in YAP1 levels and reduced YAP-regulated gene expression, establishing a TLK1→NEK1→YAP1 axis. |
In vitro kinase assay (NEK1 phosphorylating YAP1 at 6 sites), co-immunoprecipitation, NEK1 knockout in NT1 cells, gene expression analysis |
Cancers |
Medium |
33297404
|
| 2020 |
TLK1-mediated phosphorylation of Nek1-T141 promotes VDAC1 phosphorylation and mitochondrial integrity. Cells overexpressing the NEK1-T141A phospho-deficient mutant show exaggerated apoptosis, altered mitochondrial oxygen consumption, increased glycolysis dependence, and cytochrome C leakage after doxorubicin treatment, placing TLK1 upstream of the Nek1-VDAC1 axis in apoptosis regulation. |
NEK1-T141A overexpression, mitochondrial respiration assays, cytochrome C fractionation, flow cytometry (sub-G1), western blot |
Cell cycle (Georgetown, Tex.) |
Medium |
31914854
|
| 2020 |
FBXO3 (substrate receptor of an SCF E3 ubiquitin ligase) binds and ubiquitylates C21ORF2 for proteasomal degradation. NEK1-mediated phosphorylation of C21ORF2 stabilizes it by reducing its interaction with FBXO3. C21ORF2 in turn stabilizes NEK1. The ALS-associated C21ORF2-V58L mutant is hyperphosphorylated by NEK1, escapes FBXO3-mediated degradation, and accumulates with NEK1. |
Co-immunoprecipitation, ubiquitination assay, in vitro kinase assay, protein stability assay, mutant analysis |
iScience |
Medium |
32891887
|
| 2023 |
NEK1 phosphorylates α-tubulin and importin-β1 in vitro. NEK1 loss-of-function (knockdown, kinase inhibition, patient mutation) causes disruptions in microtubule homeostasis and nuclear import in iPSC-derived motor neurons. Microtubule stabilization (with two distinct drug classes) restores both microtubule and nuclear import deficits, identifying NEK1 as a regulator of both pathways. |
Mass spectrometry interactome analysis, in vitro kinase assay (α-tubulin, importin-β1), iPSC-MN models (knockdown, kinase inhibitor, patient mutation), microtubule and nuclear import functional assays, pharmacological rescue |
Science advances |
High |
37585529
|
| 2023 |
Endogenous NEK1 and C21ORF2 form a tight complex in human cells. A C21ORF2 interaction domain (CID) at the NEK1 C-terminus is necessary for the association, and pathogenic mutations in this region disrupt the complex. NEK1 mutations that inhibit kinase activity or weaken C21ORF2 association severely compromise ciliogenesis, and C21ORF2 (like NEK1) is required for homologous recombination. |
Co-immunoprecipitation of endogenous proteins, AlphaFold structural modeling, domain mapping with pathogenic mutants, ciliogenesis assay, HR assay |
Life science alliance |
High |
37188479
|
| 2023 |
NEK1 phosphorylates YAP1 at Y407, and this phosphorylation promotes YAP1 nuclear retention and association with transcriptional co-activators AR and TEAD1. Pharmacological inhibition of TLK1 (J54) reverses Y407 phosphorylation and promotes cytoplasmic degradation of YAP1, suppressing androgen-independent transcriptional reprogramming. |
In vitro kinase assay (Y407 site identification), phospho-specific antibody, ChIP assay, GFP-YAP mutant (Y407F) transcriptome analysis, pharmacological inhibitor (J54) |
Biomedicines |
Medium |
36979713
|
| 2024 |
NEK1 loss-of-function ALS variants cause primary ciliary abnormality, cell cycle re-entry, and disrupted tubulin acetylation via AurA-HDAC6 pathway activation triggered by intracellular calcium overload. HDAC6 inhibition or Ca2+ chelation (BAPTA) rescues these deficits in patient fibroblasts and iPSC-MNs. |
Patient fibroblast and iPSC-MN studies, primary cilia immunostaining, calcium imaging, HDAC6 activity assay, tubulin acetylation western blot, pharmacological rescue |
Molecular neurodegeneration |
Medium |
40389989
|
| 2025 |
A novel NEK1 intronic variant causing SRPS eliminates the C-terminal 90 residues of NEK1 following the last coiled-coil region. This C-terminal fragment mediates interaction with VDAC1, and its absence is predicted to abolish VDAC1 phosphorylation by NEK1, linking NEK1 C-terminal truncation to ciliogenesis defects via disrupted VDAC1 regulation. |
Whole-exome sequencing, minigene splicing assay, bioinformatic protein interaction prediction, domain mapping of NEK1-VDAC1 interaction |
Bone |
Low |
40147672
|
| 2025 |
PML (promyelocytic leukemia protein) promotes SUMO-facilitated degradation of truncated/mutant Nek1t protein through PML nuclear bodies. In mice, PML ablation worsens ALS in heterozygous Nek1wt/t animals, and PML induction (via interferon pathway activation with poly(I:C)) clears Nek1t aggregates in alpha-motoneurons, dramatically reducing ALS symptoms and extending survival by 5 months. |
Nek1 Arg812Ter knock-in mice, Pml knockout cross, poly(I:C) induction, immunofluorescence of Nek1t aggregates, survival analysis, SUMO pathway analysis |
The FEBS journal |
High |
41804597
|