Affinage

PRKDC

DNA-dependent protein kinase catalytic subunit · UniProt P78527

Length
4128 aa
Mass
469.1 kDa
Annotated
2026-06-10
100 papers in source corpus 40 papers cited in narrative 40 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 6/6 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

PRKDC encodes DNA-PKcs, a nuclear PIKK-family serine/threonine kinase that serves as the catalytic engine of non-homologous end joining (NHEJ), the principal pathway for repairing DNA double-strand breaks (DSBs) (PMID:8041718, PMID:7855602). Recruited to free DNA ends by the Ku70/Ku80 heterodimer, DNA-PKcs assembles the catalytically active DNA-PK holoenzyme only in the presence of double-stranded DNA, with Ku and DNA coordinately inducing conformational changes that allosterically stimulate the kinase and protect ~30 bp of the DNA end within a binding tunnel (PMID:8041718, PMID:28840859, PMID:28652322, PMID:33385326). Activation is governed by autophosphorylation: hairpinned versus open DNA ends differentially trigger phosphorylation of the ABCDE (T2609) cluster, driving a structural rearrangement that widens the DNA-binding groove, destabilizes end binding to promote NHEJ progression, and licenses recruitment of the Artemis nuclease, which DNA-PKcs activates to open hairpins and cleave single-to-double-strand transitions during V(D)J recombination (PMID:17438073, PMID:34936881, PMID:32457294, PMID:32716029, PMID:15936993, PMID:35150269). DNA-PKcs initiates the broader DNA damage response by phosphorylating chromatin factors H2AX and KAP1 and promoting chromatin decondensation at breaks, and it shapes repair-pathway choice by negatively regulating ATM through direct phosphorylation and by suppressing hyper-recombination (PMID:31396623, PMID:27939942, PMID:19535303); catalytically inactive DNA-PKcs persists at lesions and causes greater genomic instability than protein loss, revealing distinct structural and catalytic functions (PMID:32015826, PMID:30072430). Loss of DNA-PKcs underlies the murine SCID immunodeficiency phenotype (PMID:7855601, PMID:7855602). Beyond canonical repair, DNA-PKcs has KU-directed roles in 18S rRNA processing within the small-subunit processome via U3 snRNA-driven assembly and T2609 phosphorylation (PMID:32103174), acts as a cytosolic DNA sensor driving STING-independent antiviral responses while phosphorylating and suppressing cGAS (PMID:31980485, PMID:33273464), and executes signaling functions at the Golgi (GOLPH3-MYO18A-mediated dispersal after damage) (PMID:24485452), mitochondria (Fis1-driven fragmentation and an ANT2/VDAC2 oxidative-phosphorylation complex) (PMID:35290083, PMID:36727301), telomeres (PMID:12426399, PMID:19244120, PMID:37653239), and replication forks (promoting fork reversal independent of NHEJ) (PMID:36130596).

Mechanistic history

Synthesis pass · year-by-year structured walk · 14 steps
  1. 1994 High

    Established that DNA-PK is an assembled holoenzyme requiring Ku and double-stranded DNA, defining the order of complex formation that underlies break recognition.

    Evidence Anti-Ku immunoprecipitation and reconstitution with purified components plus catalytic activity assay

    PMID:8041718

    Open questions at the time
    • Did not resolve the structural basis of activation
    • Did not identify physiological substrates beyond the complex itself
  2. 1995 High

    Linked DNA-PKcs genetically to immunodeficiency and DSB repair, showing the gene is responsible for the murine SCID defect and required for repair in human cells.

    Evidence Chromosomal complementation, co-localization mapping, and immunoblot/repair assays in SCID and M059J cells

    PMID:7855601 PMID:7855602

    Open questions at the time
    • Did not define the molecular step of NHEJ requiring DNA-PKcs
    • Catalytic versus structural contribution unresolved
  3. 2005 High

    Defined a downstream effector function by showing the Artemis:DNA-PKcs complex is a versatile endonuclease cleaving single-to-double-strand transitions, explaining hairpin/overhang processing.

    Evidence In vitro endonuclease assays with purified Artemis:DNA-PKcs on defined substrates

    PMID:15936993

    Open questions at the time
    • Did not define how phosphorylation gates the activity in cells
    • Structural arrangement of Artemis on DNA-PKcs not resolved
  4. 2007 High

    Resolved how autophosphorylation regulates NHEJ by showing it controls stability of DNA-PKcs on DNA ends rather than initial recruitment.

    Evidence Laser microirradiation and FRAP with kinase-dead and phospho-mutant GFP-DNA-PKcs in live cells

    PMID:17438073

    Open questions at the time
    • Did not identify the conformational basis of destabilization
    • Did not connect retention to specific pathway outcomes
  5. 2010 High

    Mapped DNA-PKcs into pathway-choice control by showing ABCDE autophosphorylation regulates repair-factor access and that DNA-PKcs and ATM coordinately govern NHEJ versus HR.

    Evidence I-SceI HR assays, epistasis with kinase-dead/null mutants, RAD51 foci, plus in vitro Artemis ssDNA endonuclease stimulation

    PMID:19535303 PMID:20117966

    Open questions at the time
    • Direct ATM phosphorylation of DNA-PKcs not yet defined
    • Site-level basis of pathway switching incomplete
  6. 2014 High

    Extended DNA-PK signaling beyond the nucleus by showing damage-induced phosphorylation of GOLPH3 drives MYO18A-dependent Golgi dispersal affecting survival.

    Evidence siRNA knockdown, phospho-specific antibodies, reciprocal co-IP, Golgi imaging, and survival assays

    PMID:24485452

    Open questions at the time
    • Phosphorylation sites on GOLPH3 not mapped here
    • Connection to nuclear DSB sensing unclear
  7. 2016 High

    Defined the molecular basis of DNA-PKcs/ATM crosstalk by showing DNA-PKcs directly phosphorylates and inhibits ATM, establishing reciprocal kinase regulation.

    Evidence In vitro kinase assays with purified proteins, ATM site mutagenesis, and DNA-PKcs KO/inhibition signaling assays

    PMID:27939942

    Open questions at the time
    • In vivo stoichiometry of ATM inhibition unresolved
    • Temporal regulation across DDR phases unclear
  8. 2017 High

    Provided the first cryo-EM structures of DNA-PKcs and the holoenzyme, revealing the α-solenoid architecture, the DNA-binding tunnel protecting end DNA, and allosteric stimulation by Ku/DNA.

    Evidence Cryo-EM (4.4–6.6 Å) of DNA-PKcs and DNA-PK holoenzyme with activity assays

    PMID:28652322 PMID:28840859

    Open questions at the time
    • Activated-state conformational changes not yet captured
    • Autophosphorylation-driven rearrangements not visualized
  9. 2019 High

    Established DNA-PKcs kinase activity as initiator of the DDR by showing it phosphorylates H2AX and KAP1 and drives chromatin decondensation and DDR factor recruitment at breaks.

    Evidence Kinase-domain-inactivated human cells, irradiation, phospho-readouts, and recruitment kinetics; PARylation by PARP1 regulating chromatin retention

    PMID:31396623 PMID:31485633

    Open questions at the time
    • Quantitative hierarchy with ATM-driven DDR signaling unresolved
    • PARylation site mapping incomplete (Medium-confidence)
  10. 2020 High

    Uncovered non-canonical RNA-, immune-, and metabolism-linked functions: KU-directed assembly on U3 snRNA for 18S rRNA processing, DNA-PK as a STING-independent DNA sensor, and cGAS phosphorylation suppressing innate immunity.

    Evidence Mouse knockin models, U3 snRNA activation of purified DNA-PK, SSU processome co-purification, phosphoproteomics (HSPA8), in vitro cGAS kinase assay, patient-derived cells

    PMID:31980485 PMID:32103174 PMID:33273464

    Open questions at the time
    • Mechanistic separation of nuclear repair versus nucleolar/immune roles incomplete
    • How RNA versus DNA selects functional output unclear
  11. 2021 High

    Revealed the autophosphorylation-driven functional switch structurally: open versus hairpin ends differentially trigger ABCDE phosphorylation, widening the DNA groove for Artemis recruitment and hairpin cleavage, with Artemis locking the kinase inactive.

    Evidence Cryo-EM of DNA-PK pre/post-autophosphorylation and with Artemis/hairpin, plus kinase and nuclease assays; additional substrate-specific activation by hairpin ends

    PMID:32716029 PMID:34936881

    Open questions at the time
    • Order of events in the full synaptic complex not fully defined
    • Substrate selection rules beyond Artemis incomplete
  12. 2021 High

    Demonstrated a substrate-stabilizing role in cancer stem cells: DNA-PKcs phosphorylates SOX2 at S251 to block WWP2-mediated degradation, maintaining glioma stem cells.

    Evidence Mass spectrometry, co-IP, S251 mutagenesis, ubiquitination assays, and xenograft DNA-PKcs inhibition

    PMID:34193614

    Open questions at the time
    • Whether nuclear DNA-PK pool serves this function during damage unclear
    • Generality across other transcription factors untested
  13. 2022 High

    Defined mitochondrial and conformational dimensions: DNA-PKcs phosphorylates Fis1-T34 to drive fragmentation, structural studies mapped the Artemis/XRCC4 mutually exclusive interface, and dimeric synaptic conformations captured NHEJ transition states.

    Evidence In vitro kinase assays and Fis1-T34A knockin mice; cryo-EM of basal Artemis:DNA-PKcs and dimeric complexes; structures with ATPγS and inhibitors; V(D)J domain-mapping mutagenesis; fork-reversal EM

    PMID:34987222 PMID:35150269 PMID:35290083 PMID:35801871 PMID:36130596 PMID:37256947

    Open questions at the time
    • How cytoplasmic versus nuclear pools are partitioned unresolved
    • Coupling of fork-reversal role to canonical kinase signaling unclear
  14. 2023 High

    Extended DNA-PKcs into mitochondrial energetics and telomere protection: it forms the ANT2/VDAC2 (DAV) complex sustaining ADP-ATP exchange, dissociated by ATM-mediated T2609 phosphorylation, and cooperates with TRF2 to repress MRN-initiated resection at blunt telomeres.

    Evidence Co-IP, mitochondrial fractionation, Seahorse/membrane-potential assays, ATM kinase assays; in vitro/in vivo resection assays with AlphaFold-Multimer prediction

    PMID:36727301 PMID:37653239

    Open questions at the time
    • Structural basis of DAV complex assembly not resolved
    • How a nuclear kinase is partitioned to mitochondria mechanistically unclear

Open questions

Synthesis pass · forward-looking unresolved questions
  • How DNA-PKcs is spatially and functionally partitioned among its nuclear repair, nucleolar rRNA-processing, cytosolic sensing, Golgi, mitochondrial, and replication-fork roles—and what signals route the same kinase to each—remains unresolved.
  • No unifying model of subcellular trafficking
  • Substrate selection rules across compartments undefined
  • Relative physiological importance of non-canonical roles unquantified

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 6 GO:0016740 transferase activity 4 GO:0003677 DNA binding 3 GO:0003723 RNA binding 1 GO:0140657 ATP-dependent activity 1
Localization
GO:0005634 nucleus 2 GO:0005739 mitochondrion 2 GO:0005829 cytosol 2 GO:0005730 nucleolus 1 GO:0005794 Golgi apparatus 1
Pathway
R-HSA-168256 Immune System 4 R-HSA-73894 DNA Repair 4 R-HSA-1266738 Developmental Biology 2 R-HSA-1430728 Metabolism 1 R-HSA-69306 DNA Replication 1 R-HSA-8953854 Metabolism of RNA 1
Partners
ATMCGASDCLRE1C (ARTEMIS)FIS1GOLPH3SOX2XRCC5 (KU80)XRCC6 (KU70)
Complex memberships
Artemis:DNA-PKcs complexDAV complex (DNA-PKcs–ANT2–VDAC2)DNA-PK holoenzyme (DNA-PKcs–Ku70/Ku80)small-subunit (SSU) processome

Evidence

Reading pass · 40 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1994 The DNA-PK holoenzyme (Ku protein + p350/DNA-PKcs) assembles on double-stranded DNA: Ku binds first to free DNA ends, then recruits p350 to form a catalytically active complex. Reconstitution experiments showed the complex forms only in the presence of double-stranded DNA. Immunoprecipitation with anti-Ku antibodies, reconstitution with purified components, catalytic activity assay Proceedings of the National Academy of Sciences of the United States of America High 8041718
1992 DNA-PKcs is a nuclear serine/threonine protein kinase activated by direct interaction with DNA (not using DNA as template/substrate) and phosphorylates DNA-binding proteins including transcription factors, suggesting a role in modulating transcriptional activity. Biochemical kinase assays, nuclear fractionation Critical reviews in eukaryotic gene expression Medium 1486241
1995 DNA-PKcs (p350) is the gene responsible for the murine SCID defect: p350 and the SCID-complementing gene co-localize to human chromosome 8q11, chromosomal fragments expressing p350 complement the SCID phenotype, and p350 protein is greatly reduced in SCID mouse cells. Chromosomal complementation, co-localization mapping, western blot of SCID-derived cells Science (New York, N.Y.) High 7855601 7855602
1995 Absence of the p350 subunit of DNA-PK in the radiosensitive human glioma cell line M059J is associated with defective DNA double-strand break repair, demonstrating that DNA-PK kinase activity is required for DSB repair. Immunoblot for p350 expression, DNA-PK kinase activity assay, DSB repair assay in human cell lines Science (New York, N.Y.) High 7855602
2005 The Artemis:DNA-PKcs complex cleaves a wide range of DNA substrates containing single-to-double-strand transitions (heterologous loops, stem-loops, flaps, gapped substrates) near the transition region; this versatile endonuclease activity is activated by DNA-PKcs binding and phosphorylation of Artemis. In vitro endonuclease assays with purified Artemis:DNA-PKcs complex on defined DNA substrates DNA repair High 15936993
2007 DNA-PKcs accumulates at DSB sites in a Ku80-dependent manner. Kinase activity and autophosphorylation status do not affect initial recruitment but do regulate the stability of DNA-PKcs binding to DNA ends; impaired autophosphorylation leads to prolonged retention at unrepaired DSBs, suggesting autophosphorylation facilitates NHEJ by destabilizing the interaction with DNA ends. Laser-induced DSB formation in live cells, FRAP (fluorescence recovery after photobleaching), live-cell imaging of GFP-DNA-PKcs The Journal of cell biology High 17438073
2010 DNA-PKcs autophosphorylation at the ABCDE cluster regulates access of repair factors to DNA ends. Kinase-dead DNA-PKcs (DNA-PKcs-KR) cells show hyperrecombination 2-3 fold above DNA-PKcs null, and ATM-dependent phosphorylation of DNA-PKcs-KR contributes to this hyperrecombination phenotype; DNA-PKcs and ATM coordinately regulate the NHEJ/HR pathway choice. Direct repeat HR assay with I-SceI nuclease, epistasis analysis with kinase-dead and null mutants, RAD51 foci analysis DNA repair High 19535303
2010 DNA-PKcs regulates Artemis single-stranded DNA endonuclease activity: purified Artemis has intrinsic ssDNA endonuclease activity that is stimulated by DNA-PKcs; the divalent cation and sequence dependence matches that of the Artemis:DNA-PKcs double-stranded endonuclease activity. In vitro endonuclease assay with highly purified Artemis, addition of DNA-PKcs, antibody modulation DNA repair High 20117966
2010 DNA-PKcs selectively stimulates WRN helicase activity (but not WRN exonuclease) on telomere D-loop model substrates in vitro, and DNA-PKcs knockdown reduces telomeric G-tail length in vivo; this effect is reversed by WRN helicase overexpression, indicating cooperative roles of DNA-PKcs and WRN in maintaining telomeric G-tails. In vitro helicase/exonuclease assay with purified DNA-PKcs and WRN, siRNA knockdown, telomere G-tail length measurement Aging Medium 20519774
2014 DNA damage triggers DNA-PK-dependent phosphorylation of GOLPH3, which increases GOLPH3 interaction with MYO18A, applying tensile force to the Golgi and resulting in Golgi dispersal throughout the cytoplasm. Depletion of DNA-PK, GOLPH3, or MYO18A reduces survival after DNA damage. siRNA knockdown, phospho-specific antibodies, co-immunoprecipitation, fluorescence microscopy of Golgi morphology, cell survival assays Cell High 24485452
2016 DNA-PKcs phosphorylates ATM at specific sites, inhibiting ATM activity. Chemical inhibition or genetic deletion of DNA-PKcs leads to hyperactive ATM; pre-incubation of ATM with active DNA-PKcs reduces ATM activity in vitro. Phospho-mimetic mutations at DNA-PKcs target sites in ATM inhibit ATM activity and impair ATM signaling after DNA damage. In vitro kinase assay with purified proteins, mutagenesis of ATM phosphorylation sites, DNA damage signaling assays in human cells with DNA-PKcs KO/inhibition Molecular cell High 27939942
2002 DNA-PKcs functionally interacts with telomerase in telomere length maintenance: Terc(-/-)/DNA-PKcs(-/-) double-knockout mice show accelerated telomere shortening compared to Terc(-/-) alone. DNA-PKcs is also required for end-to-end chromosome fusions and apoptosis triggered by critically short telomeres. Double-knockout mouse model, telomere length analysis (FISH/TRF), cytogenetic analysis The EMBO journal High 12426399
2009 Phosphorylation of DNA-PKcs at the Thr-2609 cluster is a critical event for proper telomere end-processing; DNA-PKcs deficiency leads to uncapped telomeres that are inappropriately processed as DSBs, participating in spontaneous and radiation-induced telomere-DSB fusions requiring ligase IV. Cytogenetic analysis in DNA-PKcs-deficient mouse and human cells, phospho-specific immunoblot, irradiation experiments Cancer research Medium 19244120
2017 Cryo-EM structure of the human DNA-PK holoenzyme shows DNA-PKcs, KU70, KU80, and DNA forming a 650-kDa heterotetramer; DNA-PKcs N-terminal α-solenoid adopts a double-ring fold; together with KU70/80, DNA-PKcs forms a DNA-binding tunnel that protects ~30-bp DNA end. KU70/80 and DNA coordinately induce conformational changes in DNA-PKcs and allosterically stimulate its kinase activity. Cryo-electron microscopy (6.6 Å resolution), biochemical activity assays Cell research High 28652322 28840859
2017 Cryo-EM structures of human DNA-PKcs (4.4 Å) and DNA-PK holoenzyme (5.8 Å) reveal that DNA-PKcs has three segments: N-terminal region (arm and bridge), circular cradle, and head with kinase domain; the C-terminal globular domain of Ku80 interacts with the arm of DNA-PKcs. Cryo-electron microscopy structural analysis Proceedings of the National Academy of Sciences of the United States of America High 28652322
2020 Cryo-EM structures of DNA-PKcs bound to DNA end and in complex with Ku70/80-DNA in inactive and activated states (3.7 Å overall; 3.2 Å FATKIN) reveal that kinase activation involves stretching and twisting within solenoid segments, loosening DNA-end binding. This structural plasticity of HEAT repeats represents a regulatory mechanism for PIKK family kinases. Cryo-electron microscopy, structural comparison of inactive vs. activated states Molecular cell High 33385326
2020 DNA-PKcs has a KU-dependent role in rRNA processing and haematopoiesis: KU drives assembly of DNA-PKcs on cellular RNAs including U3 snRNA; U3 snRNA activates purified DNA-PK and triggers DNA-PKcs phosphorylation at T2609. DNA-PK resides in nucleoli in an rRNA-dependent manner and co-purifies with the small subunit processome. Blocking T2609 cluster phosphorylation (but not S2056 cluster) causes defects in 18S rRNA processing and bone marrow failure. Mouse knockin models (kinase-dead, T2609A), purified protein activation assays with U3 snRNA, nucleolar fractionation, co-purification with SSU processome, ribosome profiling Nature High 32103174
2020 Human DNA-PK functions as a sensor in a STING-independent DNA sensing pathway (SIDSP): DNA-PK drives a broad antiviral response; HSPA8/HSC70 is identified as a target of inducible phosphorylation downstream of DNA-PK in this pathway. Viral proteins E1A (adenovirus) and ICP0 (HSV-1) block this response. Genetic deletion and inhibition of DNA-PK, phosphoproteomics identifying HSPA8/HSC70 as substrate, viral infection assays, antiviral response measurement Science immunology High 31980485
2020 DNA-PK phosphorylates cGAS and suppresses its enzymatic activity. DNA-PK deficiency reduces cGAS phosphorylation and promotes antiviral innate immune responses. Cells from DNA-PKcs-deficient mice or patients with PRKDC missense mutations exhibit an inflammatory gene expression signature. In vitro kinase assay with DNA-PKcs and cGAS, co-immunoprecipitation, genetic mouse models, patient-derived cells Nature communications High 33273464
2021 Cryo-EM structures of DNA-PK bound to DNA ends before and after autophosphorylation, and in complex with Artemis and a DNA hairpin, reveal a functional switch: open DNA ends inhibit cis-autophosphorylation of the ABCDE cluster but activate phosphorylation of other targets; hairpin ends promote ABCDE cis-autophosphorylation. Phosphorylation of four Thr residues in ABCDE causes gross structural rearrangement widening the DNA-binding groove for Artemis recruitment and hairpin cleavage. Artemis locks DNA-PK in a kinase-inactive state. Cryo-EM structural analysis of multiple states, in vitro kinase and nuclease assays Molecular cell High 34936881
2019 DNA-PKcs kinase activity is required for initiation of the DDR immediately after DSB induction: it drives phosphorylation of chromatin factors H2AX and KAP1, promotes local chromatin decondensation near DSB sites, and facilitates recruitment of DDR machinery. Loss of DNA-PKcs kinase activity markedly decreases DDR factor recruitment to DSBs. Kinase-domain inactivating human cell line, ionizing radiation, γH2AX and KAP1 phosphorylation assays, chromatin decondensation measurement, recruitment kinetics of DDR factors Nucleic acids research High 31396623
2020 DNA-PKcs is neddylated at its kinase domain by the E2-conjugating enzyme UBE2M and E3 ligase HUWE1; inhibition of HUWE1-dependent neddylation impairs DNA-PKcs autophosphorylation at Ser2056 and reduces NHEJ efficiency. Immunoprecipitation, co-immunoprecipitation, HUWE1/UBE2M knockdown, NHEJ reporter assay, phospho-Ser2056 immunoblot Cell death & disease Medium 32457294
2022 Cryo-EM structural analysis of the basal (pre-activated) Artemis:DNA-PKcs complex shows the Artemis catalytic domain is positioned externally to DNA-PKcs prior to ABCDE autophosphorylation; both Artemis catalytic and regulatory domains interact with the N-HEAT and FAT domains of DNA-PKcs. A mutually exclusive binding site for Artemis and XRCC4 on DNA-PKcs was defined, and an XRCC4 peptide disrupts the Artemis:DNA-PKcs complex. Cryo-EM structural analysis, agarose-acrylamide gel complex stabilization, peptide competition assay Nucleic acids research High 35801871
2022 Cryo-EM structures of DNA-PKcs in three distinct dimeric conformations represent transition states during NHEJ: upon autophosphorylation, the long-range synaptic complex undergoes conformational change with both Ku and DNA-PKcs rotating outward to promote DNA break exposure and DNA-PKcs dissociation. Single-particle cryo-electron microscopy of NHEJ complexes at different stages Science advances High 37256947
2022 Cryo-EM structures of human DNA-PKcs with ATPγS and four inhibitors (wortmannin, NU7441, AZD7648, M3814) reveal the ATP binding mode and show that inhibitor binding causes movement of the PIKK regulatory domain (PRD), revealing a connection between the p-loop and PRD conformations. Inhibitors function through direct ATP competition and do not negatively allosterically affect holoenzyme assembly. Cryo-EM structural analysis, electrophoretic mobility shift assay (EMSA) for holoenzyme assembly Nature High 34987222
2022 DNA-PKcs promotes fork reversal at stressed replication forks in a manner independent of its NHEJ role; cells lacking DNA-PKcs activity show increased DNA damage during S-phase and sensitivity to replication stress. Prevention of fork reversal by DNA-PKcs inhibition restores chemotherapy sensitivity in BRCA2-deficient tumors with acquired PARP inhibitor resistance. Electron microscopy of replication intermediates, DNA fiber assay, DNA-PKcs inhibitor/knockout, BRCA2-deficient tumor model Molecular cell High 36130596
2022 DNA-PKcs interacts with and phosphorylates Fis1 at Thr34 in its TQ motif, increasing Fis1 affinity for Drp1 and inducing mitochondrial fragmentation; knockin mice with non-phosphorylatable T34A Fis1 show improved renal function and reduced mitochondrial fragmentation in acute kidney injury. Cytoplasmic localization of DNA-PKcs was detected in injured kidney tissues. Co-immunoprecipitation, in vitro kinase assay, Fis1-T34A knockin mice, mitochondrial morphology analysis, human patient urinary sediment analysis Science signaling High 35290083
2022 Physical ARTEMIS:DNA-PKcs interaction is necessary for V(D)J recombination: the L3062R pathogenic mutation in DNA-PKcs impairs physical interaction with Artemis; specific mutations in Artemis (in two conserved regions) that disrupt interaction with DNA-PKcs impair V(D)J recombination. Minimal interaction fragments were mapped: 42 aa from FAT region 2 of DNA-PKcs (PKcs3041-3082) and 26 aa from Artemis (ARM378-403). Mutagenesis, co-immunoprecipitation, V(D)J recombination assay, domain mapping with minimal fragments Nucleic acids research High 35150269
2023 DNA-PKcs directly interacts with mitochondrial proteins ANT2 and VDAC2 forming the DAV complex, which supports ADP-ATP exchange across mitochondrial membranes to sustain oxidative phosphorylation and membrane potential. The DAV complex dissociates in response to oxidative stress, attenuating ADP-ATP exchange; dissociation is mediated by ATM-dependent phosphorylation of DNA-PKcs at the Thr2609 cluster. Co-immunoprecipitation, mitochondrial fractionation, membrane potential assays, Seahorse metabolic analysis, DNA-PKcs-deficient cell lines, ATM kinase assay The EMBO journal High 36727301
2023 DNA-PK and TRF2 cooperate to repress MRN-initiated resection at leading-end (blunt) telomeres: DNA-PK represses MRN-dependent long-range resection, while the iDDR of TRF2 inhibits MRN-CtIP endonuclease activity that would otherwise cleave DNA-PK off blunt telomere ends. AlphaFold-Multimer predicts conserved iDDR association with Rad50, potentially interfering with CtIP binding. In vitro resection assays, in vivo telomere resection analysis, AlphaFold-Multimer structural prediction with experimental validation Nature structural & molecular biology High 37653239
2021 DNA-PKcs (DNA-PK catalytic subunit) phosphorylates SOX2 at S251, stabilizing SOX2 by preventing WWP2-mediated ubiquitination and promoting glioma stem cell maintenance. Upon DNA damage, the DNA-PK complex dissociates from SOX2, allowing WWP2 interaction and SOX2 degradation, triggering differentiation. Mass spectrometry of SOX2-binding proteins, co-immunoprecipitation, site-directed mutagenesis of S251, ubiquitination assays, in vitro and in vivo (xenograft) DNA-PKcs inhibition Science translational medicine High 34193614
2020 DNA-PKcs kinase activity and autophosphorylation regulate kinase complex conformation and dissociation during NHEJ; expression of catalytically inactive DNA-PKcs causes more genomic instability than loss of the protein itself (structural function), because kinase-dead DNA-PKcs persists at DNA lesions and alters repair pathway choice. Mouse models expressing kinase-dead DNA-PKcs, genomic instability assays, comparison to protein-null models Cell & bioscience Medium 32015826
2019 DNA-PARylation of DNA-PKcs by PARP1 regulates DNA-PK activity: DNA-PKcs is PARylated after DNA damage, PARP inhibition (olaparib) prevents DNA-PKcs detachment from chromatin and maintains DNA-PKcs Ser2056 autophosphorylation; olaparib and DNA-PK inhibition synergize to suppress cell survival. Immunoprecipitation, immunofluorescence, chromatin fractionation, phospho-Ser2056 immunoblot, cell survival assays Molecular medicine reports Medium 31485633
2020 Activation of DNA-PK by hairpinned DNA ends is substrate-specific: hairpinned DNA ends do not activate DNA-PK toward p53, XRCC4, XLF, or HSP90, but robustly stimulate ABCDE cluster autophosphorylation, which is required for Artemis activation. This reveals a multi-step mechanism of kinase activation. In vitro kinase assays with defined DNA substrates (hairpinned vs open ends), comparison across multiple substrates Nucleic acids research High 32716029
2021 The CRL4A-DTL ubiquitin ligase complex targets DNA-PKcs for nuclear proteasomal degradation; overexpression of CUL4A or DTL reduces NHEJ repair efficiency and increases DSB accumulation, leading to genomic instability and malignant transformation. Co-immunoprecipitation, ubiquitination assay, NHEJ reporter assay, γH2AX measurement, overexpression in normal cells Oncogene Medium 33627782
2024 PRKDC recruits GDE2 to enhance stability of GNAS protein, which activates AKT phosphorylation, conferring doxorubicin resistance in osteosarcoma. The PRKDC-GDE2-GNAS-AKT regulatory axis was identified by a kinome-wide CRISPR screen. CRISPR kinome screen, co-immunoprecipitation of PRKDC and GDE2, GNAS stability assays, AKT phosphorylation analysis, xenograft and organoid models Cancer research Medium 38900943
2018 DNA-PKcs has a kinase-dependent function in suppressing microhomology-mediated end joining (MMEJ) during class switch recombination (CSR) and a structural (kinase-independent) role in orientation of CSR; kinase-dead DNA-PKcs severely compromises CSR to IgG1 while DNA-PKcs deletion does not, revealing distinct structural and catalytic roles. Mouse B cell models with kinase-dead vs. null DNA-PKcs, high-throughput sequencing of CSR junctions, translocation analysis Proceedings of the National Academy of Sciences of the United States of America High 30072430
2001 BCR-ABL down-regulates DNA-PKcs via proteasome-dependent degradation that requires BCR-ABL tyrosine kinase activity, resulting in marked DNA repair deficiency and increased sensitivity to ionizing radiation. Stable and inducible BCR-ABL expression in hematopoietic cells, proteasome inhibitor experiments, western blot, irradiation sensitivity assays Blood Medium 11264175
2012 Upon ionizing radiation, nuclear EGFR associates with DNA-PK, which phosphorylates PNPase (polynucleotide phosphorylase) at Ser-776; phospho-mimetic S776D PNPase has impaired ribonuclease activity, while the non-phosphorylatable S776A mutant retains ribonuclease activity and degrades c-MYC mRNA, affecting radioresistance. Co-immunoprecipitation, site-directed mutagenesis of PNPase, in vitro ribonuclease assays, knockdown experiments The Journal of biological chemistry High 22815474
2020 DNA-PKcs promotes cardiac ischemia-reperfusion injury through direct interaction with BI-1 (Bax inhibitor-1), promoting BI-1 degradation without affecting BI-1 transcription. Loss of DNA-PKcs stabilizes BI-1, protecting mitochondria; concurrent BI-1 knockout abrogates the cardioprotection of DNA-PKcs deletion. Cardiomyocyte-specific DNA-PKcs knockout mice, co-immunoprecipitation, double-knockout epistasis, mitochondrial function assays Basic research in cardiology Medium 31919590

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1995 DNA-dependent kinase (p350) as a candidate gene for the murine SCID defect. Science (New York, N.Y.) 590 7855601
1995 Absence of p350 subunit of DNA-activated protein kinase from a radiosensitive human cell line. Science (New York, N.Y.) 507 7855602
2001 DNA-PK, ATM and ATR as sensors of DNA damage: variations on a theme? Current opinion in cell biology 420 11248557
2005 The life and death of DNA-PK. Oncogene 344 15592499
2007 Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks. The Journal of cell biology 335 17438073
2014 DNA-PK: a dynamic enzyme in a versatile DSB repair pathway. DNA repair 271 24680878
2003 ATM, ATR and DNA-PK: initiators of the cellular genotoxic stress responses. Carcinogenesis 228 12919958
2014 Beyond DNA repair: DNA-PK function in cancer. Cancer discovery 209 25168287
2014 DNA damage triggers Golgi dispersal via DNA-PK and GOLPH3. Cell 203 24485452
2008 DNA-PK: the means to justify the ends? Advances in immunology 200 19117531
2010 Targeting DNA-PKcs and ATM with miR-101 sensitizes tumors to radiation. PloS one 183 20617180
2010 A structural model for regulation of NHEJ by DNA-PKcs autophosphorylation. DNA repair 175 21030321
2020 ATM, ATR and DNA-PKcs kinases-the lessons from the mouse models: inhibition ≠ deletion. Cell & bioscience 174 32015826
1994 DNA-dependent protein kinase (Ku protein-p350 complex) assembles on double-stranded DNA. Proceedings of the National Academy of Sciences of the United States of America 166 8041718
1992 The nuclear serine/threonine protein kinase DNA-PK. Critical reviews in eukaryotic gene expression 163 1486241
2020 Human DNA-PK activates a STING-independent DNA sensing pathway. Science immunology 148 31980485
2019 DNA-PK as an Emerging Therapeutic Target in Cancer. Frontiers in oncology 142 31380275
2020 DNA-PKcs: A Multi-Faceted Player in DNA Damage Response. Frontiers in genetics 139 33424929
2005 The Artemis:DNA-PKcs endonuclease cleaves DNA loops, flaps, and gaps. DNA repair 135 15936993
2020 DNA-PKcs promotes cardiac ischemia reperfusion injury through mitigating BI-1-governed mitochondrial homeostasis. Basic research in cardiology 130 31919590
2001 BCR-ABL down-regulates the DNA repair protein DNA-PKcs. Blood 128 11264175
2024 Quercetin inhibits necroptosis in cardiomyocytes after ischemia-reperfusion via DNA-PKcs-SIRT5-orchestrated mitochondrial quality control. Phytotherapy research : PTR 120 38447978
2009 DNA-PKcs and ATM co-regulate DNA double-strand break repair. DNA repair 112 19535303
2020 DNA-PK deficiency potentiates cGAS-mediated antiviral innate immunity. Nature communications 109 33273464
2020 Structure of an activated DNA-PK and its implications for NHEJ. Molecular cell 109 33385326
2016 Regulation of the DNA Damage Response by DNA-PKcs Inhibitory Phosphorylation of ATM. Molecular cell 109 27939942
2002 Functional interaction between DNA-PKcs and telomerase in telomere length maintenance. The EMBO journal 106 12426399
2022 DNA-PKcs promotes sepsis-induced multiple organ failure by triggering mitochondrial dysfunction. Journal of advanced research 97 36328752
2022 DNA-PKcs interacts with and phosphorylates Fis1 to induce mitochondrial fragmentation in tubular cells during acute kidney injury. Science signaling 95 35290083
2017 Cryo-EM structure of human DNA-PK holoenzyme. Cell research 95 28840859
2018 Synthetically Lethal Interactions of ATM, ATR, and DNA-PKcs. Trends in cancer 89 30352678
2023 Simultaneous inhibition of DNA-PK and Polϴ improves integration efficiency and precision of genome editing. Nature communications 88 37580318
2021 Inhibiting DNA-PK induces glioma stem cell differentiation and sensitizes glioblastoma to radiation in mice. Science translational medicine 80 34193614
2020 DNA-PKcs has KU-dependent function in rRNA processing and haematopoiesis. Nature 77 32103174
2019 DNA-PKcs promotes chromatin decondensation to facilitate initiation of the DNA damage response. Nucleic acids research 73 31396623
2021 Autophosphorylation transforms DNA-PK from protecting to processing DNA ends. Molecular cell 71 34936881
2009 Telomere dysfunction and DNA-PKcs deficiency: characterization and consequence. Cancer research 70 19244120
2017 Cryo-EM structure of the DNA-PK holoenzyme. Proceedings of the National Academy of Sciences of the United States of America 68 28652322
2012 DNA-PKcs expression predicts response to radiotherapy in prostate cancer. International journal of radiation oncology, biology, physics 66 22494583
2022 Structural insights into inhibitor regulation of the DNA repair protein DNA-PKcs. Nature 64 34987222
2020 Targeting DNA-PK in cancer. Mutation research 62 32172133
2016 Single-cell imaging of normal and malignant cell engraftment into optically clear prkdc-null SCID zebrafish. The Journal of experimental medicine 62 27810924
2020 Selective DNA-PKcs inhibition extends the therapeutic index of localized radiotherapy and chemotherapy. The Journal of clinical investigation 61 31581151
2017 DNA-PKcs, ATM, and ATR Interplay Maintains Genome Integrity during Neurogenesis. The Journal of neuroscience : the official journal of the Society for Neuroscience 60 28123024
2022 DNA-PKcs: A Targetable Protumorigenic Protein Kinase. Cancer research 58 34893509
2020 Uncovering DNA-PKcs ancient phylogeny, unique sequence motifs and insights for human disease. Progress in biophysics and molecular biology 54 33035590
2006 The DNA repair complex DNA-PK, a pharmacological target in cancer chemotherapy and radiotherapy. Pathologie-biologie 54 16563661
2009 DNA-PKcs deficiency in human: long predicted, finally found. Current opinion in allergy and clinical immunology 53 19823081
2009 Do DNA repair genes OGG1, XRCC3 and XRCC7 have an impact on susceptibility to bladder cancer in the North Indian population? Mutation research 52 19815090
2010 DNA-PKcs regulates a single-stranded DNA endonuclease activity of Artemis. DNA repair 50 20117966
2020 PRKDC: new biomarker and drug target for checkpoint blockade immunotherapy. Journal for immunotherapy of cancer 47 32238472
2012 Nuclear EGFR suppresses ribonuclease activity of polynucleotide phosphorylase through DNAPK-mediated phosphorylation at serine 776. The Journal of biological chemistry 46 22815474
2022 DNA-PKcs promotes fork reversal and chemoresistance. Molecular cell 44 36130596
2021 Role of PRKDC in cancer initiation, progression, and treatment. Cancer cell international 43 34702253
2014 Targeting DNA-PKcs and telomerase in brain tumour cells. Molecular cancer 43 25307264
2011 Unique and redundant functions of ATM and DNA-PKcs during V(D)J recombination. Cell cycle (Georgetown, Tex.) 41 21673501
2011 Irinotecan and DNA-PKcs inhibitors synergize in killing of colon cancer cells. Investigational new drugs 40 21221710
1996 p53 induction, cell cycle checkpoints, and apoptosis in DNAPK-deficient scid mice. Carcinogenesis 40 9006086
2015 DNA-PKcs deficiency inhibits glioblastoma cell-derived angiogenesis after ionizing radiation. Journal of cellular physiology 39 25294801
2008 Genetic variants of the XRCC7 gene involved in DNA repair and risk of human bladder cancer. International journal of urology : official journal of the Japanese Urological Association 39 18422577
2020 DNA-PK in human malignant disorders: Mechanisms and implications for pharmacological interventions. Pharmacology & therapeutics 36 32610116
2021 Structural insights into the role of DNA-PK as a master regulator in NHEJ. Genome instability & disease 35 34723130
2020 Beyond DNA Repair: DNA-PKcs in Tumor Metastasis, Metabolism and Immunity. Cancers 35 33207636
2005 DNA-PKcs-dependent signaling of DNA damage in Dictyostelium discoideum. Current biology : CB 32 16243037
2020 Discovery and development of novel DNA-PK inhibitors by targeting the unique Ku-DNA interaction. Nucleic acids research 31 33119767
2002 DNA-PKcs mutations in dogs and horses: allele frequency and association with neoplasia. Gene 31 11867233
2023 DNA-PK and the TRF2 iDDR inhibit MRN-initiated resection at leading-end telomeres. Nature structural & molecular biology 29 37653239
2020 Prevalence of PRKDC mutations and association with response to immune checkpoint inhibitors in solid tumors. Molecular oncology 29 32502294
2010 Polymorphic variants of DNA repair gene XRCC3 and XRCC7 and risk of prostate cancer: a study from North Indian population. DNA and cell biology 29 20590474
2010 Coordination of DNA-PK activation and nuclease processing of DNA termini in NHEJ. Antioxidants & redox signaling 29 20698792
2024 PRKDC Induces Chemoresistance in Osteosarcoma by Recruiting GDE2 to Stabilize GNAS and Activate AKT. Cancer research 28 38900943
2023 Cryo-EM visualization of DNA-PKcs structural intermediates in NHEJ. Science advances 28 37256947
2020 HUWE1-dependent DNA-PKcs neddylation modulates its autophosphorylation in DNA damage response. Cell death & disease 28 32457294
2019 DNA‑PKcs PARylation regulates DNA‑PK kinase activity in the DNA damage response. Molecular medicine reports 28 31485633
2022 Structural analysis of the basal state of the Artemis:DNA-PKcs complex. Nucleic acids research 27 35801871
2019 Pleiotropic Impact of DNA-PK in Cancer and Implications for Therapeutic Strategies. Clinical cancer research : an official journal of the American Association for Cancer Research 27 31266833
2018 The role of DNA-PK in aging and energy metabolism. The FEBS journal 27 29453899
2017 Small molecule inhibitors of DNA-PK for tumor sensitization to anticancer therapy. Journal of physiology and pharmacology : an official journal of the Polish Physiological Society 27 28820390
2016 DNA-PKcs: A promising therapeutic target in human hepatocellular carcinoma? DNA repair 27 27789167
2010 Cooperation of DNA-PKcs and WRN helicase in the maintenance of telomeric D-loops. Aging 27 20519774
2016 Restoration of ATM Expression in DNA-PKcs-Deficient Cells Inhibits Signal End Joining. Journal of immunology (Baltimore, Md. : 1950) 26 26921311
2021 LINC-PINT impedes DNA repair and enhances radiotherapeutic response by targeting DNA-PKcs in nasopharyngeal cancer. Cell death & disease 25 33963177
2019 Structural insights into the critical DNA damage sensors DNA-PKcs, ATM and ATR. Progress in biophysics and molecular biology 25 31255703
2018 Kinase-dependent structural role of DNA-PKcs during immunoglobulin class switch recombination. Proceedings of the National Academy of Sciences of the United States of America 25 30072430
2021 CRL4ADTL degrades DNA-PKcs to modulate NHEJ repair and induce genomic instability and subsequent malignant transformation. Oncogene 24 33627782
2020 DNA-PK, Nuclear mTOR, and the Androgen Pathway in Prostate Cancer. Trends in cancer 24 32209447
2020 Activation of DNA-PK by hairpinned DNA ends reveals a stepwise mechanism of kinase activation. Nucleic acids research 24 32716029
2016 DNA-PK Deficiency in Alzheimer's Disease. Journal of neurology & neuromedicine 24 27376156
2022 Dynamics of the Artemis and DNA-PKcs Complex in the Repair of Double-Strand Breaks. Journal of molecular biology 22 36270581
2011 DNA repair gene XRCC7 polymorphisms (rs#7003908 and rs#10109984) and hepatocellular carcinoma related to AFB1 exposure among Guangxi population, China. Hepatology research : the official journal of the Japan Society of Hepatology 22 21883743
2024 DNA-PKcs suppresses illegitimate chromosome rearrangements. Nucleic acids research 21 38412274
2022 DNA-PK Inhibition and Radiation Promote Antitumoral Immunity through RNA Polymerase III in Pancreatic Cancer. Molecular cancer research : MCR 21 35348737
2018 Two siblings with PRKDC defect who presented with cutaneous granulomas and review of the literature. Clinical immunology (Orlando, Fla.) 21 30121298
2019 DNA-PKcs modulates progenitor cell proliferation and fibroblast senescence in idiopathic pulmonary fibrosis. BMC pulmonary medicine 20 31464599
2023 DNA-PKcs and ATM modulate mitochondrial ADP-ATP exchange as an oxidative stress checkpoint mechanism. The EMBO journal 19 36727301
2022 Physical ARTEMIS:DNA-PKcs interaction is necessary for V(D)J recombination. Nucleic acids research 19 35150269
2021 Long noncoding RNA lnc-LEMGC combines with DNA-PKcs to suppress gastric cancer metastasis. Cancer letters 19 34626692
2017 Synergy between Prkdc and Trp53 regulates stem cell proliferation and GI-ARS after irradiation. Cell death and differentiation 19 28686579
2016 micorRNA-101 silences DNA-PKcs and sensitizes pancreatic cancer cells to gemcitabine. Biochemical and biophysical research communications 19 27988337
2004 Expression of DNA-PKcs and Ku86, but not Ku70, differs between lymphoid malignancies. Experimental and molecular pathology 19 15215044

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