Affinage

H2AX

Histone H2AX · UniProt P16104

Length
143 aa
Mass
15.1 kDa
Annotated
2026-04-28
100 papers in source corpus 39 papers cited in narrative 38 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

H2AX is a histone H2A variant that serves as the central chromatin-based platform for the DNA damage response, undergoing rapid phosphorylation at serine 139 (forming γ-H2AX) by ATM, DNA-PK, or ATR kinases across megabase-scale chromatin domains shaped by three-dimensional chromosomal topology, thereby recruiting repair and signaling factors including 53BP1, MDC1, INO80, and BRCA1 (PMID:9488723, PMID:11571274, PMID:12697768, PMID:15607974, PMID:32572033). γ-H2AX is resolved by context-specific phosphatases — PP2A after exogenous DSBs, PP4 after replication stress, and Wip1 as a general attenuator — while additional post-translational modifications including TIP60-mediated acetylation followed by UBC13-dependent ubiquitination, RNF2-BMI1/RNF8-mediated monoubiquitination at K119/K120, SIRT1-regulated K5 deacetylation, EYA-mediated Y142 dephosphorylation, and E141 ADP-ribosylation collectively tune the balance between DNA repair, checkpoint activation, and apoptosis (PMID:16310392, PMID:18614045, PMID:20460517, PMID:17709392, PMID:21676867, PMID:19234442, PMID:33264433, PMID:35258628). Beyond canonical DSB repair, H2AX phosphorylation is required for G2-M checkpoint enforcement, prevention of aberrant CtIP-mediated end resection during V(D)J recombination, meiotic sex chromosome inactivation, oocyte quality surveillance, and DNA ladder formation during apoptosis and necroptosis (PMID:12447390, PMID:21160476, PMID:15589157, PMID:26509888, PMID:16818236, PMID:22972376). H2AX protein levels are post-transcriptionally controlled by miR-24 during hematopoietic differentiation and by a PRMT5–RNF168–SMURF2 proteostasis axis (PMID:19377482, PMID:31533041).

Mechanistic history

Synthesis pass · year-by-year structured walk · 22 steps
  1. 1998 High

    The discovery that DSBs induce rapid phosphorylation of H2AX at S139 across ~2 Mb chromatin domains established γ-H2AX as a large-scale chromatin mark for broken DNA, raising the question of which kinases are responsible and what downstream effectors are recruited.

    Evidence 2D gel electrophoresis and 32P incorporation in irradiated mammalian cells and mice

    PMID:9488723

    Open questions at the time
    • Kinase identity unknown
    • Functional consequence of modification not yet tested
    • Whether phosphorylation is strictly DSB-specific was not addressed
  2. 2001 High

    Identification of ATM as the primary H2AX kinase after DSBs, with DNA-PK providing residual activity, answered the kinase-identity question and established that PIKK family members are the writers of γ-H2AX.

    Evidence In vitro kinase assay, ATM−/− and DNA-PKcs−/− cell lines, rescue by ectopic ATM expression

    PMID:11571274

    Open questions at the time
    • ATR contribution at replication forks not yet tested
    • Relative contributions of ATM vs. DNA-PK in different damage contexts not resolved
  3. 2002 High

    Demonstrating that H2AX−/− mice phenocopy ATM−/− cells for G2-M checkpoint failure at low DSB doses established γ-H2AX as a signal amplifier required for threshold-level checkpoint activation, not merely a damage marker.

    Evidence H2AX−/− and 53BP1−/− mouse knockouts, flow cytometry, 53BP1 foci immunofluorescence

    PMID:12447390

    Open questions at the time
    • Whether H2AX contributes to S-phase or G1 checkpoints was unknown
    • Mechanism of signal amplification not molecularly defined
  4. 2003 High

    Direct binding of 53BP1 to phosphorylated (but not unphosphorylated) H2AX, combined with evidence that DNA-PK phosphorylates H2AX within nucleosomes enhanced by acetylation, established the first molecular link between γ-H2AX and effector recruitment and defined chromatin context as a regulatory layer.

    Evidence In vitro binding assays with H2AX phospho-mutant reconstitution; reconstituted nucleosomal kinase assays

    PMID:12697768 PMID:14627815

    Open questions at the time
    • MDC1 as an intermediary adaptor not yet characterized
    • Whether 53BP1 binding is direct to γ-H2AX or indirect through other marks was debated
  5. 2003 High

    Showing that H2AX haploinsufficiency causes genomic instability and that phosphorylation-site mutants (S→A, S→E) fail to rescue H2AX-null phenotypes established H2AX as a dosage-dependent, phosphorylation-dependent tumor suppressor.

    Evidence H2AX+/−, H2AX−/−, p53−/− double-knockout mice with transgenic phospho-site mutant reconstitution and cytogenetics

    PMID:12914700 PMID:12914701

    Open questions at the time
    • Specific repair pathway(s) compromised by haploinsufficiency not identified
    • Whether phosphomimetic failure reflects structural requirements beyond charge not tested
  6. 2004 High

    ATM and DNA-PK were shown to be redundant for IR-induced γ-H2AX (requiring ablation of both to eliminate signal), while the INO80 chromatin remodeling complex was identified as a γ-H2AX-dependent effector recruited to DSBs, expanding the reader network beyond checkpoint proteins to chromatin remodelers.

    Evidence ATM/DNA-PK double-deficient cells with epistasis analysis; Co-IP and ChIP of INO80 at HO-induced DSBs in yeast

    PMID:15059890 PMID:15607974

    Open questions at the time
    • Whether INO80 recruitment via γ-H2AX is conserved in mammals not shown
    • SWR1/SRCAP remodeler contributions not yet addressed
  7. 2004 High

    Discovery that ATR phosphorylates H2AX on the XY body in a BRCA1-dependent manner for meiotic sex chromosome inactivation extended γ-H2AX function beyond DNA repair to programmed chromatin silencing during gametogenesis.

    Evidence Immunofluorescence of ATR and γ-H2AX in wild-type vs. BRCA1-mutant spermatocytes

    PMID:15589157

    Open questions at the time
    • How ATR-dependent γ-H2AX recruits silencing machinery was not identified
    • Whether H2AX phosphorylation is sufficient for MSCI without other marks not tested
  8. 2005 High

    Identification of PP2A as the phosphatase that dephosphorylates γ-H2AX after repair completion answered how damage foci are resolved and revealed that persistent γ-H2AX impairs repair and cell survival.

    Evidence Co-IP, in vitro phosphatase assay, PP2A RNAi causing persistent γ-H2AX foci and radiosensitivity

    PMID:16310392

    Open questions at the time
    • Whether PP2A acts on nucleosomal γ-H2AX or requires eviction not tested
    • Context-specificity relative to replication stress not addressed
  9. 2006 High

    Demonstrating that DNA-PK (not ATM) phosphorylates H2AX during apoptosis, and that JNK-mediated H2AX phosphorylation is required for DNA ladder formation by CAD, established distinct kinase usage in apoptotic versus repair contexts and a pro-death effector role for γ-H2AX.

    Evidence DNA-PKcs−/− and ATM−/− cells in apoptotic conditions; H2AX−/− MEFs with in vitro CAD assay

    PMID:16567133 PMID:16818236

    Open questions at the time
    • How γ-H2AX facilitates CAD-mediated cleavage structurally was not defined
    • Whether this applies to all apoptotic stimuli was not tested
  10. 2007 High

    Elucidation of a sequential TIP60 acetylation → UBC13 ubiquitination cascade on H2AX that promotes its eviction from damaged chromatin revealed that γ-H2AX removal is an actively regulated process requiring ordered PTMs.

    Evidence In vivo acetylation and ubiquitination assays, TIP60/UBC13 Co-IP, dominant-negative and knockdown experiments

    PMID:17709392

    Open questions at the time
    • Specific acetylation site on H2AX not mapped
    • Whether eviction is required for repair completion or just foci resolution was unclear
  11. 2008 High

    Identification of FACT as the H2AX nucleosome exchange factor stimulated by DNA-PK phosphorylation, PP4 as the replication-stress-specific γ-H2AX phosphatase, and Rvb1-TIP60 as the HAT complex required for γ-H2AX removal collectively established that γ-H2AX dynamics are governed by context-specific writer-eraser-remodeler modules.

    Evidence In vitro nucleosome exchange with purified FACT; PP4 phosphatase assay on mononucleosomes with replication-stress specificity; Rvb1 RNAi epistasis with TIP60

    PMID:18285460 PMID:18406329 PMID:18614045

    Open questions at the time
    • How FACT discriminates H2AX-containing vs. canonical H2A nucleosomes not defined
    • Whether PP4 and PP2A act on distinct pools simultaneously was not tested
  12. 2009 High

    Discovery that EYA dephosphorylates H2AX Y142 to switch the γ-H2AX platform from apoptosis to repair, and that miR-24 post-transcriptionally silences H2AX during differentiation, established two new regulatory layers — a PTM-encoded fate switch and a microRNA-based expression control mechanism.

    Evidence In vitro EYA phosphatase assay, mass spectrometry for Y142, genetic epistasis; miR-24 3′UTR reporter with rescue by miR-24-insensitive H2AX

    PMID:19234442 PMID:19377482

    Open questions at the time
    • WSTF kinase (Y142 writer) interaction with EYA not mechanistically dissected
    • Whether miR-24 regulation is conserved beyond hematopoiesis not shown
  13. 2010 High

    Showing that H2AX prevents CtIP-mediated aberrant end resection at RAG-generated hairpin ends in G1 lymphocytes revealed a cell-cycle- and context-specific protective function of γ-H2AX-MDC1 in enforcing faithful NHEJ during V(D)J recombination.

    Evidence H2AX−/− lymphocytes with CtIP/MDC1 genetic epistasis, V(D)J junction sequencing, cytogenetics

    PMID:21160476

    Open questions at the time
    • Whether this protective mechanism extends to class-switch recombination junctions was not shown
    • How γ-H2AX physically blocks CtIP access was not defined
  14. 2011 High

    Identification of RNF2-BMI1 as the E3 ligase for H2AX K119/K120 monoubiquitination, required for ATM recruitment and full γ-H2AX signaling, established ubiquitination as a prerequisite for (not just consequence of) the DSB response.

    Evidence Co-IP, ubiquitination assay, K119R/K120R mutants, siRNA with DSB marker readouts

    PMID:21676867

    Open questions at the time
    • Whether RNF2-BMI1 acts before or after initial S139 phosphorylation was not resolved temporally
    • Relationship to RNF8/RNF168-mediated ubiquitin signaling not fully distinguished
  15. 2012 High

    Demonstrating that AIF forms a nuclear DNA-degrading complex with γ-H2AX during necroptosis, dependent on ATM/DNA-PK-mediated S139 phosphorylation, established a role for H2AX in caspase-independent programmed cell death.

    Evidence H2AX−/− cells, kinase inhibitors, S139A/S139E mutant reconstitution, AIF-γ-H2AX Co-IP

    PMID:22972376

    Open questions at the time
    • How AIF recognizes γ-H2AX specifically (binding interface) was not defined
    • Whether this mechanism operates in vivo during pathological necroptosis not shown
  16. 2014 High

    Identification of Dub3 as a deubiquitinase that directly removes ubiquitin from H2AX, counteracting RNF8/RNF168, and selectively suppressing 53BP1/BRCA1 foci without affecting MDC1 or γ-H2AX, revealed a deubiquitination-based mechanism for fine-tuning downstream effector recruitment.

    Evidence In vitro DUB assay, catalytically inactive Dub3 mutant, Co-IP, immunofluorescence

    PMID:24704006

    Open questions at the time
    • Whether Dub3 acts on mono- or poly-ubiquitinated H2AX preferentially not determined
    • Physiological context regulating Dub3 activity at DSBs unknown
  17. 2015 High

    Genetic rescue of oocyte loss in XO mice by H2AX deletion or point mutation established γ-H2AX as a functional driver of oocyte elimination triggered by asynapsed chromosomes, extending H2AX function to meiotic quality control in females.

    Evidence H2AX−/− and point-mutant mice on XO background, oocyte counting, γ-H2AX immunofluorescence

    PMID:26509888

    Open questions at the time
    • Downstream effectors linking γ-H2AX to oocyte apoptosis not identified
    • Whether this mechanism applies to autosomal asynapsis was not tested
  18. 2016 Medium

    Chronic oxidative stress was shown to promote RNF168-mediated poly-ubiquitination and proteasomal degradation of H2AX, while γ-H2AX signaling was found to drive chromosome territory relocation via nuclear myosin 1, revealing proteostatic vulnerability and a mechanical effector function of γ-H2AX.

    Evidence ROS modulation with RNF168 Co-IP and ubiquitination assays; H2AX-deficient cells with FISH and NM1 motor-dead mutants

    PMID:27006338 PMID:27365048

    Open questions at the time
    • Whether RNF168-dependent H2AX degradation is a general stress response or tumor-specific is unclear
    • How γ-H2AX recruits NM1 mechanistically is not defined
    • Single-lab findings for chromosome territory relocation
  19. 2018 High

    Conditional MOF deletion in spermatocytes demonstrated that H4K16 acetylation by MOF is required for chromatin-wide γ-H2AX spreading during all three meiotic waves, distinguishing axis-restricted from chromatin-wide γ-H2AX as functionally distinct states.

    Evidence Germ cell-specific Mof knockout, chromosome spreading, γ-H2AX and MDC1 immunofluorescence

    PMID:29795555

    Open questions at the time
    • Whether MOF acts by enabling ATR/ATM access or by facilitating MDC1 spreading not distinguished
    • Applicability to somatic DSB-induced γ-H2AX spreading not tested
  20. 2019 Medium

    The PRMT5–RNF168–SMURF2 axis was identified as a proteostasis circuit controlling H2AX protein levels, where PRMT5 sustains RNF168 to stabilize H2AX while SMURF2 promotes its degradation, linking epigenetic enzyme activity to H2AX abundance.

    Evidence Co-IP, RNAi, protein stability assays in MTAP-deficient glioblastoma cells

    PMID:31533041

    Open questions at the time
    • Whether SMURF2-mediated degradation is ubiquitin-dependent at the same sites as RNF168 mono-ub was not clarified
    • Single-lab study in a specific genetic context (MTAP-deleted)
    • In vivo validation lacking
  21. 2020 High

    Hi-C-resolved mapping of γ-H2AX domains showed that spreading follows pre-existing 3D chromosomal contacts rather than linear distance, while ADP-ribosylation at E141 was discovered to direct BER and suppress erroneous DSB signaling, together refining the spatial and PTM logic of H2AX-based damage signaling.

    Evidence Hi-C + ChIP-seq at defined DSBs; mass spectrometry identification of E141 ADP-ribosylation with site mutant and Neil3 Co-IP

    PMID:32572033 PMID:33264433

    Open questions at the time
    • Whether cohesin or condensin mediate the contact-dependent spreading was not tested
    • Which ADP-ribosyltransferase modifies E141 was not identified
  22. 2023 High

    SIRT1 was identified as the deacetylase for H2AX K5, and K5 acetylation was shown to impair S139 phosphorylation, establishing a crosstalk in which acetylation state gates the canonical γ-H2AX response.

    Evidence Cardiomyocyte-specific Sirt1 knockout, H2AX K5Q and S139A mutants, acetyl-K5 and phospho-S139 immunostaining

    PMID:35258628

    Open questions at the time
    • The acetyltransferase that writes K5 acetylation was not identified
    • Whether K5 acetylation inhibits kinase access or promotes phosphatase activity not distinguished

Open questions

Synthesis pass · forward-looking unresolved questions
  • Key unresolved questions include: the structural basis for how γ-H2AX recruits each specific effector (53BP1, MDC1, AIF); how the multiple PTMs on H2AX (S139-P, Y142-P, K5-Ac, K119/120-Ub, E141-ADP-ribose) are coordinated temporally and spatially in a single nucleosome; and whether the non-canonical roles of H2AX in stem cell proliferation and DNA demethylation represent generalizable mechanisms or context-restricted phenomena.
  • No cryo-EM or crystal structure of γ-H2AX nucleosome with bound effector
  • Combinatorial PTM code on single H2AX molecules not resolved
  • In vivo relevance of GABA receptor–H2AX axis and HMGA2–FACT–H2AX demethylation pathway requires independent replication

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0042393 histone binding 4 GO:0005198 structural molecule activity 3
Localization
GO:0005694 chromosome 5 GO:0000228 nuclear chromosome 3 GO:0005634 nucleus 3
Pathway
R-HSA-73894 DNA Repair 11 R-HSA-4839726 Chromatin organization 7 R-HSA-1474165 Reproduction 3 R-HSA-1640170 Cell Cycle 3 R-HSA-5357801 Programmed Cell Death 3 R-HSA-168256 Immune System 1
Partners
ATMATRDNA-PKCSMDC1RNF168SMURF2TIP60TP53BP1
Complex memberships
nucleosome

Evidence

Reading pass · 38 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1998 DNA double-strand breaks induce rapid phosphorylation of histone H2AX specifically at serine 139 (forming γ-H2AX), with approximately 2×10⁶ base pairs of chromatin involved per DSB, indicating large-scale chromatin domain modification at each break site. Two-dimensional gel electrophoresis, 32P incorporation, in vivo phosphorylation assays in irradiated mammalian cells and mice The Journal of biological chemistry High 9488723
2001 ATM is the major kinase responsible for H2AX phosphorylation at serine 139 in response to DSBs; ATM phosphorylates H2AX in vitro, H2AX phosphorylation is severely compromised in ATM-/- cells, and ectopic ATM expression in ATM-/- cells restores H2AX phosphorylation. DNA-PK accounts for residual phosphorylation in the absence of ATM. In vitro kinase assay, ATM-/- and DNA-PKcs-/- cell lines, rescue by ectopic ATM expression, wortmannin inhibition The Journal of biological chemistry High 11571274
2004 ATM and DNA-PK function redundantly to phosphorylate H2AX after ionizing radiation; ablation of both kinases is required to eliminate IR-induced H2AX phosphorylation. ATR is not involved in H2AX phosphorylation in non-replicating cells. DNA-PK-mediated H2AX phosphorylation contributes to recruitment of MDC1 and 53BP1 to DSB sites. ATM-deficient cells treated with LY294002 (DNA-PK inhibitor); ATM/DNA-PK double-deficient chicken cells; epistasis analysis Cancer research High 15059890
2002 H2AX is required for efficient G2-M checkpoint activation after low doses of ionizing radiation; H2AX-/- mice show a G2-M checkpoint defect comparable to ATM-/- cells. H2AX regulates the efficient accumulation of 53BP1 into IR-induced foci, thereby amplifying the damage signal at threshold DSB levels. H2AX-/- and 53BP1-/- mouse genetic knockouts, flow cytometry cell cycle analysis, immunofluorescence for 53BP1 foci Nature cell biology High 12447390
2003 53BP1 directly binds phosphorylated H2AX (but not unphosphorylated H2AX) through a region upstream of its C-terminus; H2AX phosphorylation at serine 139/140 is required for efficient 53BP1 foci formation at DNA breaks. Hyperphosphorylation and relocalization of 53BP1 occur independently. In vitro binding assays, H2AX-deficient cells reconstituted with wild-type or phosphorylation-deficient H2AX mutants, immunofluorescence The Journal of biological chemistry High 12697768
2003 H2AX functions as a dosage-dependent suppressor of genomic instability and tumors; H2AX haploinsufficiency causes genomic instability in normal cells. Restoration of H2AX null allele with wild-type H2AX restores genomic stability and radiation resistance, but substitution of the conserved serine phosphorylation sites (S→A or S→E) abolishes this rescue, demonstrating phosphorylation-dependence. Gene targeting (H2AX-/- and H2AX+/- mice), p53-/- double knockouts, transgenic reconstitution with phosphorylation-site mutants, cytogenetic analysis Cell High 12914700 12914701
2004 The INO80 chromatin remodeling complex is recruited to DSBs through a direct interaction with γ-H2AX (phosphorylated H2AX); this interaction requires the Nhp10 HMG-like subunit of INO80. Loss of Nhp10 or γ-H2AX reduces INO80 recruitment to DSBs, linking ATP-dependent chromatin remodeling to DNA repair. Co-immunoprecipitation, chromatin immunoprecipitation at HO endonuclease-induced DSB in yeast, genetic epistasis with RAD52 pathway Cell High 15607974
2004 BRCA1 is required for recruiting ATR kinase to XY chromatin at the onset of meiotic sex chromosome inactivation (MSCI); ATR then phosphorylates H2AX within the XY body, triggering chromatin condensation and transcriptional repression of sex chromosomes. Immunofluorescence localization of ATR in wild-type vs. BRCA1-mutant pachytene spermatocytes, correlation of ATR localization with H2AX phosphorylation and MSCI Current biology : CB High 15589157
2005 Protein phosphatase 2A (PP2A) dephosphorylates γ-H2AX to resolve DNA damage foci after repair. PP2A catalytic subunit co-immunoprecipitates and co-localizes with γ-H2AX in foci; PP2A dephosphorylates γ-H2AX in vitro; inhibition or RNAi silencing of PP2A causes persistent γ-H2AX foci, inefficient DNA repair, and hypersensitivity to DNA damage. Co-immunoprecipitation, colocalization, in vitro phosphatase assay, RNAi knockdown, clonogenic survival Molecular cell High 16310392
2006 H2AX is phosphorylated by JNK activated by UVA, and this phosphorylation is required for DNA ladder formation (internucleosomal DNA fragmentation) during apoptosis but not for caspase-3 activation. H2AX phosphorylation is critical for DNA degradation by caspase-activated DNase (CAD) in vitro. H2AX-/- mouse embryonic fibroblasts, in vitro CAD assay, JNK kinase assay, caspase activity measurement Molecular cell High 16818236
2006 DNA-PK (not ATM) is solely responsible for H2AX phosphorylation during apoptotic DNA fragmentation; ATM is degraded before DNA fragmentation occurs, while DNA-PKcs is robustly activated (autophosphorylation at S2056) in apoptotic cells before being inactivated by proteolysis. DNA-PKcs-/- and ATM-/- cells, immunoblotting, kinase activity assays in apoptotic vs. non-apoptotic cells DNA repair High 16567133
2007 TIP60 histone acetyltransferase acetylates H2AX following ionizing radiation, and this acetylation is prerequisite for subsequent ubiquitination of H2AX by the TIP60-UBC13 complex. The sequential acetylation and ubiquitination leads to release of H2AX from damaged chromatin, promoting chromatin dynamics required for DNA damage response. In vivo acetylation and ubiquitination assays, TIP60/UBC13 co-immunoprecipitation, dominant negative and knockdown experiments Molecular and cellular biology High 17709392
2008 FACT (Spt16/SSRP1) complex is the major regulator of nucleosomal H2AX exchange; DNA-PK phosphorylation of H2AX facilitates FACT-mediated H2AX exchange by inducing nucleosome conformational changes, while PARP1-mediated poly-ADP-ribosylation of Spt16 inhibits FACT activity for H2AX exchange. Biochemical purification of H2AX-associated factors, in vitro nucleosome exchange assays, Co-IP, inhibitor treatments Molecular cell High 18406329
2008 A PP4 phosphatase complex (PP4C/PP4R2/PP4R3β) specifically dephosphorylates ATR-mediated γ-H2AX generated during DNA replication. PP4 directly dephosphorylates γ-H2AX within mononucleosomes in vitro; PP4 silencing causes persistent replication-associated γ-H2AX foci and hypersensitivity to replication inhibitors but not radiomimetic drugs. In vitro phosphatase assay on mononucleosomes, RNAi knockdown, clonogenic survival with replication inhibitors vs. radiomimetics Molecular cell High 18614045
2009 EYA protein tyrosine phosphatase dephosphorylates H2AX at tyrosine 142 (Y142) of the C-terminal tail. Phosphorylation of Y142 determines whether DNA repair factors or pro-apoptotic factors are recruited to γ-H2AX (phospho-S139); dephosphorylation of Y142 by EYA promotes DNA repair and survival over apoptosis. In vitro phosphatase assay, co-immunoprecipitation, mass spectrometry identification of Y142, genetic epistasis in mammalian cells Nature High 19234442
2009 miR-24 is upregulated during terminal hematopoietic differentiation and suppresses H2AX expression by targeting conserved binding sites in the H2AX 3'UTR. miR-24-mediated H2AX reduction renders terminally differentiated blood cells hypersensitive to DNA damage; this phenotype is rescued by miR-24-insensitive H2AX. miRNA target validation (3'UTR reporter assays), Western blot, γ-irradiation sensitivity assays in differentiated vs. undifferentiated cells, rescue with miR-24-resistant H2AX Nature structural & molecular biology High 19377482
2010 Wip1 phosphatase directly dephosphorylates γ-H2AX in vitro and in vivo; ectopic Wip1 expression reduces γ-H2AX after ionizing radiation, disrupts recruitment of DNA repair factors to damage sites, and delays repair. Wip1 deletion enhances γ-H2AX in cells under oncogenic stress. In vitro phosphatase assay, ectopic Wip1 expression, Wip1 knockout, immunofluorescence for repair factor recruitment Cancer research High 20460517
2010 H2AX prevents CtIP-mediated DNA end resection of hairpin-sealed coding ends in G1-phase lymphocytes. In the absence of H2AX, CtIP can open RAG-generated hairpin ends and resect DNA, leading to aberrant NHEJ using micro-homologies and chromosomal deletions. This protective function of H2AX requires γ-H2AX and MDC1. H2AX-/- mouse lymphocytes, genetic epistasis with CtIP and MDC1, sequencing of V(D)J junctions, cytogenetic analysis Nature High 21160476
2011 Monoubiquitination of H2AX at Lys119/Lys120 mediated by the RNF2-BMI1 E3 ligase complex is required for efficient γ-H2AX formation and DNA damage response signaling. RNF2-BMI1 interacts with H2AX in a DNA damage-dependent manner; H2AX K120R mutation abolishes monoubiquitination, impairs ATM recruitment to DSBs, and reduces MDC1 accumulation. Co-immunoprecipitation, ubiquitination assay, site-directed mutagenesis (K119R/K120R), siRNA knockdown, immunofluorescence for DSB markers The Journal of biological chemistry High 21676867
2003 DNA-PK is activated by nucleosomes through Ku binding to nucleosomal DNA ends, and the activated complex phosphorylates H2AX within the nucleosome context. Histone acetylation greatly enhances DNA-PK-mediated H2AX phosphorylation within nucleosomes but not when H2AX is in free form. In vitro kinase assays with reconstituted nucleosomes, Ku binding assays, comparison of acetylated vs. non-acetylated nucleosomal substrates Nucleic acids research High 14627815
2005 ATM-dependent phosphorylation of H2AX occurs during mitosis in normally growing (unirradiated) mammalian cells, revealing a DNA damage-independent role for γ-H2AX. Two distinct focal populations of γ-H2AX exist: large amorphous foci recruiting DSB repair proteins and smaller foci that do not recruit repair proteins. Quantitative in situ immunofluorescence, cell cycle analysis, ATM inhibition in unirradiated cells Molecular biology of the cell Medium 16030261
2008 GABA(A) receptor signaling in embryonic stem cells signals through S-phase checkpoint kinases (PIKKs) and histone H2AX to regulate stem cell proliferation, independent of differentiation, apoptosis, or DNA damage; H2AX functions as an effector downstream of GABA(A) receptor-induced hyperpolarization to control S-phase checkpoint-mediated proliferation. GABA(A) receptor pharmacology, flow cytometry, genetic manipulation of H2AX in ES and neural crest stem cells Nature Medium 18515516
2009 H2AX overexpression activates Nox1-mediated ROS generation through a pathway involving Rac1 GTPase; H2AX reduces the interaction between Nox1 activator NOXA1 and its inhibitor 14-3-3ζ, thereby increasing Nox1 activity and promoting cell death after DNA damage. H2AX overexpression and knockdown, Nox1 activity assays, Rac1 dominant-negative expression, NAC treatment, Co-IP of NOXA1/14-3-3ζ Cell death & disease Medium 22237206
2008 Rvb1 is required for the histone acetyltransferase activity of the Tip60/NuA4 complex; Rvb1 depletion mimics Tip60 depletion in causing persistence of phospho-H2AX after DNA damage. H4 acetylation by Tip60 is required prior to γ-H2AX dephosphorylation, linking the Rvb1-Tip60 complex to γ-H2AX removal. RNAi knockdown of Rvb1 vs. Ino80 vs. SRCAP vs. Tip60, HAT activity assay, γ-H2AX immunofluorescence Molecular and cellular biology Medium 18285460
2009 H2AX is required for cell cycle arrest via the p53/p21 pathway after replication stalling; absence of H2AX leads to proteasome-dependent p21 degradation followed by caspase-dependent mitotic catastrophe, while H2AX-proficient cells increase p21 and arrest the cell cycle. H2AX-/- cells, RNAi, adeno-associated virus model of pannuclear γ-H2AX, proteasome inhibitors, p21 immunoblotting Molecular and cellular biology Medium 19273588
2008 ATR and H2AX cooperate to maintain genome stability under replication stress; in ATR-deficient cells, H2AX is phosphorylated by ATM and DNA-PKcs and promotes Rad51 focal accumulation (homologous recombination); dual ATR/H2AX deficiency causes synergistic increases in chromatid breaks and translocations. The S139 phosphorylation site of H2AX is specifically required. ATR-deficient cells combined with H2AX-/- cells, Rad51 immunofluorescence, cytogenetic analysis, H2AX S139A mutant reconstitution The Journal of biological chemistry High 19049966
2012 AIF-mediated caspase-independent necroptosis requires γ-H2AX (S139-phosphorylated H2AX); AIF associates with γ-H2AX in the nucleus to form a DNA-degrading complex. ATM and DNA-PK synergistically phosphorylate H2AX at S139 to enable necroptosis; H2AX S139A mutant or H2AX-/- cells are resistant to necroptosis, while phosphomimetic H2AX-S139E restores sensitivity. H2AX-/- cells, ATM/DNA-PK inhibitors, H2AX S139A and S139E mutant reconstitution, Co-IP of AIF/γ-H2AX Cell death & disease High 22972376
2014 Dub3 deubiquitinase directly deubiquitinates H2AX; Dub3 overexpression decreases DNA damage-induced H2AX monoubiquitination and abrogates 53BP1 and BRCA1 focus formation (but not MDC1 or γ-H2AX foci), while Dub3 depletion has the opposite effect. Dub3 counteracts the H2AX E3 ligases RNF8 and RNF168. In vitro deubiquitinase assay, Co-IP of Dub3/H2AX, overexpression of wild-type vs. catalytically inactive Dub3, RNAi, immunofluorescence Molecular oncology High 24704006
2019 PRMT5 sustains RNF168 expression; suppression of PRMT5 (in MTAP-deficient cells) reduces RNF168 levels, leading to H2AX destabilization by E3 ubiquitin ligase SMURF2. RNF168 and SMURF2 serve as a stabilizer and destabilizer of H2AX respectively through dynamic interactions with H2AX, forming a proteostasis regulatory axis. Co-immunoprecipitation, RNAi knockdown, protein stability assays, MTAP-deficient glioblastoma cells Cell reports Medium 31533041
2020 H2AX glutamate 141 (E141) is ADP-ribosylated following oxidative DNA damage; this modification recruits Neil3 glycosylase to DNA damage sites for base excision repair. Loss of E141 ADP-ribosylation enhances S139 phosphorylation (γH2AX) and erroneously triggers DSB response factors, indicating ADP-ribosylation suppresses the DSB response during BER. Unbiased mass spectrometry identification of ADP-ribosylation site, H2AX E141A mutant, Neil3 Co-IP, BER functional assays The EMBO journal High 33264433
2016 Chronic oxidative stress promotes H2AX poly-ubiquitination by RNF168 and subsequent proteasomal degradation; persistent ROS (due to deficient JunD/Nrf2 antioxidant response) enhances H2AX-RNF168 interaction and H2AX turnover, reducing DNA repair capacity. Co-immunoprecipitation of H2AX-RNF168, ubiquitination assays, proteasome inhibition, ROS modulation, primary TNBC patient samples EMBO molecular medicine Medium 27006338
2023 SIRT1 deacetylates H2AX at Lys5; acetylation of H2AX at K5 (mimicked by K5Q mutation) impairs Ser139 phosphorylation in response to DNA damage. SIRT1 deficiency in cardiomyocytes elevates K5 acetylation of H2AX and blunts Ser139 phosphorylation, enhancing doxorubicin-induced cardiotoxicity. Cardiomyocyte-specific Sirt1 knockout mice, H2AX K5Q and S139A mutants, immunostaining for acetyl-K5-H2AX and phospho-S139-H2AX, caspase-3 activation assay Cardiovascular research High 35258628
2020 γH2AX domains spread primarily along chromosomal contacts of a DSB site as determined by topological architecture; DSBs that disrupt a topological border allow γH2AX to extend into both adjacent compartments, while DSBs near a border produce highly asymmetric γH2AX platforms with near-absence from one broken end. Hi-C/chromatin conformation capture combined with ChIP-seq for γH2AX at defined DSBs Nature communications High 32572033
2021 HMGA2 induces DNA nicks at transcription start sites, which enables the FACT complex to incorporate H2AX-containing nucleosomes; phosphorylation of H2AX at S139 (γH2AX) at these sites is required for repair-mediated active DNA demethylation and transcription activation. ChIP-seq, HMGA2 knockdown, FACT complex inhibition, H2AX phosphorylation site mutants, bisulfite sequencing for DNA methylation Nature communications Medium 33594057
2020 UBE2T-RNF8 E2-E3 ubiquitin ligase pair monoubiquitinates H2AX/γH2AX at K119/K120 upon radiation; this monoubiquitination facilitates CHK1 phosphorylation and CHK1 release from chromatin for activation. H2AX K119/120R mutation abolishes monoubiquitination and abrogates CHK1 activation. Co-immunoprecipitation, ubiquitination assays, chromatin fractionation, H2AX K119/120R mutant, CHK1 activation assays, xenograft models Journal of experimental & clinical cancer research Medium 33087136
2015 In female mice with chromosome abnormalities (e.g., XO Turner syndrome), asynapsed chromosomes during meiotic prophase I trigger oocyte elimination at diplonema linked to γH2AX presence; deletion or point mutation of H2afx restores oocyte numbers in XO females to wild-type (XX) levels, establishing H2AX phosphorylation as a driver (not merely marker) of oocyte elimination. H2afx-/- and H2afx point-mutant mice on XO background, immunofluorescence for γH2AX, oocyte counting at diplonema PLoS genetics High 26509888
2018 Histone acetyltransferase MOF regulates the expansion of H2AX phosphorylation (γH2AX) in spermatocytes during all three meiotic waves (leptonema, zygonema, pachynema). Germ cell-specific Mof deletion causes loss of H4K16 acetylation, restricts γH2AX to chromosomal axes without chromatin-wide expansion, and causes MSCI failure and DSB repair defects. Germ cell-specific Mof knockout (Stra8-Cre), immunofluorescence for γH2AX and MDC1, chromosome spreading, crossover analysis PLoS genetics High 29795555
2016 γH2AX signaling is a prerequisite for damage-induced chromosome territory relocation; cells deficient in γH2AX signaling fail to relocate chromosome territories after DSBs. γH2AX signaling promotes nuclear myosin 1 (NM1) recruitment to chromatin, and NM1 motor function is essential for chromosome territory movement. H2AX-deficient cells, immunofluorescence for chromosome territories (FISH), NM1 motor-dead mutants, chromatin fractionation Nucleic acids research Medium 27365048

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1998 DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139. The Journal of biological chemistry 4387 9488723
2001 ATM phosphorylates histone H2AX in response to DNA double-strand breaks. The Journal of biological chemistry 1574 11571274
2008 Gamma-H2AX - a novel biomarker for DNA double-strand breaks. In vivo (Athens, Greece) 1062 18610740
2008 Gamma-H2AX in recognition and signaling of DNA double-strand breaks in the context of chromatin. Nucleic acids research 1013 18772227
2004 ATM and DNA-PK function redundantly to phosphorylate H2AX after exposure to ionizing radiation. Cancer research 826 15059890
2002 Histone H2A variants H2AX and H2AZ. Current opinion in genetics & development 591 11893489
2012 Histone H2AX phosphorylation: a marker for DNA damage. Methods in molecular biology (Clifton, N.J.) 567 22941631
2002 DNA damage-induced G2-M checkpoint activation by histone H2AX and 53BP1. Nature cell biology 565 12447390
2010 H2AX Phosphorylation: Its Role in DNA Damage Response and Cancer Therapy. Journal of nucleic acids 474 20811597
2003 H2AX haploinsufficiency modifies genomic stability and tumor susceptibility. Cell 464 12914701
2005 gamma-H2AX dephosphorylation by protein phosphatase 2A facilitates DNA double-strand break repair. Molecular cell 449 16310392
2004 INO80 and gamma-H2AX interaction links ATP-dependent chromatin remodeling to DNA damage repair. Cell 448 15607974
2009 Tyrosine dephosphorylation of H2AX modulates apoptosis and survival decisions. Nature 438 19234442
2003 Histone H2AX: a dosage-dependent suppressor of oncogenic translocations and tumors. Cell 404 12914700
2003 Characteristics of gamma-H2AX foci at DNA double-strand breaks sites. Biochemistry and cell biology = Biochimie et biologie cellulaire 367 12897845
2004 BRCA1, histone H2AX phosphorylation, and male meiotic sex chromosome inactivation. Current biology : CB 325 15589157
2006 Cell apoptosis: requirement of H2AX in DNA ladder formation, but not for the activation of caspase-3. Molecular cell 311 16818236
2003 Accumulation of checkpoint protein 53BP1 at DNA breaks involves its binding to phosphorylated histone H2AX. The Journal of biological chemistry 301 12697768
2007 DNA damage-dependent acetylation and ubiquitination of H2AX enhances chromatin dynamics. Molecular and cellular biology 295 17709392
2015 Multiple facets of histone variant H2AX: a DNA double-strand-break marker with several biological functions. Nucleic acids research 285 25712102
2003 Expression of phosphorylated histone H2AX in cultured cell lines following exposure to X-rays. International journal of radiation biology 277 12943243
2009 H2AX: functional roles and potential applications. Chromosoma 263 19707781
2012 DNA damage induces reactive oxygen species generation through the H2AX-Nox1/Rac1 pathway. Cell death & disease 248 22237206
2004 Phosphorylation of histone H2AX as a measure of radiosensitivity. International journal of radiation oncology, biology, physics 246 14751500
2010 Focus on histone variant H2AX: to be or not to be. FEBS letters 242 20493860
2003 Histone H2AX phosphorylation as a predictor of radiosensitivity and target for radiotherapy. The Journal of biological chemistry 241 14561744
2009 miR-24-mediated downregulation of H2AX suppresses DNA repair in terminally differentiated blood cells. Nature structural & molecular biology 233 19377482
2008 Histone H2AX-dependent GABA(A) receptor regulation of stem cell proliferation. Nature 233 18185516
2005 ATM-dependent DNA damage-independent mitotic phosphorylation of H2AX in normally growing mammalian cells. Molecular biology of the cell 222 16030261
2008 FACT-mediated exchange of histone variant H2AX regulated by phosphorylation of H2AX and ADP-ribosylation of Spt16. Molecular cell 203 18406329
2008 A PP4-phosphatase complex dephosphorylates gamma-H2AX generated during DNA replication. Molecular cell 200 18614045
2013 Double strand break repair functions of histone H2AX. Mutation research 196 23916969
2004 H2AX may function as an anchor to hold broken chromosomal DNA ends in close proximity. Cell cycle (Georgetown, Tex.) 196 14712078
2005 Chromatin in need of a fix: phosphorylation of H2AX connects chromatin to DNA repair. Molecular cell 178 15949437
2007 Cytometry of ATM activation and histone H2AX phosphorylation to estimate extent of DNA damage induced by exogenous agents. Cytometry. Part A : the journal of the International Society for Analytical Cytology 177 17622968
2006 DNA-PK phosphorylates histone H2AX during apoptotic DNA fragmentation in mammalian cells. DNA repair 175 16567133
2003 DNA double-strand breaks and gamma-H2AX signaling in the testis. Biology of reproduction 174 12533428
2006 Constitutive histone H2AX phosphorylation and ATM activation, the reporters of DNA damage by endogenous oxidants. Cell cycle (Georgetown, Tex.) 173 16940754
2007 H2AX phosphorylation marks gemcitabine-induced stalled replication forks and their collapse upon S-phase checkpoint abrogation. Molecular cancer therapeutics 167 17406032
2006 Induction and quantification of gamma-H2AX foci following low and high LET-irradiation. International journal of radiation biology 166 16546909
2009 gamma-H2AX as protein biomarker for radiation exposure. Annali dell'Istituto superiore di sanita 165 19861731
2020 DNA double-strand breaks induce H2Ax phosphorylation domains in a contact-dependent manner. Nature communications 164 32572033
2005 DNA repair: the importance of phosphorylating histone H2AX. Current biology : CB 161 15694301
2009 H2AX is required for cell cycle arrest via the p53/p21 pathway. Molecular and cellular biology 142 19273588
2011 Monoubiquitination of H2AX protein regulates DNA damage response signaling. The Journal of biological chemistry 141 21676867
2016 Chronic oxidative stress promotes H2AX protein degradation and enhances chemosensitivity in breast cancer patients. EMBO molecular medicine 137 27006338
2008 Human Rvb1/Tip49 is required for the histone acetyltransferase activity of Tip60/NuA4 and for the downregulation of phosphorylation on H2AX after DNA damage. Molecular and cellular biology 130 18285460
2010 Wip1 directly dephosphorylates gamma-H2AX and attenuates the DNA damage response. Cancer research 128 20460517
2003 DNA-PK is activated by nucleosomes and phosphorylates H2AX within the nucleosomes in an acetylation-dependent manner. Nucleic acids research 125 14627815
2009 Kinetics of H2AX phosphorylation after exposure to cisplatin. Cytometry. Part B, Clinical cytometry 124 18727058
2011 MicroRNA-138 modulates DNA damage response by repressing histone H2AX expression. Molecular cancer research : MCR 121 21693595
2010 H2AX prevents CtIP-mediated DNA end resection and aberrant repair in G1-phase lymphocytes. Nature 119 21160476
2011 H2AX phosphorylation at the sites of DNA double-strand breaks in cultivated mammalian cells and tissues. Clinical epigenetics 111 22704343
2008 ATR and H2AX cooperate in maintaining genome stability under replication stress. The Journal of biological chemistry 109 19049966
2024 H2AX: A key player in DNA damage response and a promising target for cancer therapy. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 106 38688170
2008 Complementary functions of ATM and H2AX in development and suppression of genomic instability. Proceedings of the National Academy of Sciences of the United States of America 98 18599436
2014 The nuclear γ-H2AX apoptotic ring: implications for cancers and autoimmune diseases. Cellular and molecular life sciences : CMLS 93 24448903
2006 H2AX chromatin structures and their response to DNA damage revealed by 4Pi microscopy. Proceedings of the National Academy of Sciences of the United States of America 91 17110439
2020 UBE2T-regulated H2AX monoubiquitination induces hepatocellular carcinoma radioresistance by facilitating CHK1 activation. Journal of experimental & clinical cancer research : CR 84 33087136
2009 MRE11-RAD50-NBS1 complex dictates DNA repair independent of H2AX. The Journal of biological chemistry 84 19910469
2012 γ-H2AX and other histone post-translational modifications in the clinic. Biochimica et biophysica acta 83 22430255
2012 BCLAF1 is a radiation-induced H2AX-interacting partner involved in γH2AX-mediated regulation of apoptosis and DNA repair. Cell death & disease 81 22833098
2012 AIF-mediated caspase-independent necroptosis requires ATM and DNA-PK-induced histone H2AX Ser139 phosphorylation. Cell death & disease 76 22972376
2006 Techniques for gamma-H2AX detection. Methods in enzymology 75 16793405
2008 Role of H2AX in DNA damage response and human cancers. Mutation research 74 18804552
2017 Phosphorylated fraction of H2AX as a measurement for DNA damage in cancer cells and potential applications of a novel assay. PloS one 72 28158293
2014 γ-H2AX as a biomarker for DNA double-strand breaks in ecotoxicology. Ecotoxicology and environmental safety 65 24780228
2023 SIRT1 in the cardiomyocyte counteracts doxorubicin-induced cardiotoxicity via regulating histone H2AX. Cardiovascular research 59 35258628
2020 Potential application of γ-H2AX as a biodosimetry tool for radiation triage. Mutation research. Reviews in mutation research 58 34083048
2015 Histone H2AFX Links Meiotic Chromosome Asynapsis to Prophase I Oocyte Loss in Mammals. PLoS genetics 58 26509888
2015 Complementarity of phosphorylated histones H2AX and H3 quantification in different cell lines for genotoxicity screening. Archives of toxicology 53 26404763
2007 Phosphorylation of histone H2AX in radiation-induced micronuclei. Radiation research 50 17903033
2010 H2AX post-translational modifications in the ionizing radiation response and homologous recombination. Cell cycle (Georgetown, Tex.) 49 20703100
2009 Mechanism of elimination of phosphorylated histone H2AX from chromatin after repair of DNA double-strand breaks. Mutation research 49 19682466
2010 Structure and function of histone H2AX. Sub-cellular biochemistry 48 20012577
2021 Positioning of nucleosomes containing γ-H2AX precedes active DNA demethylation and transcription initiation. Nature communications 44 33594057
2019 MDC1 PST-repeat region promotes histone H2AX-independent chromatin association and DNA damage tolerance. Nature communications 44 31729360
2014 Gamma histone 2AX (γ-H2AX)as a predictive tool in radiation oncology. Biomarkers : biochemical indicators of exposure, response, and susceptibility to chemicals 44 24611829
2020 ADP-ribosylation of histone variant H2AX promotes base excision repair. The EMBO journal 43 33264433
2016 Chromosome territory relocation during DNA repair requires nuclear myosin 1 recruitment to chromatin mediated by ϒ-H2AX signaling. Nucleic acids research 43 27365048
2010 Conserved gammaherpesvirus kinase and histone variant H2AX facilitate gammaherpesvirus latency in vivo. Virology 42 20557919
2018 Monoubiquitinated γ-H2AX: Abundant product and specific biomarker for non-apoptotic DNA double-strand breaks. Toxicology and applied pharmacology 41 30006243
2005 DNA-PK is responsible for enhanced phosphorylation of histone H2AX under hypertonic conditions. DNA repair 41 16046194
2015 Biodosimetry Based on γ-H2AX Quantification and Cytogenetics after Partial- and Total-Body Irradiation during Fractionated Radiotherapy. Radiation research 37 25844946
2019 A PRMT5-RNF168-SMURF2 Axis Controls H2AX Proteostasis. Cell reports 36 31533041
2007 Genetic variation in H2AFX contributes to risk of non-Hodgkin lymphoma. Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology 36 17548670
2003 Expression and radiation-induced phosphorylation of histone H2AX in mammalian cells. Journal of radiation research 36 12841599
2021 The Phosphorylated Form of the Histone H2AX (γH2AX) in the Brain from Embryonic Life to Old Age. Molecules (Basel, Switzerland) 35 34885784
2008 Soluble histone H2AX is induced by DNA replication stress and sensitizes cells to undergo apoptosis. Molecular cancer 35 18616816
2014 Dub3 controls DNA damage signalling by direct deubiquitination of H2AX. Molecular oncology 34 24704006
2021 Detection and quantification of γ-H2AX using a dissociation enhanced lanthanide fluorescence immunoassay. Scientific reports 33 33903655
2014 Phosphorylation of histone H2AX in the mouse brain from development to senescence. International journal of molecular sciences 33 24451138
2013 HIF mediated and DNA damage independent histone H2AX phosphorylation in chronic hypoxia. Biological chemistry 33 23241668
2008 The HINT1 tumor suppressor regulates both gamma-H2AX and ATM in response to DNA damage. The Journal of cell biology 33 18852295
2018 MOF influences meiotic expansion of H2AX phosphorylation and spermatogenesis in mice. PLoS genetics 32 29795555
2010 Proteomic dissection of cell type-specific H2AX-interacting protein complex associated with hepatocellular carcinoma. Journal of proteome research 32 20000738
2018 Histone H2AX deficiency causes neurobehavioral deficits and impaired redox homeostasis. Nature communications 30 29670103
2014 Epstein-Barr virus essential antigen EBNA3C attenuates H2AX expression. Journal of virology 30 24429368
2011 Phosphorylation of histone H2AX is a powerful tool for detecting chemical photogenotoxicity. The Journal of investigative dermatology 30 21368771
2008 Copy number alterations of the H2AFX gene in sporadic breast cancer patients. Cancer genetics and cytogenetics 30 18206537