Affinage

ANP32E

Acidic leucine-rich nuclear phosphoprotein 32 family member E · UniProt Q9BTT0

Length
268 aa
Mass
30.7 kDa
Annotated
2026-06-09
30 papers in source corpus 15 papers cited in narrative 14 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 7/7 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

ANP32E is a histone chaperone that specifically removes the histone variant H2A.Z from chromatin to restrict nucleosome dynamics and chromatin accessibility genome-wide (PMID:24463511, PMID:33033242). It recognizes H2A.Z/H2B dimers through a dedicated H2A.Z-interacting domain (ZID) whose residues contact the extended H2A.Z C-terminal αC-helix, a contact resolved at high resolution by crystallography and conferring preference for H2A.Z over canonical H2A (PMID:24463511, PMID:24613878). At active promoters, enhancers, and insulators, ANP32E counteracts H2A.Z deposition—directly antagonizing the SRCAP subunit VPS72 by stabilizing nucleosomes and preventing DNA unwrapping—so that its loss elevates H2A.Z, improves nucleosome positioning, increases transcription factor binding, and raises neighboring gene expression (PMID:33033242, PMID:41646306). The same activity operates in defined biological contexts: ANP32E is rapidly recruited to DNA double-strand breaks where H2A.Z removal permits H4 acetylation and limits CtIP-dependent end resection and alternative non-homologous end joining (PMID:26034280); in post-mitotic neurons it maintains steady-state H2A.Z occupancy to support transcription, dendritic arborization, and contextual fear memory (PMID:34407406); and in zebrafish it restricts precocious H2A.Z accumulation and premature activation of Sox-motif developmental genes before gastrulation (PMID:38159623, PMID:38187710). Beyond its chromatin role, a largely cytoplasmic ANP32E pool interacts with H2A.Z during G1 to govern H2A.Z protein stability, with H2A.Z stabilization dependent on phosphorylation and antagonized by PP2A (PMID:38482865, PMID:29524612). ANP32E also inhibits PP2A activity at cerebellar synapses during synaptogenesis in a phosphorylation-dependent manner (PMID:16420440). Excess ANP32E-driven H2A.Z turnover alters RNA polymerase II processivity and promotes R-loop accumulation and genomic instability at transcription-replication conflict sites (PMID:40382323).

Mechanistic history

Synthesis pass · year-by-year structured walk · 10 steps
  1. 2014 High

    Established that ANP32E is a dedicated H2A.Z-removing chaperone and defined the structural basis for its variant specificity, answering how a single factor discriminates H2A.Z from canonical H2A.

    Evidence Crystal structure of the ANP32E-ZID/H2A.Z/H2B complex with biochemical pulldowns and ChIP-seq in knockout cells; independently corroborated by in vitro binding and gain/loss-of-function ChIP-seq

    PMID:24463511 PMID:24613878

    Open questions at the time
    • Whether ANP32E acts catalytically or stoichiometrically in vivo not resolved
    • Recruitment determinants to specific genomic sites not defined
  2. 2015 High

    Showed ANP32E-mediated H2A.Z removal is functionally coupled to DNA damage response, defining a role in repair pathway choice.

    Evidence Live-cell imaging of DSB recruitment, ChIP at breaks, H4 acetylation assays, and repair pathway readouts in depleted cells

    PMID:26034280

    Open questions at the time
    • Signal recruiting ANP32E to breaks not identified
    • Direct link between H2A.Z removal and end-resection machinery not biochemically reconstituted
  3. 2020 High

    Demonstrated genome-wide that ANP32E restricts chromatin accessibility, establishing a hierarchical mechanism of H2A.Z accumulation around the TSS upon its loss.

    Evidence ATAC-seq, ChIP-seq, MNase-seq, and RNA-seq in knockout mouse fibroblasts

    PMID:33033242

    Open questions at the time
    • Causal order linking accessibility, TF binding, and transcription not fully dissected
    • Tissue-specificity of the effect not addressed
  4. 2021 High

    Distinguished steady-state from activity-dependent H2A.Z regulation in neurons and tied ANP32E to transcription, morphology, and memory.

    Evidence Conditional knockdown in hippocampal neurons with ChIP-seq, RNA-seq, dendritic morphology, and fear-conditioning behavior

    PMID:34407406

    Open questions at the time
    • Molecular basis for preference of steady-state over stimulus-induced removal unknown
    • Specific neuronal gene targets driving behavior not pinpointed
  5. 2022 High

    Showed ANP32E restrains developmental gene activation by limiting H2A.Z at Sox-motif promoters, extending its role to embryonic timing.

    Evidence Genetic knockout in zebrafish with ChIP-seq and RNA-seq

    PMID:38159623 PMID:38187710

    Open questions at the time
    • Why Sox-motif promoters are preferentially affected not explained
    • Connection to maternal-to-zygotic transition regulators not established
  6. 2018 Medium

    Linked ANP32E to PP2A-dependent regulation of H2A.Z protein stability and nuclear localization through phosphorylation.

    Evidence Yeast two-hybrid, H2A.Z S9A mutagenesis, PP2A inhibitor/overexpression rescue, MSK1 knockdown, and ChIP-seq

    PMID:29524612

    Open questions at the time
    • Direct kinase acting on H2A.Z S9 in this pathway not definitively established
    • Relationship between stability control and chromatin removal activity unclear
  7. 2006 Medium

    Identified an early, non-chromatin role for ANP32E (Cpd1) as a phosphorylation-dependent inhibitor of PP2A at synapses during synaptogenesis.

    Evidence Co-IP, PP2A activity assays, subcellular fractionation, electron microscopy, and ataxic mutant mouse models

    PMID:16420440

    Open questions at the time
    • Relationship between the membrane-bound 74/76 kDa form and the chromatin chaperone not reconciled
    • Direct synaptic substrates of inhibited PP2A not defined
  8. 2024 Medium

    Challenged the chromatin-remodeling-complex model by showing most ANP32E is cytoplasmic and engages H2A.Z in a G1-restricted, abundance-driven manner to control protein stability.

    Evidence Cell cycle synchronization, Co-IP, fractionation, mass spectrometry, and growth assays in U2OS cells

    PMID:38482865

    Open questions at the time
    • Conflicts with the stable p400-complex model and not yet reconciled
    • Mechanism coupling cytoplasmic stability control to nuclear H2A.Z eviction unknown
  9. 2025 Medium

    Connected ANP32E-driven H2A.Z turnover to RNA Pol II processivity, R-loop accumulation, and genomic instability in cancer.

    Evidence DRIP-seq, RNA Pol II ChIP-seq, ATR-inhibitor sensitivity, overexpression in breast cancer cells, and patient genomic analysis

    PMID:40382323

    Open questions at the time
    • Mechanistic link between H2A.Z turnover and R-loop induction is indirect
    • Whether MYC-ANP32E co-upregulation is causal or correlative not established
  10. 2026 Medium

    Defined a direct antagonism between ANP32E and the SRCAP subunit VPS72 at active promoters and reconstituted ANP32E's nucleosome-stabilizing activity.

    Evidence ChIP-seq, ATAC-seq, RNA-seq, genetic co-depletion epistasis, and in vitro nucleosome reconstitution (preprint)

    PMID:41646306

    Open questions at the time
    • Preprint, single lab, not yet peer-reviewed
    • Whether antagonism is direct competition for H2A.Z or independent opposing activities not fully resolved

Open questions

Synthesis pass · forward-looking unresolved questions
  • How ANP32E's cytoplasmic H2A.Z-stability function and its chromatin-eviction activity are mechanistically integrated, and how it is targeted to specific genomic loci, remains unresolved.
  • No unified model reconciling cytoplasmic vs chromatin roles
  • Locus-targeting determinants undefined
  • Stoichiometry/dynamics of removal vs deposition antagonism not quantified

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0042393 histone binding 3 GO:0098772 molecular function regulator activity 2 GO:0140096 catalytic activity, acting on a protein 1
Localization
GO:0005634 nucleus 2 GO:0005654 nucleoplasm 1 GO:0005829 cytosol 1
Pathway
R-HSA-4839726 Chromatin organization 3 R-HSA-74160 Gene expression (Transcription) 3 R-HSA-73894 DNA Repair 1
Complex memberships
p400/TIP60 H2A.Z histone-exchange complex

Evidence

Reading pass · 14 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2014 ANP32E is a histone chaperone that specifically removes H2A.Z from chromatin. It is a member of the p400/TIP60 H2A.Z histone-exchange complex and interacts with a short region of the docking domain of H2A.Z through a novel motif called the H2A.Z interacting domain (ZID). Crystal structure at 1.48 Å of the ANP32E-ZID/H2A.Z/H2B complex reveals that ANP32E stabilizes the H2A.Z/H2B dimer through a specific extension of the H2A.Z C-terminal α-helix. In ANP32E-knockout cells, H2A.Z shows genome-wide enrichment and accumulation at enhancers and insulators. Crystal structure (1.48 Å), Co-IP, co-fractionation, biochemical pulldown, chromatin immunoprecipitation followed by sequencing (ChIP-seq) in ANP32E-/- cells Nature High 24463511
2014 Anp32e preferentially associates with H2A.Z-H2B dimers over canonical H2A-H2B dimers both in vitro and in vivo. Crystal structure of the Anp32e chaperone domain (residues 186–232) bound to H2A.Z-H2B shows that residues 214–224 (absent in other Anp32 family members) specifically contact the extended H2A.Z αC helix. Overexpression of Anp32e causes global H2A.Z loss at +1 nucleosomes; depletion causes moderate global H2A.Z increase at +1 nucleosomes. Crystal structure, in vitro binding assays, genome-wide ChIP-seq, gain- and loss-of-function experiments Cell Research High 24613878
2015 Anp32e is rapidly recruited to DNA double-strand breaks (DSBs) and removes H2A.Z from nucleosomes, returning H2A.Z levels to basal within 10 min of DNA damage. H2A.Z removal by Anp32e disrupts inhibitory interactions between the histone H4 tail and the nucleosome surface, facilitating H4 acetylation. Loss of Anp32e leads to elevated nucleosomal H2A.Z, hypoacetylated chromatin at DSBs, increased CtIP-dependent end resection, ssDNA accumulation, and increased alternative non-homologous end joining repair. Live-cell imaging of DSB-recruited Anp32e, ChIP at DSBs, knockdown/knockout with specific repair pathway readouts, H4 acetylation assays Proceedings of the National Academy of Sciences of the United States of America High 26034280
2006 Anp32e/Cpd1 co-localizes with protein phosphatase 2A (PP2A) at synapses during cerebellar synaptogenesis. Synaptic Anp32e interacts with and inhibits PP2A activity. Phosphorylation of Anp32e is required for the Anp32e–PP2A interaction. A high-molecular-weight (74/76 kDa) membrane-bound form of Anp32e is expressed in a spatiotemporal pattern correlated with the cerebellar synaptogenesis period. Co-immunoprecipitation, PP2A activity assays, subcellular fractionation, electron microscopy, phosphorylation assays, ataxic mutant mouse models The European Journal of Neuroscience Medium 16420440
2018 ANP32E interacts with H2A.Z (C-terminus of ANP32E with N-terminus of H2A.Z, identified by yeast two-hybrid). ANP32E knockdown leads to proteasomal degradation and nuclear depletion of H2A.Z; this is reversed by PP2A inhibition and reproduced by PP2A catalytic subunit overexpression. ANP32E knockdown reduces H2A.Z phosphorylation; mutation of H2A.Z serine-9 abrogates its protein stability and nuclear localization. Nuclear mitogen and stress-induced kinase 1 (MSK1) knockdown phenocopies this effect. Yeast two-hybrid, knockdown, GFP-H2A.Z chimera, PP2A inhibitor treatment, site-directed mutagenesis (S9A), genome-wide ChIP-seq Biochimica et Biophysica Acta: Gene Regulatory Mechanisms Medium 29524612
2020 ANP32E antagonizes H2A.Z accumulation genome-wide to restrict chromatin accessibility in mouse fibroblasts. In the absence of ANP32E, H2A.Z accumulates at promoters in a hierarchical manner (first downstream, then upstream of the TSS), coinciding with improved nucleosome positioning, increased transcription factor binding, and elevated neighboring gene expression. ATAC-seq, ChIP-seq, ANP32E knockout mouse fibroblasts, MNase-seq, RNA-seq Nature Communications High 33033242
2021 In post-mitotic neurons, Anp32e regulates H2A.Z binding under steady-state conditions. Anp32e depletion causes H2A.Z-dependent impairment in transcription and dendritic arborization in cultured hippocampal neurons. In vivo, Anp32e depletion impairs recall of contextual fear memory and transcriptional regulation. Anp32e has lesser impact on stimulus-induced (activity-dependent) H2A.Z removal compared to steady-state regulation. Conditional knockdown in hippocampal neurons, ChIP-seq, RNA-seq, fear-conditioning behavioral assay, dendritic morphology analysis Cell Reports High 34407406
2022 In zebrafish, Anp32e restricts H2A.Z accumulation specifically at Sox motif-containing promoters. Genetic removal of Anp32e leads to precocious H2A.Z accumulation and premature transcriptional activation of Sox motif-associated developmental genes before gastrulation. Genetic knockout of anp32e in zebrafish, ChIP-seq, RNA-seq Developmental Biology High 38159623 38187710
2022 Anp32e promotes renal interstitial fibrosis by upregulating TGF-β1 and p-Smad3, leading to deposition of fibronectin and collagen type I. Overexpression of Anp32e alone (without TGF-β1 stimulation) induces fibrosis-related protein deposition; this is reversed by the TGF-β1 inhibitor SB431542, placing Anp32e upstream of the TGF-β1/Smad3 pathway. Gain- and loss-of-function in BUMPT cells and UUO mouse model, western blot, TGF-β1 inhibitor rescue assay International Journal of Biological Sciences Medium 36263179
2024 ANP32E preferentially interacts with H2A.Z during the G1 phase of the cell cycle in both cytoplasm and nucleoplasm of human U2OS cells. Most ANP32E is cytoplasmic, not chromatin-associated, and is not a stable component of the p400 remodeling complex. In the cytoplasm, ANP32E interacts with H2A.Z in G1 in response to increased H2A.Z protein abundance and regulates H2A.Z protein stability. This challenges the model that ANP32E removes H2A.Z from chromatin as part of a remodeling complex. Cell cycle synchronization, Co-IP, subcellular fractionation, mass spectrometry, U2OS cell knockdown with growth assays Molecular and Cellular Biology Medium 38482865
2025 ANP32E-driven H2A.Z turnover alters RNA polymerase II processivity, leading to accumulation of long R-loops at transcription-replication conflict (TRC) sites. ANP32E overexpression enhances TRC formation and activates ATR-dependent DNA damage response, predisposing cancer cells to R-loop-mediated genomic fragility. Tumors with co-upregulation of MYC and ANP32E show increased genomic instability. Genome-wide R-loop mapping (DRIP-seq), RNA Pol II ChIP-seq, ATR inhibitor sensitivity assays, ANP32E overexpression in breast cancer cells, patient genomic data analysis Nature Communications Medium 40382323
2026 ANP32E and the SRCAP subunit VPS72 co-occupy active gene promoters and function antagonistically on H2A.Z. VPS72 promotes H2A.Z incorporation, acetylation, BRG1 recruitment, and transcription; ANP32E constrains these features by stabilizing nucleosomes. Loss of ANP32E increases VPS72 binding, chromatin accessibility, and transcription; co-depletion of VPS72 reverses these effects. In vitro reconstitution assays demonstrate that ANP32E promotes nucleosome assembly and prevents DNA unwrapping. ChIP-seq, ATAC-seq, RNA-seq, genetic co-depletion epistasis, in vitro nucleosome reconstitution assays Research Square (preprint)preprint Medium 41646306
2018 ANP32E promotes G1/S cell cycle transition in triple-negative breast cancer cells by transcriptionally inducing E2F1. ANP32E inhibition suppresses tumor formation in vivo. siRNA knockdown, overexpression, cell cycle analysis, E2F1/cyclin E expression analysis, xenograft mouse model Molecular Oncology Low 29633513
2024 ANP32E regulates esophageal cancer progression and ferroptosis via the p53/SLC7A11 axis. ANP32E depletion increases p53 expression; p53 inhibition partially reverses the suppressed proliferation and enhanced ferroptosis in ANP32E-depleted cells. ANP32E knockout cell models, xenograft, RNA-seq, ferroptosis inhibitor rescue (ferrostatin-1), p53 inhibitor rescue, western blot International Immunopharmacology Low 39566382

Source papers

Stage 0 corpus · 30 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2014 ANP32E is a histone chaperone that removes H2A.Z from chromatin. Nature 221 24463511
2015 Histone chaperone Anp32e removes H2A.Z from DNA double-strand breaks and promotes nucleosome reorganization and DNA repair. Proceedings of the National Academy of Sciences of the United States of America 119 26034280
2014 Anp32e, a higher eukaryotic histone chaperone directs preferential recognition for H2A.Z. Cell research 112 24613878
2017 Downregulation of miRNA-141 in breast cancer cells is associated with cell migration and invasion: involvement of ANP32E targeting. Cancer medicine 79 28220627
2018 ANP32E induces tumorigenesis of triple-negative breast cancer cells by upregulating E2F1. Molecular oncology 51 29633513
2020 Genome-wide chromatin accessibility is restricted by ANP32E. Nature communications 43 33033242
2003 Anp32e (Cpd1) and related protein phosphatase 2 inhibitors. Cerebellum (London, England) 37 14964690
2019 ANP32E, a Protein Involved in Steroid-Refractoriness in Ulcerative Colitis, Identified by a Systems Biology Approach. Journal of Crohn's & colitis 36 30329026
2006 Anp32e/Cpd1 regulates protein phosphatase 2A activity at synapses during synaptogenesis. The European journal of neuroscience 27 16420440
2020 Up-regulated ANP32E promotes the thyroid carcinoma cell proliferation and migration via activating AKT/mTOR/HK2-mediated glycolysis. Gene 25 32304784
2010 Generation and characterization of the Anp32e-deficient mouse. PloS one 25 21049064
2021 Long non-coding RNA NORAD promotes pancreatic cancer stem cell proliferation and self-renewal by blocking microRNA-202-5p-mediated ANP32E inhibition. Journal of translational medicine 23 34551785
2022 Lockd promotes myoblast proliferation and muscle regeneration via binding with DHX36 to facilitate 5' UTR rG4 unwinding and Anp32e translation. Cell reports 20 35675771
2021 The histone chaperone Anp32e regulates memory formation, transcription, and dendritic morphology by regulating steady-state H2A.Z binding in neurons. Cell reports 19 34407406
2020 Hsa-let-7c exerts an anti-tumor function by negatively regulating ANP32E in lung adenocarcinoma. Tissue & cell 15 32746998
2024 ANP32E promotes esophageal cancer progression and paclitaxel resistance via P53/SLC7A11 axis-regulated ferroptosis. International immunopharmacology 14 39566382
2022 ANP32E contributes to gastric cancer progression via NUF2 upregulation. Molecular medicine reports 13 35795988
2022 Anp32e promotes renal interstitial fibrosis by upregulating the expression of fibrosis-related proteins. International journal of biological sciences 12 36263179
2018 Transcriptional regulation mediated by H2A.Z via ANP32e-dependent inhibition of protein phosphatase 2A. Biochimica et biophysica acta. Gene regulatory mechanisms 12 29524612
2021 Subtype-Independent ANP32E Reduction During Breast Cancer Progression in Accordance with Chromatin Relaxation. BMC cancer 8 34922480
2002 Molecular cloning and characterization of a novel human gene (ANP32E alias LANPL) from human fetal brain. Cytogenetic and genome research 8 12438741
2023 Anp32e protects against accumulation of H2A.Z at Sox motif containing promoters during zebrafish gastrulation. Developmental biology 6 38159623
2024 ANP32e Binds Histone H2A.Z in a Cell Cycle-Dependent Manner and Regulates Its Protein Stability in the Cytoplasm. Molecular and cellular biology 5 38482865
2025 ANP32E drives vulnerability to ATR inhibitors by inducing R-loops-dependent transcription replication conflicts in triple negative breast cancer. Nature communications 4 40382323
2013 Targeted ANP32E mutant mice do not demonstrate obvious movement defects. PloS one 4 23675506
2024 FOXCUT regulates the malignant phenotype of triple-negative breast Cancer via the miR-337-3p/ANP32E Axis. Genomics 3 38944356
2024 Knockdown of ANP32E inhibits colorectal cancer cell growth and glycolysis by regulating the AKT/mTOR pathway. Open life sciences 2 38585643
2025 Inhibiting ANP32E protects against acute kidney injury by regulating autophagy via the AMPK pathway. iScience 1 40686617
2026 Cooperative and Opposing Functions of ANP32E and VPS72 Govern Gene Promoter Chromatin Status. Research square 0 41646306
2023 Anp32e protects against accumulation of H2A.Z at Sox motif containing promoters during zebrafish gastrulation. bioRxiv : the preprint server for biology 0 38187710

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