Affinage

FANCD2

Fanconi anemia group D2 protein · UniProt Q9BXW9

Length
1451 aa
Mass
164.1 kDa
Annotated
2026-04-28
100 papers in source corpus 49 papers cited in narrative 50 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

FANCD2 is the central effector of the Fanconi anemia (FA) DNA repair pathway, functioning as a monoubiquitin-regulated DNA clamp that protects stalled replication forks and coordinates interstrand crosslink (ICL) repair, homologous recombination, and R-loop suppression. Upon replication stress, ATR phosphorylates FANCI within the FANCI–FANCD2 (ID2) heterodimer, priming a conformational change that enables the FA core complex (FANCB–FANCL–FAAP100 catalytic module with UBE2T as E2) to monoubiquitinate FANCD2 at K561; this modification induces the ID2 complex to clamp onto dsDNA, slide along it, and stall specifically at ss–dsDNA junctions characteristic of stalled forks, where it recruits downstream nucleases (FAN1, XPF-ERCC1/SLX4), recombination factors (CtIP, BRCA2, RAD51), translesion polymerases (REV1, pol η), and RNA-processing factors (hnRNPU, DDX47, SRSF1) to execute repair, fork restart, and mRNA export (PMID:39085614, PMID:20603015, PMID:24726325, PMID:24794434, PMID:30431240, PMID:38165804). The monoubiquitin mark is removed by USP1–UAF1 in a DNA- and UAF1-dependent manner, and cycling between ubiquitinated and deubiquitinated states is essential for ICL repair, hematopoietic stem cell maintenance, and tumor suppression via TAp63-mediated senescence (PMID:18082605, PMID:31253762, PMID:20506303, PMID:23806336). Biallelic loss-of-function mutations in FANCD2 cause Fanconi anemia complementation group D2, characterized by bone marrow failure, chromosomal instability, and cancer predisposition (PMID:11239453).

Mechanistic history

Synthesis pass · year-by-year structured walk · 19 steps
  1. 2001 High

    Positional cloning identified FANCD2 as the gene mutated in FA complementation group D2, establishing it as a novel nuclear protein required for resistance to DNA crosslinking agents.

    Evidence Positional cloning and retroviral complementation of FA-D2 patient cells

    PMID:11239453

    Open questions at the time
    • No biochemical activity assigned
    • Mechanism of crosslinker sensitivity unknown
  2. 2002 High

    Discovery that FANCD2 is monoubiquitinated at K561 during S phase and after DNA damage, and that this modification directs it to BRCA1/RAD51 nuclear foci, revealed the key regulatory switch in the FA pathway.

    Evidence Immunofluorescence co-localization, K561R mutagenesis, chromatin fractionation in human cells

    PMID:12239151

    Open questions at the time
    • Identity of the E3 ligase unknown
    • Whether monoubiquitination is sufficient for chromatin binding unresolved
  3. 2002 High

    FANCD2 was placed in the ATM/NBS1 S-phase checkpoint axis through its interaction with NBS1 and ATM-dependent phosphorylation, broadening FANCD2's role beyond ICL repair to general replication stress signaling.

    Evidence Reciprocal co-IP of FANCD2–NBS1, S-phase checkpoint assay in ATM-deficient cells

    PMID:12447395

    Open questions at the time
    • Precise phosphorylation sites on FANCD2 by ATM not mapped
    • Relationship between phosphorylation and monoubiquitination unclear
  4. 2004 High

    ATR kinase and RPA were shown to be upstream regulators required for FANCD2 monoubiquitination and foci formation, establishing ATR as the primary kinase linking replication stress sensing to FA pathway activation.

    Evidence siRNA and Seckel syndrome cells with chromosome analysis and immunofluorescence

    PMID:14988723 PMID:15314022

    Open questions at the time
    • Whether ATR directly phosphorylates FANCD2 or acts indirectly via FANCI not resolved at this stage
  5. 2004 High

    Monoubiquitination at K561 was shown to be required for chromatin binding, and FANCD2 was linked to BRCA2 through direct interaction, placing FANCD2 at the interface between the FA pathway and homologous recombination.

    Evidence Chromatin fractionation of K561R mutant in FANCD2−/− fibroblasts; yeast two-hybrid and co-IP for BRCA2 interaction

    PMID:15115758 PMID:15454491

    Open questions at the time
    • How monoubiquitination promotes chromatin binding mechanistically unknown
    • Whether BRCA2 interaction requires monoubiquitination untested
  6. 2007 High

    Discovery of FANCI as a FANCD2 paralog forming the ID2 complex, with a reciprocal ubiquitin-locking mechanism where each partner's ubiquitination stabilizes the other's, revealed the fundamental heterodimeric unit of the pathway.

    Evidence Mass spectrometry identification, co-IP, chromatin fractionation, mutual ubiquitination dependence assays

    PMID:17412408

    Open questions at the time
    • Structural basis for reciprocal locking unknown
    • Whether the complex binds DNA before or after ubiquitination unresolved
  7. 2007 High

    USP1 was identified as the deubiquitinase that removes monoubiquitin from FANCD2, and its disruption showed that constitutive monoubiquitination paradoxically impairs ICL repair, demonstrating that cycling of the ubiquitin mark is essential.

    Evidence USP1 gene disruption in DT40 cells, chromatin fractionation, MMC sensitivity

    PMID:18082605

    Open questions at the time
    • How USP1 recognizes FANCD2 substrate specificity unknown
    • Role of UAF1 cofactor not yet defined
  8. 2010 High

    Identification of FAN1 as a nuclease recruited by monoubiquitinated FANCD2 provided the first direct mechanistic link between FANCD2 activation and enzymatic processing of ICL lesions.

    Evidence Co-IP of FAN1 with FANCD2-Ub, in vitro nuclease assays, siRNA with ICL sensitivity

    PMID:20603015

    Open questions at the time
    • Whether FAN1 is the sole incision nuclease or acts redundantly with XPF-ERCC1 unclear
  9. 2011 High

    The 3.4 Å crystal structure of the FANCI–FANCD2 complex revealed that ubiquitination and phosphorylation sites map to the I–D interface and that both subunits harbor DNA-binding sites, providing a structural framework for understanding pathway activation.

    Evidence X-ray crystallography of the ~300 kDa ID complex, in vitro DNA binding

    PMID:21764741

    Open questions at the time
    • Structure was of unmodified complex; ubiquitinated state unsolved
    • How DNA binding triggers ubiquitination structurally unclear
  10. 2012 High

    Biochemical reconstitution showed that DNA — particularly 5′-flap structures mimicking stalled forks — robustly stimulates FANCD2 monoubiquitination in the ID complex through FANCI's DNA-binding activity, explaining the DNA damage dependence of pathway activation.

    Evidence In vitro ubiquitylation with purified proteins, various DNA substrates, FANCI DNA-binding mutants

    PMID:22287633

    Open questions at the time
    • Which FA core complex subunits contribute to DNA-stimulated activity not fully resolved
  11. 2013 High

    Monoubiquitinated FANCD2 was shown to have a tumor-suppressive function by activating TAp63 transcription to promote senescence, and FANCD2 was found necessary for hematopoietic stem cell maintenance, extending its biological role beyond DNA repair.

    Evidence Fancd2−/− and Usp1−/− mouse cancer models; HSC transplantation and flow cytometry in Fancd2−/− mice

    PMID:20506303 PMID:23806336

    Open questions at the time
    • How FANCD2 activates TAp63 transcription mechanistically unknown
    • Whether HSC defect is solely due to DNA repair impairment unresolved
  12. 2014 High

    FANCD2-dependent recruitment of XPF-ERCC1/SLX4 for ICL unhooking and CtIP for HR channeling placed monoubiquitinated FANCD2 as a platform that coordinates sequential repair steps at crosslinks and stalled forks.

    Evidence Xenopus egg extract ICL repair with immunodepletion; co-IP and HR/NHEJ reporter assays for CtIP

    PMID:24556218 PMID:24726325 PMID:24794434

    Open questions at the time
    • Order of recruitment of downstream nucleases versus recombination factors not fully established
    • Whether CtIP interaction requires monoubiquitination directly debated
  13. 2014 High

    Purification and reconstitution of the native FA core complex showed that FANCL embedded within a FANCB–FANCL–FAAP100 module is the catalytic E3 for FANCD2 monoubiquitination, resolving years of uncertainty about the enzymatic source.

    Evidence Purified avian FA core complex, in vitro ubiquitination with genetic dissection of minimal subcomplex

    PMID:24905007

    Open questions at the time
    • How the full 8-subunit core complex enhances specificity beyond the minimal module unclear
  14. 2017 Medium

    FANCD2 was found to recruit RNA-processing factors (hnRNPU, DDX47) and spliceosomal protein SF3B1 to chromatin, suppressing R-loop accumulation at fragile genes during replication stress, revealing a non-repair function in RNA metabolism.

    Evidence Co-IP/MS, ChIP-seq at large fragile genes, PLA, R-loop detection (DRIP)

    PMID:29030393 PMID:30431240

    Open questions at the time
    • Whether R-loop suppression is direct or secondary to fork stabilization unclear
    • FANCD2 enzymatic contribution to RNA processing not defined
  15. 2019 High

    The allosteric mechanism of FANCD2 monoubiquitination was resolved: FANCL rewires UBE2T's active site through a basic triad to engage an acidic patch near K561, explaining site specificity; separately, USP1-UAF1 deubiquitination was shown to require DNA binding by UAF1/RAD51AP1.

    Evidence In vitro ubiquitination plus NMR for allosteric mechanism; reconstituted deubiquitination with UAF1 DNA-binding mutants

    PMID:31253762 PMID:31873223

    Open questions at the time
    • Complete structural view of UBE2T–FANCL–ID2 ternary complex not available
    • How DNA binding by UAF1 couples to USP1 catalysis structurally unresolved
  16. 2020 High

    Monoubiquitination of FANCD2 was shown to induce a large-scale conformational change that clamps the ID2 complex onto dsDNA, forming filament-like arrays; FANCI ubiquitination protects the FANCD2-Ub mark from USP1, establishing the molecular logic of the dual-lock mechanism.

    Evidence In vitro reconstitution with EM visualization; deubiquitination protection assays; DNA binding measurements

    PMID:32167469 PMID:32510829

    Open questions at the time
    • Filament function in vivo not established
    • Whether clamping alone suffices for downstream effector recruitment unknown
  17. 2020 High

    ATR was shown to directly phosphorylate FANCI at S556/S559/S565 to stabilize the FANCI–DNA–FANCD2 complex and inhibit USP1-mediated deubiquitination, completing the signaling circuit from damage sensing to clamp activation.

    Evidence Reconstituted ATR kinase assay, phosphomimetic/phosphodead FANCI mutants, in vitro ubiquitination/deubiquitination

    PMID:32117957

    Open questions at the time
    • Whether other kinases contribute in vivo remains open
    • Dephosphorylation step that resets the cycle not identified
  18. 2022 High

    Cryo-EM structures of phosphomimetic FANCI-containing ID2 complexes showed that ATR-mediated phosphorylation destabilizes the open state and drives clamp closure around DNA independently of the FA core complex, providing structural proof for the phosphorylation-priming model.

    Evidence Cryo-EM of phosphomimetic FANCI–FANCD2–DNA complexes, conformational dynamics analysis

    PMID:36050501

    Open questions at the time
    • Transition from closed/phosphorylated to ubiquitinated state not captured structurally
    • Role of DNA sequence or structure in closure not systematically tested
  19. 2024 High

    Single-molecule imaging demonstrated that ubiquitinated ID2 slides on dsDNA and stalls specifically at ss–dsDNA junctions, with cryo-EM revealing junction-specific contacts distinct from the sliding state, providing a unified mechanism for how FANCD2 surveys and protects stalled replication forks.

    Evidence Single-molecule sliding assays, cryo-EM structures of sliding and stalled states on junction DNA

    PMID:39085614

    Open questions at the time
    • In vivo validation of sliding clamp behavior at endogenous forks not yet achieved
    • Whether junction recognition triggers downstream effector handoff unknown

Open questions

Synthesis pass · forward-looking unresolved questions
  • Key open questions include how ubiquitinated FANCD2 discriminates among and sequentially recruits its numerous downstream effectors, the structural basis of the full FA core complex–ID2–DNA assembly, and how the non-repair functions of FANCD2 (R-loop suppression, mRNA export, TAp63 activation) are coordinated with its canonical DNA repair role.
  • Complete structural model of FA core complex–ID2–DNA ternary complex lacking
  • Mechanism of effector handoff at stalled forks unresolved
  • Phosphatase that reverses FANCI phosphorylation not identified
  • Whether mitochondrial FANCD2 function is physiologically significant remains uncertain

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0003677 DNA binding 5 GO:0060090 molecular adaptor activity 4
Localization
GO:0005634 nucleus 4 GO:0005694 chromosome 4
Pathway
R-HSA-73894 DNA Repair 6 R-HSA-69306 DNA Replication 5 R-HSA-1640170 Cell Cycle 3 R-HSA-8953854 Metabolism of RNA 3
Partners
BRCA1BRCA2CTIPFAN1FANCIFANCLNBS1USP1
Complex memberships
FANCI-FANCD2 (ID2) complex

Evidence

Reading pass · 50 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2001 FANCD2 is a novel 1451 amino acid nuclear protein encoded by a gene with 44 exons; retroviral transduction of FANCD2 cDNA into FA-D2 cells complemented MMC sensitivity, establishing it as the FA complementation group D2 gene product. Positional cloning, retroviral complementation assay Molecular cell High 11239453
2002 FANCD2 undergoes monoubiquitination at K561 during S phase and in response to DNA damage, and monoubiquitinated FANCD2 colocalizes with BRCA1 and RAD51 in S-phase nuclear foci; monoubiquitination is required for normal cell-cycle progression following MMC exposure. Immunofluorescence co-localization, cell fractionation, mutational analysis (K561R) Blood High 12239151
2002 FANCD2 interacts with NBS1; ionizing radiation activates ATM-dependent, NBS1-dependent phosphorylation of FANCD2 to mediate an S-phase checkpoint, while MMC activates colocalization of FANCD2 and NBS1 in subnuclear foci. Co-immunoprecipitation, immunofluorescence, S-phase checkpoint assay Nature cell biology High 12447395
2003 The purified BRCA1/BARD1 complex reconstitutes monoubiquitination of FANCD2 in vitro with E1 and UbcH5a; however, BRCA1/BARD1 E3 ligase activity is not essential for in vivo FANCD2 monoubiquitination, whereas BRCA1 is required for FANCD2 targeting to DNA damage sites. In vitro ubiquitination reconstitution, siRNA knockdown, RING domain mutagenesis in chicken DT40 cells Molecular cell High 12887909
2004 ATR kinase and RPA1 are required for efficient FANCD2 monoubiquitination and assembly of FANCD2 nuclear foci; deficiency of ATR results in radial chromosomes upon MMC treatment, mimicking FA chromosome instability. siRNA silencing, cell line analysis (Seckel syndrome), immunofluorescence, chromosome analysis Genes & development High 15314022
2004 FANCD2 directly interacts with BRCA2 at a conserved C-terminal site; FANCD2 and BRCA2 co-immunoprecipitate from human and hamster cell extracts; FANCD2 colocalizes with RAD51 and PCNA following replication fork stalling, suggesting a role in repair of replication-associated DSBs. Yeast two-hybrid, co-immunoprecipitation, immunofluorescence co-localization Human molecular genetics High 15115758
2004 Monoubiquitination of FANCD2 at K561 is required for chromatin binding; the C-terminal residue D1428 encoded by exon 44 is independently required for functional complementation; a FANCD2-K561R mutant fails to bind chromatin. Stable transfection of FANCD2 mutants in FANCD2-/- fibroblasts, chromatin fractionation, MMC sensitivity assay Blood High 15454491
2004 ICL-induced S-phase checkpoint requires ATR kinase; FANCD2 is phosphorylated in an ATR-dependent manner and ATR colocalizes with FANCD2; the checkpoint involves parallel branches: CHK1 and FANCs/NBS1 acting downstream of ATR. siRNA, immunofluorescence, S-phase checkpoint assays, kinase inhibition The EMBO journal High 14988723
2007 FANCI is a paralog of FANCD2 that associates with FANCD2 to form the ID complex on chromatin in response to DNA damage; FANCI is monoubiquitinated, and ubiquitination of each protein (FANCI and FANCD2) is required to maintain ubiquitin on the other, revealing a dual ubiquitin-locking mechanism. Mass spectrometry identification, co-immunoprecipitation, chromatin fractionation, monoubiquitination assays Cell High 17412408
2007 USP1 deubiquitinase removes monoubiquitin from FANCD2; USP1 disruption causes constitutively chromatin-bound monoubiquitinated FANCD2 and crosslinker sensitivity, demonstrating that FANCD2 deubiquitination is required for efficient DNA crosslink repair. USP1 gene disruption in chicken DT40 cells, chromatin fractionation, MMC sensitivity assay Molecular cell High 18082605
2008 FANCG promotes formation of a complex containing BRCA2, FANCD2, and XRCC3; phosphorylation of FANCG serine 7 is required for co-precipitation of BRCA2, XRCC3, and FANCD2; FANCG and XRCC3 are epistatic for sensitivity to crosslinking agents. Co-immunoprecipitation, phosphorylation site mutagenesis, DT40 genetic epistasis Oncogene Medium 18212739
2009 Monoubiquitinated FANCD2 colocalizes with telomeres and PML bodies in ALT cells; FA core complex components FANCA and FANCL regulate FANCD2 monoubiquitination and its telomeric localization; FANCD2 depletion causes loss of detectable telomeres and decreased T-SCE in ALT cells. siRNA knockdown, immunofluorescence co-localization, T-SCE assay Nucleic acids research Medium 19129235
2009 FANCM chromatin binding and DNA damage-induced phosphorylation are partially regulated by the downstream FA pathway protein FANCD2 in Xenopus egg extracts. Xenopus egg extract system, immunodepletion, chromatin fractionation The Journal of biological chemistry Medium 19633289
2010 FAN1 (KIAA1018) interacts with monoubiquitinated FANCD2 and is recruited to sites of DNA damage by monoubiquitinated FANCD2; FAN1 has 5' flap endonuclease and 5' exonuclease activities mediated by a VRR_nuc domain; FAN1 depletion causes ICL hypersensitivity. Co-immunoprecipitation, in vitro nuclease assay, siRNA depletion, ICL repair assay Cell High 20603015
2010 RAD18 E3 ubiquitin ligase binds FANCD2 and is required for efficient monoubiquitylation and chromatin localization of both FANCD2 and FANCI; the RING domain of RAD18 is required for this interaction and chromatin loading. Co-immunoprecipitation, RAD18 knockout cells, chromatin fractionation, RING domain mutagenesis Blood Medium 21355096
2010 Fancd2-deficient mouse bone marrow shows decreased HSC frequency (Lin-Sca-1+Kit+ and SLAM marker populations) and defective long-term in vivo repopulating ability, establishing FANCD2 as required for hematopoietic stem cell maintenance. Fancd2-/- mouse model, flow cytometry, cobblestone area-forming cell assay, transplantation Stem cells High 20506303
2011 Crystal structure of the ~300 kDa FANCI-FANCD2 (ID) complex at 3.4 Å reveals that monoubiquitination and regulatory phosphorylation sites map to the I-D interface; both proteins have binding sites for single- and double-stranded DNA, suggesting the ID complex recognizes DNA structures at stalled replication forks. X-ray crystallography, electron density mapping of FANCI-DNA crystals, in vitro DNA binding Science High 21764741
2012 FANCD2 contains a CUE ubiquitin-binding domain that mediates noncovalent ubiquitin binding in vitro; the CUE domain is required for interaction with FANCI, chromatin retention of monoubiquitinated FANCD2 and FANCI, and efficient ICL repair. CUE domain mutagenesis, in vitro ubiquitin binding assay, co-immunoprecipitation, ICL repair assay Blood Medium 22855611
2012 DNA robustly stimulates FANCD2 monoubiquitylation in the FANCI-FANCD2 complex in vitro; this stimulation strictly requires FANCI and FANCI's DNA binding activity; 5' flapped DNA (mimicking arrested replication forks) achieves ~70% monoubiquitylation in vitro. In vitro ubiquitylation reconstitution with purified proteins and various DNA substrates, FANCI DNA binding mutants Nucleic acids research High 22287633
2012 DNA damage-induced FA pathway activation triggers dissociation of FANCD2 from FANCI; FANCI phosphorylation is the molecular trigger for dissociation; FANCD2 monoubiquitination significantly precedes FANCI monoubiquitination; FANCD2 binds replicating chromatin prior to and independently of FANCI. Co-immunoprecipitation, chromatin fractionation, phosphomimetic/phosphodead FANCI mutants Nucleic acids research Medium 22753026
2013 Monoubiquitinated FANCD2 (FANCD2-Ub) activates transcription of the tumor suppressor TAp63 to promote cellular senescence and block skin tumorigenesis; Usp1-deficient mice with elevated FANCD2-Ub are resistant to skin tumors while Fancd2-deficient mice are susceptible. Fancd2-/- and Usp1-/- mouse models, Ras-driven skin carcinogenesis, TAp63 transcription assay Molecular cell High 23806336
2013 mTOR regulates FANCD2 expression through NF-κB; mTOR loss increases NF-κB nuclear translocation and NF-κB binding to the FANCD2 promoter, suppressing FANCD2 expression; exogenous FANCD2 rescues the DDR defect caused by mTOR inhibition. mTOR gene targeting in HSPCs, NF-κB ChIP at FANCD2 promoter, rescue by FANCD2 re-expression Leukemia Medium 23538752
2014 XPF-ERCC1 cooperates with SLX4/FANCP to carry out DNA unhooking incisions during ICL repair in Xenopus egg extracts; efficient recruitment of XPF-ERCC1 and SLX4 to ICLs depends on FANCD2 and its monoubiquitination. Xenopus egg extract ICL repair system, immunodepletion, site-specific ICL substrates Molecular cell High 24726325
2014 The genetic and biochemical basis of FANCD2 monoubiquitination: purified native FA core complex reconstitutes FANCD2 monoubiquitination in vitro; FANCL must be embedded in the complex for maximal activity and site specificity; a minimal FANCB-FANCL-FAAP100 subcomplex functions as the monoubiquitination module. Purification of native avian FA core complex, in vitro ubiquitination reconstitution, genetic dissection of minimal complex Molecular cell High 24905007
2014 Monoubiquitinated FANCD2 tethers CtIP to damaged chromatin via physical interaction; this channels ICL-generated DSBs into HR; CtIP mutants defective in FANCD2 binding fail to associate with damaged chromatin and show increased NHEJ and ICL hypersensitivity. Co-immunoprecipitation, chromatin fractionation, ICL sensitivity assay, HR/NHEJ reporter assays Cell reports Medium 24794434
2014 CtIP is a novel interaction partner of FANCD2; CtIP binds and stabilizes FANCD2 in a DNA damage- and FA core complex-independent manner; FANCD2 recruits CtIP to stalled replication forks to promote fork restart and suppress new origin firing, dependent on BRCA1. Co-immunoprecipitation, DNA fiber assay, chromatin fractionation, siRNA knockdown Human molecular genetics Medium 24556218
2015 FANCI, but not FANCD2, is needed for efficient FA core complex foci formation; FANCI functions upstream of FA core complex recruitment independently of FANCD2; FANCD2 monoubiquitination is not required for this FANCI function, but USP1-mediated FANCI deubiquitination is. siRNA, FANCI/FANCD2 knockout cell lines, immunofluorescence foci assay PLoS genetics Medium 26430909
2015 FANCD2 cooperates with BRCA2 and RAD51 to protect stalled replication forks from nucleolytic degradation and recruits BLM helicase to promote replication fork restart while suppressing new origin firing. DNA fiber assay, siRNA knockdown, co-immunoprecipitation, replication restart assays Human molecular genetics Medium 24556218
2015 REV1 is recruited to DNA damage sites via its ubiquitin-binding motifs and monoubiquitinated FANCD2, acting downstream of RAD18; FANCD2 and REV1 are epistatic for camptothecin sensitivity; REV1 protects nascent replication tracts from degradation by stabilizing RAD51 filaments. Laser stripe recruitment assay, FANCD2-Ub chimeric protein expression, DNA fiber assay, epistasis analysis Nucleic acids research Medium 26187992
2016 FANCD2-FANCI complex is recruited to stalled replication forks before monoubiquitination; cryo-EM structure of the human complex reveals an inner cavity large enough for dsDNA and a Tower domain harboring disease-causing mutations; fork recruitment triggers the activating monoubiquitination. Cryo-EM structure determination, disease mutation mapping, reconstitution assays Nature communications High 27405460
2016 FANCD2 is required for fork protection and fork restart in BRCA1/2-deficient tumors; FANCD2 promotes Polθ recruitment at damage sites and alt-EJ repair; loss of FANCD2 in BRCA1/2-deficient tumors enhances cell death, revealing a synthetic lethal relationship. DNA fiber assay, siRNA in BRCA1/2-deficient cell lines, alt-EJ reporter, xenograft tumor model Cell reports High 27264184
2016 Monoubiquitinated FANCD2 acts in opposition to BLM helicase at ALT telomeres; FANCD2 depletion causes hyper-ALT phenotype including increased extrachromosomal telomeric repeats; increases are suppressed by BLM but not RAD51 loss. siRNA depletion, FANCD2/BLM double knockdown, telomere FISH, C-circle assay Human molecular genetics Medium 27427384
2017 FANCD2 has two major DNA binding domains consisting of conserved lysine residues; an N-terminal domain also contains nuclear localization sequences; mutations in the bifunctional DNA binding/NLS domain reduce FANCD2 monoubiquitination and increase MMC sensitivity. Synthetic peptide mapping, electromobility shift assay, NLS mutagenesis, complementation assay Nucleic acids research Medium 28666371
2017 FANCD2 interacts with RNA processing factors hnRNP U and DDX47; FANCD2 recruits these factors to chromatin at large fragile genes and promotes efficient processing of long RNA transcripts, thereby suppressing R-loop levels during mild replication stress. Co-immunoprecipitation/mass spectrometry, ChIP-seq, PLA, siRNA knockdown, R-loop detection The FEBS journal Medium 30431240
2017 FANCD2 functionally impacts mitochondrial ATP production through interaction with ATP5α; monoubiquitinated FANCD2 is required for this interaction and for proper mitochondrial localization of ATP5α; non-monoubiquitinated FANCD2 (K561R) fails to interact with ATP5α and cells show reduced mitochondrial ATP production. Co-immunoprecipitation, mitochondrial fractionation, ATP production assay, protein docking Scientific reports Low 28687786
2017 FANCI and FANCD2 associate with the spliceosomal protein SF3B1; DNA replication stress induces ATR- and FANCI-dependent release of SF3B1 from nuclear speckles; both FANCD2 and FANCI prevent accumulation of postcatalytic intron lariats in chromatin. Co-immunoprecipitation, proximity ligation assay, immunofluorescence, lariat RT-PCR The Journal of cell biology Medium 29030393
2018 FANCD2 accumulates at the central regions of large fragile genes during replication stress in an R-loop-dependent manner; FANCD2 monoubiquitination and RPA foci formation are still induced after R-loop depletion; increased FANCD2-R-loop proximity is detected by PLA following aphidicolin treatment. ChIP-seq, PLA, siRNA depletion of R-loops (RNaseH1 overexpression), immunofluorescence Nucleic acids research Medium 29394375
2019 FANCL allosterically activates UBE2T (E2) to drive site-specific FANCD2 monoubiquitination; FANCL rewires the intraresidue network of UBE2T to engage an acidic patch near K561 on FANCD2 through a basic triad unique to UBE2T. In vitro ubiquitination reconstitution, NMR, mutagenesis of UBE2T active site, site-specificity assays Nature chemical biology High 31873223
2019 Efficient FANCD2 deubiquitination by USP1-UAF1 requires DNA and DNA-binding by UAF1; RAD51AP1's DNA binding activity can substitute for UAF1 DNA binding in FANCD2 deubiquitination; UAF1 and RAD51AP1 DNA binding is important for FANCD2 deubiquitination in cells. Biochemical reconstitution with purified proteins, UAF1/RAD51AP1 mutants, cellular deubiquitination assays Nature communications High 31253762
2019 Nuclear receptors COUP-TFII and TR4 directly interact with FANCD2 and form a complex that recruits MUS81 endonuclease and PCNA-POLD3 replication complex to ALT telomeres; this noncanonical FANCD2 pathway operates independently of the FA core complex or FANCD2 monoubiquitination. Co-immunoprecipitation, ChIP at telomeres, gene knockdown, telomere function assays Science advances Medium 31633027
2020 Monoubiquitination of FANCD2 (not FANCI) stabilizes the FANCI:FANCD2 heterodimer on dsDNA by clamping; monoubiquitinated FANCI:FANCD2 forms filament-like arrays on long dsDNA as shown by electron microscopy; monoubiquitination does not promote specific exogenous protein-protein interactions. FA pathway reconstitution in vitro, electron microscopy, DNA binding assays, protein-protein interaction screen eLife High 32167469
2020 FANCD2 ubiquitination promotes a large-scale conformational change in the ID2 complex, increasing its affinity for dsDNA by forming a secondary 'Arm' ID2 interface that encircles DNA; FANCI ubiquitination protects the ubiquitin on FANCD2 from USP1-UAF1 deubiquitination. Biochemical ubiquitination assay, DNA binding assays, deubiquitination protection assay, structural analysis EMBO reports High 32510829
2020 ATR directly phosphorylates FANCI on S556, S559, and S565 to stabilize its association with DNA and FANCD2, stimulating ubiquitination of both FANCI and FANCD2 and inhibiting USP1:UAF1 deubiquitination; S559 and S565 are particularly important for protection from deubiquitination. Biochemical reconstitution with purified ATR, FANCI phosphomimetic/phosphodead mutants, in vitro ubiquitination and deubiquitination assays Frontiers in cell and developmental biology High 32117957
2021 Crystal structures of USP1-UAF1 with ubiquitin and cryo-EM of USP1-UAF1 bound to monoubiquitinated FANCI-FANCD2 reveal that USP1-UAF1 drives conformational changes in FANCI-FANCD2 for deubiquitination; UAF1 forms an extensive interface with FANCI required for deubiquitination despite not being directly catalytic. X-ray crystallography (USP1-UAF1), cryo-EM (USP1-UAF1-monoUb-ID complex), mutagenesis, biochemical deubiquitination assay Nature structural & molecular biology High 33795880
2022 ATR phosphorylation of FANCI destabilizes the open state of FANCD2-FANCI and alters its conformational dynamics, priming the clamp for ubiquitination; cryo-EM structures of phosphomimetic FANCI-containing FANCD2-FANCI show complex closure around DNA independent of the FA core complex. Cryo-EM structure determination of phosphomimetic FANCD2-FANCI complexes, DNA binding assays, conformational dynamics analysis Nature structural & molecular biology High 36050501
2024 FANCD2-FANCI acts as a sliding clamp that diffuses on dsDNA and specifically stalls at ss-dsDNA junctions (structures generated at stalled replication forks); cryo-EM structures show that stalled D2-I makes specific contacts with the ss-dsDNA junction distinct from those made when sliding, providing a unified mechanism for recognition and protection of stalled replication forks. Single-molecule imaging (sliding assay), cryo-EM structure determination, in vitro DNA binding at various junction substrates Nature High 39085614
2024 SRSF1 physically interacts with FANCD2 and stimulates its monoubiquitination in an RNA-dependent manner; monoubiquitinated FANCD2 is required for assembly of the SRSF1-NXF1 nuclear export complex and mRNA export; SRSF1 cancer-associated mutants fail to interact with FANCD2, leading to reduced monoubiquitination, decreased mRNA export, and R-loop accumulation. Co-immunoprecipitation, FANCD2 monoubiquitination assay, mRNA export assay, R-loop detection (DRIP), cancer mutant analysis Cell reports Medium 38165804
2014 Fancd2 is required for nuclear retention of Foxo3a in hematopoietic stem cells; Fancd2 deficiency promotes cytoplasmic localization of Foxo3a; re-expression of Fancd2 restores nuclear Foxo3a; this function requires monoubiquitinated Fancd2. Fancd2/Foxo3a double-knockout mouse, immunofluorescence subcellular localization, rescue by Fancd2 re-expression with constitutively active FOXO3a The Journal of biological chemistry Medium 25505262
2018 The N-terminus of USP1 harbors a FANCD2-specific binding sequence required for deubiquitination of K561 on FANCD2; this N-terminal determinant is not required for PCNA or FANCI deubiquitination; the N-terminus alone is sufficient to engineer substrate specificity in a promiscuous USP. In vitro reconstituted deubiquitination of purified monoubiquitinated FANCD2/FANCI/PCNA, USP1 N-terminal truncation/mutants Life science alliance High 30456385
2013 DNA polymerase eta is recruited to DNA damage sites by monoubiquitinated FANCD2 (not the K561R mutant); pol eta interacts with wild-type FANCD2 and this interaction occurs earlier than pol eta-PCNA interaction; FANCD2-null cells expressing FANCD2 or histone H2B-fused pol eta show similar low MMC sensitivity. Co-immunoprecipitation, MMC sensitivity assay, complementation with FANCD2 and pol eta constructs Cell cycle Low 23388460

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2007 Identification of the FANCI protein, a monoubiquitinated FANCD2 paralog required for DNA repair. Cell 583 17412408
2001 Sequence-structure analysis of FAD-containing proteins. Protein science : a publication of the Protein Society 399 11514662
2002 S-phase-specific interaction of the Fanconi anemia protein, FANCD2, with BRCA1 and RAD51. Blood 386 12239151
2002 The FAD- and O(2)-dependent reaction cycle of Ero1-mediated oxidative protein folding in the endoplasmic reticulum. Molecular cell 357 12453408
2004 ATR couples FANCD2 monoubiquitination to the DNA-damage response. Genes & development 330 15314022
2001 Positional cloning of a novel Fanconi anemia gene, FANCD2. Molecular cell 326 11239453
2014 XPF-ERCC1 acts in Unhooking DNA interstrand crosslinks in cooperation with FANCD2 and FANCP/SLX4. Molecular cell 254 24726325
2010 Identification of KIAA1018/FAN1, a DNA repair nuclease recruited to DNA damage by monoubiquitinated FANCD2. Cell 249 20603015
2002 Interaction of FANCD2 and NBS1 in the DNA damage response. Nature cell biology 200 12447395
2004 The DNA crosslink-induced S-phase checkpoint depends on ATR-CHK1 and ATR-NBS1-FANCD2 pathways. The EMBO journal 177 14988723
2007 Deubiquitination of FANCD2 is required for DNA crosslink repair. Molecular cell 165 18082605
2016 FANCD2 Maintains Fork Stability in BRCA1/2-Deficient Tumors and Promotes Alternative End-Joining DNA Repair. Cell reports 163 27264184
2004 Direct interaction of FANCD2 with BRCA2 in DNA damage response pathways. Human molecular genetics 159 15115758
2014 Demystifying the RAD fad. Molecular ecology 140 25319241
2012 The human gene SLC25A17 encodes a peroxisomal transporter of coenzyme A, FAD and NAD+. The Biochemical journal 127 22185573
2011 Structure of the FANCI-FANCD2 complex: insights into the Fanconi anemia DNA repair pathway. Science (New York, N.Y.) 125 21764741
2022 Lactylation: a Passing Fad or the Future of Posttranslational Modification. Inflammation 112 35224683
2003 BRCA1-independent ubiquitination of FANCD2. Molecular cell 112 12887909
2004 Regulated interaction of the Fanconi anemia protein, FANCD2, with chromatin. Blood 110 15454491
2014 The genetic and biochemical basis of FANCD2 monoubiquitination. Molecular cell 101 24905007
2003 Knockdown of zebrafish Fancd2 causes developmental abnormalities via p53-dependent apoptosis. Developmental cell 98 14667412
2010 Hematopoietic stem cell defects in mice with deficiency of Fancd2 or Usp1. Stem cells (Dayton, Ohio) 95 20506303
2015 Measurement of mitochondrial NADH and FAD autofluorescence in live cells. Methods in molecular biology (Clifton, N.J.) 93 25631020
2003 Efficient incorporation of CoA, NAD and FAD into RNA by in vitro transcription. Nucleic acids research 80 12560511
2014 FANCD2 and CtIP cooperate to repair DNA interstrand crosslinks. Cell reports 76 24794434
2017 FAD Regulates CRYPTOCHROME Protein Stability and Circadian Clock in Mice. Cell reports 71 28402850
2011 The E3 ubiquitin ligase RAD18 regulates ubiquitylation and chromatin loading of FANCD2 and FANCI. Blood 70 21355096
2014 CtIP mediates replication fork recovery in a FANCD2-regulated manner. Human molecular genetics 69 24556218
2008 FANCG promotes formation of a newly identified protein complex containing BRCA2, FANCD2 and XRCC3. Oncogene 69 18212739
2012 DNA robustly stimulates FANCD2 monoubiquitylation in the complex with FANCI. Nucleic acids research 68 22287633
2009 A role for monoubiquitinated FANCD2 at telomeres in ALT cells. Nucleic acids research 68 19129235
2018 Replication stress induces accumulation of FANCD2 at central region of large fragile genes. Nucleic acids research 67 29394375
2013 mTOR regulates DNA damage response through NF-κB-mediated FANCD2 pathway in hematopoietic cells. Leukemia 66 23538752
2011 microRNAs: fad or future of liver disease. World journal of gastroenterology 66 21633658
2016 The FANCD2-FANCI complex is recruited to DNA interstrand crosslinks before monoubiquitination of FANCD2. Nature communications 65 27405460
2020 Monoubiquitination by the human Fanconi anemia core complex clamps FANCI:FANCD2 on DNA in filamentous arrays. eLife 63 32167469
2015 FANCI Regulates Recruitment of the FA Core Complex at Sites of DNA Damage Independently of FANCD2. PLoS genetics 62 26430909
2016 BRCA1/FANCD2/BRG1-Driven DNA Repair Stabilizes the Differentiation State of Human Mammary Epithelial Cells. Molecular cell 61 27373334
2018 FANCD2 protects genome stability by recruiting RNA processing enzymes to resolve R-loops during mild replication stress. The FEBS journal 60 30431240
1993 Chromosome 14 and late-onset familial Alzheimer disease (FAD). American journal of human genetics 59 8352272
2019 FAD dependent glucose dehydrogenases - Discovery and engineering of representative glucose sensing enzymes. Bioelectrochemistry (Amsterdam, Netherlands) 56 31838457
2019 DNA requirement in FANCD2 deubiquitination by USP1-UAF1-RAD51AP1 in the Fanconi anemia DNA damage response. Nature communications 54 31253762
2013 FANCD2 activates transcription of TAp63 and suppresses tumorigenesis. Molecular cell 53 23806336
2017 FANCD2 and DNA Damage. International journal of molecular sciences 49 28825622
2018 Arabidopsis thaliana FANCD2 Promotes Meiotic Crossover Formation. The Plant cell 48 29352063
2012 Fanconi anemia proteins FANCD2 and FANCI exhibit different DNA damage responses during S-phase. Nucleic acids research 48 22753026
2013 Recruitment of DNA polymerase eta by FANCD2 in the early response to DNA damage. Cell cycle (Georgetown, Tex.) 47 23388460
2021 Structural basis of FANCD2 deubiquitination by USP1-UAF1. Nature structural & molecular biology 45 33795880
2015 FANCD2 and REV1 cooperate in the protection of nascent DNA strands in response to replication stress. Nucleic acids research 42 26187992
2016 FANCD2 limits BLM-dependent telomere instability in the alternative lengthening of telomeres pathway. Human molecular genetics 41 27427384
2005 Mutation analysis of FANCD2, BRIP1/BACH1, LMO4 and SFN in familial breast cancer. Breast cancer research : BCR 41 16280053
2008 [Internet and cell phone addiction: passing fad or disorder?]. Adicciones 40 18551228
2017 FANCD2 Binds Human Papillomavirus Genomes and Associates with a Distinct Set of DNA Repair Proteins to Regulate Viral Replication. mBio 39 28196964
2019 A Fanci knockout mouse model reveals common and distinct functions for FANCI and FANCD2. Nucleic acids research 38 31219578
2009 The Fanconi anemia protein FANCM is controlled by FANCD2 and the ATR/ATM pathways. The Journal of biological chemistry 38 19633289
2020 Differential functions of FANCI and FANCD2 ubiquitination stabilize ID2 complex on DNA. EMBO reports 37 32510829
2023 FANCD2 inhibits ferroptosis by regulating the JAK2/STAT3 pathway in osteosarcoma. BMC cancer 35 36814203
2020 DXO/Rai1 enzymes remove 5'-end FAD and dephospho-CoA caps on RNAs. Nucleic acids research 35 32374864
2013 The prokaryotic FAD synthetase family: a potential drug target. Current pharmaceutical design 34 23116401
2017 FANCI and FANCD2 have common as well as independent functions during the cellular replication stress response. Nucleic acids research 33 29059323
2007 FANCD2 monoubiquitination provides a link between the HHR6 and FA-BRCA pathways. Cell cycle (Georgetown, Tex.) 31 18277096
2006 Drosophila homologs of FANCD2 and FANCL function in DNA repair. DNA repair 31 16860002
2021 FAD-BERT: Improved prediction of FAD binding sites using pre-training of deep bidirectional transformers. Computers in biology and medicine 30 33601085
2020 ATR-Mediated FANCI Phosphorylation Regulates Both Ubiquitination and Deubiquitination of FANCD2. Frontiers in cell and developmental biology 30 32117957
2007 A requirement of FancL and FancD2 monoubiquitination in DNA repair. Genes to cells : devoted to molecular & cellular mechanisms 29 17352736
2021 Mechanism, specificity, and function of FANCD2-FANCI ubiquitination and deubiquitination. The FEBS journal 28 34137174
2010 FAD binding by ApbE protein from Salmonella enterica: a new class of FAD-binding proteins. Journal of bacteriology 26 21148731
2010 UBE2W interacts with FANCL and regulates the monoubiquitination of Fanconi anemia protein FANCD2. Molecules and cells 26 21229326
2024 The FANCI/FANCD2 complex links DNA damage response to R-loop regulation through SRSF1-mediated mRNA export. Cell reports 25 38165804
2020 FANCD2 Confers a Malignant Phenotype in Esophageal Squamous Cell Carcinoma by Regulating Cell Cycle Progression. Cancers 25 32906798
2017 Overlooked FANCD2 variant encodes a promising, portent tumor suppressor, and alternative polyadenylation contributes to its expression. Oncotarget 25 28157704
2012 Regulation of the Fanconi anemia pathway by a CUE ubiquitin-binding domain in the FANCD2 protein. Blood 25 22855611
2022 The DNA-damage kinase ATR activates the FANCD2-FANCI clamp by priming it for ubiquitination. Nature structural & molecular biology 24 36050501
2020 SLC25A32 sustains cancer cell proliferation by regulating flavin adenine nucleotide (FAD) metabolism. Oncotarget 24 32166001
2019 Clinical importance of FANCD2, BRIP1, BRCA1, BRCA2 and FANCF expression in ovarian carcinomas. Cancer biology & therapy 24 30822218
2018 Specificity for deubiquitination of monoubiquitinated FANCD2 is driven by the N-terminus of USP1. Life science alliance 24 30456385
2017 Involvement of FANCD2 in Energy Metabolism via ATP5α. Scientific reports 24 28687786
2022 Comprehensive analysis of the autophagy-dependent ferroptosis-related gene FANCD2 in lung adenocarcinoma. BMC cancer 23 35236309
2019 Allosteric mechanism for site-specific ubiquitination of FANCD2. Nature chemical biology 23 31873223
2010 FANCJ/BRIP1 recruitment and regulation of FANCD2 in DNA damage responses. Chromosoma 23 20676667
2006 An NADPH and FAD dependent enzyme catalyzes hydroxylation of flavonoids in position 8. Phytochemistry 23 16678869
2019 The hidden side of the human FAD synthase 2. International journal of biological macromolecules 22 31351152
2019 Nuclear receptors regulate alternative lengthening of telomeres through a novel noncanonical FANCD2 pathway. Science advances 22 31633027
2017 Fanconi anemia FANCD2 and FANCI proteins regulate the nuclear dynamics of splicing factors. The Journal of cell biology 22 29030393
2024 FANCD2-FANCI surveys DNA and recognizes double- to single-stranded junctions. Nature 21 39085614
2021 Natural diversity of FAD-dependent 4-hydroxybenzoate hydroxylases. Archives of biochemistry and biophysics 21 33684360
2023 LncRNA SNHG1 upregulates FANCD2 and G6PD to suppress ferroptosis by sponging miR-199a-5p/3p in hepatocellular carcinoma. Drug discoveries & therapeutics 20 37599085
2016 Chromosomal Integrity after UV Irradiation Requires FANCD2-Mediated Repair of Double Strand Breaks. PLoS genetics 20 26765540
2015 FANCD2 influences replication fork processes and genome stability in response to clustered DSBs. Cell cycle (Georgetown, Tex.) 20 26083937
2012 Heterozygote FANCD2 mutations associated with childhood T Cell ALL and testicular seminoma. Familial cancer 20 22829014
2006 Developmental stage- and DNA damage-specific functions of C. elegans FANCD2. Biochemical and biophysical research communications 20 17126808
2014 The Fanconi anemia proteins FANCD2 and FANCJ interact and regulate each other's chromatin localization. The Journal of biological chemistry 19 25070891
2009 FANCD2 hurdles the DNA interstrand crosslink. Cell 19 20064367
2021 FANCD2 limits acetaldehyde-induced genomic instability during DNA replication in esophageal keratinocytes. Molecular oncology 17 34328261
2021 Utilization of FAD-Glucose Dehydrogenase from T. emersonii for Amperometric Biosensing and Biofuel Cell Devices. Analytical chemistry 17 34383460
2021 FAN1's protection against CGG repeat expansion requires its nuclease activity and is FANCD2-independent. Nucleic acids research 17 34718701
2017 The identification of FANCD2 DNA binding domains reveals nuclear localization sequences. Nucleic acids research 17 28666371
2014 Fancd2 is required for nuclear retention of Foxo3a in hematopoietic stem cell maintenance. The Journal of biological chemistry 16 25505262
2020 Structural insight into FANCI-FANCD2 monoubiquitination. Essays in biochemistry 15 32725171
2017 Beyond interstrand crosslinks repair: contribution of FANCD2 and other Fanconi Anemia proteins to the replication of DNA. Mutation research 15 29031493