{"gene":"WDHD1","run_date":"2026-06-11T09:02:06","timeline":{"discoveries":[{"year":1997,"finding":"AND-1 (WDHD1) was identified and characterized as a novel nuclear DNA-binding protein containing an N-terminal WD-repeat domain and a C-terminal HMG-box domain. It binds four-way junction DNA, exists as an oligomer (likely homodimer), and localizes to the interchromatinic space during interphase but disperses to the cytoplasm during mitosis.","method":"cDNA cloning, monoclonal antibody characterization, DNA affinity chromatography, EMSA, immunolocalization","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal biochemical methods in a single study establishing domain architecture, DNA binding, and localization","pmids":["9175701"],"is_preprint":false},{"year":1992,"finding":"CTF4/CHL15 (yeast ortholog of WDHD1) encodes a ~105 kDa protein that associates in vitro with DNA polymerase alpha; ctf4 mutants show elevated sister chromatid loss and recombination but not elevated point mutation rates, and the C-terminal helix-loop-helix domain is essential for CTF4 function.","method":"Yeast genetics, chromosome segregation assays, in vitro DNA polymerase alpha association, sequence analysis, frameshift mutagenesis","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic and biochemical methods in a single lab establishing Pol alpha association and functional domain requirements","pmids":["1341195"],"is_preprint":false},{"year":1999,"finding":"Yeast CTF4 genetically interacts with DNA2 and POL1 (Pol alpha subunit); ctf4 deletion is synthetically lethal with DNA damage-sensitive dna2 alleles, placing Ctf4 in a pathway with Dna2 and Pol alpha for lagging strand synthesis or repair.","method":"Yeast genetic epistasis, synthetic lethality analysis, allele construction","journal":"Genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with multiple alleles establishing pathway position","pmids":["10101169"],"is_preprint":false},{"year":2001,"finding":"Yeast CTF4 is required for sister chromatid cohesion; ctf4 null mutants exhibit cohesion failure and preanaphase accumulation dependent on the spindle assembly checkpoint, placing CTF4 functionally at the replication fork in the cohesion establishment pathway.","method":"Yeast genetics, sister chromatid cohesion assay, checkpoint epistasis","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — replicated across labs, cohesion assay with checkpoint epistasis establishing pathway position","pmids":["11287619"],"is_preprint":false},{"year":2002,"finding":"Fission yeast Mcl1 (ortholog of CTF4/WDHD1) is required for chromosome replication, sister chromatid cohesion, and segregation; mcl1-1 is synthetically lethal with checkpoint mutations rad3Δ and rad26Δ, and interacts genetically with replication components.","method":"Fission yeast genetics, pulsed-field electrophoresis, synthetic lethality, overexpression analysis","journal":"Eukaryotic cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis and cell biological phenotypes in a single lab","pmids":["12455694"],"is_preprint":false},{"year":2004,"finding":"Yeast CTF4 (polymerase-alpha-associated protein) is required for efficient sister chromatid cohesion in both mitosis and meiosis; ctf4Δ cells show massive meiosis II non-disjunction and loss of centromeric cohesion despite normal cohesin association with centromeric DNA.","method":"Yeast genetics, cohesion assays, cytological analysis of meiosis","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple phenotypic assays in a single lab establishing meiotic cohesion requirement","pmids":["15226378"],"is_preprint":false},{"year":2007,"finding":"Human AND-1/CTF4 (WDHD1) interacts with Mcm10 (which associates with MCM2-7) and with the p180 subunit of DNA polymerase alpha. In Xenopus egg extracts, AND-1 loads on chromatin after Mcm10, concurrently with Pol alpha, and is required for efficient DNA synthesis. Mcm10 is required for chromatin loading of AND-1 and Pol alpha; disrupting the Mcm10-AND-1 interaction inhibits DNA synthesis.","method":"Co-immunoprecipitation, Xenopus egg extract DNA replication assay, chromatin fractionation, inhibitory antibody, siRNA knockdown in mammalian cells","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (Co-IP, antibody inhibition, egg extract reconstitution, siRNA) across human and Xenopus systems","pmids":["17761813"],"is_preprint":false},{"year":2009,"finding":"Yeast Ctf4 couples the MCM2-7 helicase to DNA polymerase alpha within the replisome progression complex (RPC); Ctf4 and GINS together are crucial for this coupling. In ctf4Δ cells, Pol alpha is unstable and barely detectable at replication forks, while helicase (and Pol epsilon) quantities are unchanged but their localizations are abnormally dispersed. Cells lacking both Ctf4 and Mrc1 experience chronic DNA damage checkpoint activation and fail to complete the cell cycle.","method":"Yeast genetics, ChIP, in vitro interaction assays between GINS and Pol alpha mediated by Ctf4, immunoprecipitation, double mutant analysis","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — ChIP, in vitro reconstitution of GINS-Ctf4-Pol alpha interaction, and genetic double-mutant analysis in a single study","pmids":["19661920"],"is_preprint":false},{"year":2009,"finding":"Yeast Ctf4 mediates the in vitro interaction between GINS and DNA Pol alpha-primase; in ctf4Δ mutants, the MCM-GINS–Pol alpha interaction is specifically destabilized, Pol alpha becomes unstable at replication forks, and the replication checkpoint is activated. Tof1 and Mrc1 are not required for this interaction.","method":"Yeast genetics, in vitro protein interaction assay, ChIP, immunoblot","journal":"Genes to cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro interaction plus ChIP in a single lab","pmids":["19496828"],"is_preprint":false},{"year":2009,"finding":"Human AND-1/CTF4 (WDHD1) is required for assembly of the CMG (Cdc45-Mcm2-7-GINS) complex in human cells; along with RecQL4 and Mcm10, AND-1 is needed for stable CMG interactions observed only after G1/S transition and dependent on CDK and Cdc7 kinase activities.","method":"Bimolecular fluorescence complementation (BiFC) in HeLa cells, siRNA depletion, CDK and Cdc7 kinase inhibition","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — BiFC assay with siRNA and kinase inhibitor controls in a single lab","pmids":["19805216"],"is_preprint":false},{"year":2009,"finding":"Human AND-1 (WDHD1) localizes adjacent to replication foci in S phase, is phosphorylated in response to replication arrest in a checkpoint kinase (ATM, ATR, Cdc7)-dependent manner, interacts with cohesin proteins Smc1, Smc3, and Rad21/Scc1, and is required for homologous recombination repair. Depletion of AND-1 increases DNA damage, delays S-phase progression, compromises checkpoint activation, and decreases Chk1 protein levels.","method":"Immunofluorescence, co-immunoprecipitation, siRNA knockdown, HR repair assay (I-SceI DSB), immunoblot, flow cytometry","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods in a single lab establishing localization and interaction with cohesin","pmids":["19439411"],"is_preprint":false},{"year":2010,"finding":"Purified human CTF4 (WDHD1) exists as a dimer; the SepB domain determines dimeric structure. hCtf4 binds preferentially to DNA template-primer structures, directly interacts with DNA polymerases alpha, delta, and epsilon, and markedly stimulates Pol alpha and Pol epsilon activities in vitro. siRNA depletion of hCtf4 in HeLa cells causes G1/S arrest and slower DNA replication.","method":"Protein purification, in vitro DNA polymerase stimulation assay, EMSA/DNA binding, co-immunoprecipitation, siRNA knockdown, DNA fiber analysis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro biochemical reconstitution of Pol stimulation with purified proteins, plus multiple orthogonal methods","pmids":["20089864"],"is_preprint":false},{"year":2010,"finding":"Xenopus Dna2 forms a complex with And-1/Ctf4 and Mcm10 at replication forks (demonstrated by co-immunoprecipitation from egg extracts), and is recruited to DNA after replication origin licensing.","method":"Xenopus egg extract, co-immunoprecipitation, immunofluorescence/co-localization","journal":"Cell cycle","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-IP in egg extracts, replicated in context of a broader study","pmids":["20237432"],"is_preprint":false},{"year":2011,"finding":"Human WDHD1 associates with centromeres in a cell cycle-dependent manner (mid-to-late S phase), and its down-regulation compromises HP1alpha localization to pericentric heterochromatin, alters epigenetic marks at pericentric chromatin, disrupts mitosis, and impairs generation of centromeric satellite small non-coding RNAs by reducing Dicer association with centromeric RNA.","method":"ChIP, immunofluorescence, siRNA knockdown, chromatin fractionation, RNA analysis","journal":"Nucleic acids research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (ChIP, IF, RNA analysis) in a single lab","pmids":["21266480"],"is_preprint":false},{"year":2011,"finding":"AND-1 (WDHD1) forms a complex with both histone H3 and histone acetyltransferase Gcn5; AND-1 down-regulation causes Gcn5 protein degradation, reducing H3K9 and H3K56 acetylation. AND-1 overexpression stabilizes Gcn5 through protein-protein interaction.","method":"Co-immunoprecipitation, siRNA knockdown, immunoblot, overexpression","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP and knockdown/overexpression in a single lab; no reconstitution with purified proteins","pmids":["21725360"],"is_preprint":false},{"year":2012,"finding":"Mimosine prevents chromatin binding of Ctf4/AND-1 in HeLa cells, arresting the cell cycle in G1 before S-phase entry via a Hif-1alpha-dependent increase in p27; depletion of Hif-1alpha restores Ctf4 chromatin binding and allows S-phase entry even in the presence of mimosine.","method":"Chromatin fractionation, siRNA knockdown, cell cycle analysis, flow cytometry","journal":"Cell cycle","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — chromatin binding assay plus siRNA rescue establishing pathway","pmids":["22374673"],"is_preprint":false},{"year":2013,"finding":"Human CTF4 (WDHD1) interacts with the CMG (Cdc45-Mcm2-7-GINS) complex; the hCtf4-CMG complex was isolated by in vitro interaction of purified proteins, by co-infection of insect cells, and from HeLa cell chromatin. The complex contains homodimeric hCtf4 and monomeric CMG. The hCtf4-CMG complex retains DNA helicase activity that is more salt-resistant than CMG alone. The stability of hCtf4-CMG depends on interactions between hCtf4 and multiple CMG components.","method":"Purified protein interaction, Sf9 co-infection/co-purification, HeLa chromatin immunoprecipitation, helicase assay","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Strong — reconstituted with purified proteins, validated in insect cells and from human chromatin, enzymatic assay","pmids":["24255107"],"is_preprint":false},{"year":2014,"finding":"Yeast Ctf4 self-associates as a constitutive disk-shaped homotrimer via a C-terminal beta-propeller (SepB) domain fused to a helical extension. Pol alpha catalytic subunit and GINS subunit Sld5 share a conserved Ctf4-interacting peptide (CIP-box) that docks onto the helical extension of a Ctf4 protomer; one Ctf4 trimer can simultaneously bind both Pol alpha and GINS, enabling coupling of two Pol alpha molecules to one CMG helicase.","method":"X-ray crystallography, electron microscopy, native mass spectrometry, mutagenesis, in vitro binding assays","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure and EM with functional mutagenesis and native MS, multiple orthogonal methods in a rigorous single study","pmids":["24805245"],"is_preprint":false},{"year":2015,"finding":"Yeast Pol alpha/Primase/Ctf4 mutants that are proficient in bulk DNA replication are defective in recombination-mediated damage bypass by template switching and have reduced sister chromatid cohesion; these DDT defects are caused by altered ssDNA metabolism and abnormal replication fork topology rather than increased sister chromatid distance.","method":"Yeast genetics, DNA fiber analysis, electron microscopy of replication intermediates, sister chromatid cohesion assay","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (genetics, EM of fork structures, fiber analysis) establishing mechanistic distinction","pmids":["25661486"],"is_preprint":false},{"year":2015,"finding":"RecQL4 is required for the association of Mcm10 and Ctf4 with replication origins in human cells; physical interactions between RecQL4, Mcm10, and Ctf4 and their origin association require both CDK and DDK activities and are blocked by the DNA damage checkpoint.","method":"ChIP at replication origins, co-immunoprecipitation, siRNA knockdown, kinase inhibition","journal":"Cell cycle","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP and Co-IP with kinase inhibitor controls in a single lab","pmids":["25602958"],"is_preprint":false},{"year":2015,"finding":"In response to replication stress, AND-1 is phosphorylated at T826 by ATR; this phosphorylation causes AND-1 to accumulate at damage sites where it promotes the Claspin-Chk1 interaction, stimulating efficient Chk1 activation by ATR. AND-1 directly binds ssDNA and facilitates Claspin association with ssDNA. AND-1 is required for recovery of stalled forks.","method":"Phosphorylation site mutagenesis, co-immunoprecipitation, in vitro ssDNA binding assay, siRNA knockdown, immunofluorescence, DNA fiber analysis","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — phospho-site mutagenesis combined with Co-IP, in vitro ssDNA binding, and functional rescue in a single study","pmids":["26082189"],"is_preprint":false},{"year":2015,"finding":"Histone H3K56 acetylation acts through Ctf4 to uncouple CMG helicase from DNA polymerases under replication stress; the N-terminal domain of Ctf4 interacts with Mms22 (adaptor of the Rtt101-Mms1 E3 ubiquitin ligase), and this interaction is promoted by replicative stress.","method":"Yeast genetics, genetic epistasis, domain deletion analysis, two-hybrid/co-immunoprecipitation","journal":"Genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis and domain analysis in a single lab","pmids":["25697176"],"is_preprint":false},{"year":2016,"finding":"Ctf4 recruits the Chl1 helicase to the replisome via a conserved CIP-box interaction motif shared with GINS and Pol alpha; EM analysis visualized a reconstituted Chl1-Ctf4-GINS assembly. Ctf4 interaction (but not Chl1 helicase activity) is required for sister chromatid cohesion, while Chl1 helicase activity is required for replication fork progression under nucleotide depletion. Physical interaction between Chl1 and cohesin occurs during S phase.","method":"Electron microscopy of reconstituted complex, in vitro binding assays, yeast genetics with separation-of-function mutants, co-immunoprecipitation","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1 / Strong — EM-validated reconstituted complex, in vitro binding, separation-of-function genetics, and Co-IP across multiple methods","pmids":["27397686"],"is_preprint":false},{"year":2016,"finding":"Ctf4 acts as a hub connecting multiple CIP-box proteins (in addition to Pol alpha and GINS) to the CMG helicase; crystallographic analysis classifies CIP-boxes into two groups targeting different sites on Ctf4. Ctf4-dependent recruitment of Dna2 and Tof2 via their CIP-boxes is required for rDNA copy-number regulation but not genome-wide DNA synthesis.","method":"X-ray crystallography, in vitro binding assays, yeast genetics, CIP-box mutagenesis, genome-wide replication analysis","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structures with functional mutagenesis and genetics establishing multi-partner hub mechanism","pmids":["27397685"],"is_preprint":false},{"year":2016,"finding":"WDHD1 (AND-1) plays an important role in E7 (HPV-16)-induced G1 checkpoint abrogation and rereplication; WDHD1 protein steady-state level and half-life are increased in E7-expressing cells, and WDHD1 knockdown reduces E7-induced G1 checkpoint abrogation.","method":"siRNA knockdown, immunoblot (protein stability), cell cycle analysis, rereplication assay","journal":"Journal of virology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with specific cell cycle phenotype in a single lab","pmids":["27099318"],"is_preprint":false},{"year":2017,"finding":"Human AND-1 (WDHD1) maintains the trimeric structure of yeast Ctf4 via its conserved SepB domain, but interacts with DNA Pol alpha/primase primarily through its C-terminal HMG box, which binds the Pol alpha B subunit at the same site targeted by SV40 T-antigen. AND-1 also possesses a DNA-binding activity via its HMG box that may position Pol alpha on the lagging-strand template.","method":"Biochemical protein interaction assays, domain mutagenesis, competition with SV40 T-antigen peptide, DNA binding assay","journal":"Open biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro biochemical reconstitution with mutagenesis and competition experiments establishing interaction mechanism","pmids":["29167311"],"is_preprint":false},{"year":2017,"finding":"Human AND-1 (WDHD1) exists as a homotrimer mediated by the SepB domain; the positively charged groove within SepB provides binding sites for Pol alpha. AND-1 binds DNA in vitro via its C-terminal HMG domain. Unlike yeast Ctf4, human AND-1's recruitment to the CMG complex appears to be mediated by unknown participants other than GINS.","method":"X-ray crystallography of WD40 and SepB domains, biochemical assays, mutagenesis, DNA binding assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure with biochemical validation and mutagenesis in a single study","pmids":["28381552"],"is_preprint":false},{"year":2017,"finding":"AND-1/Ctf4 promotes DNA end resection and homologous recombination repair by interacting with CtIP and regulating its recruitment to DNA damage sites. AND-1 localizes to damage sites via the MDC1-RNF8 pathway and is required for sustained ATR-CHK1 signaling and maintenance of intra-S and G2-phase checkpoints.","method":"Co-immunoprecipitation, siRNA knockdown, immunofluorescence at damage foci, HR repair assay, checkpoint assays","journal":"Nucleic acids research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple methods (Co-IP, IF, functional assays) in a single lab","pmids":["27940552"],"is_preprint":false},{"year":2017,"finding":"AND-1 (WDHD1) forms complexes with CtIP and other repair proteins; AND-1 is recruited to DSB sites dependent on MDC1, BRCA1, and ATM. AND-1 knockdown impairs DSB end resection by reducing CtIP recruitment, reduces Chk1 activation, establishing an MDC1→AND-1→CtIP axis for CtIP-mediated end resection.","method":"Co-immunoprecipitation, siRNA knockdown, immunofluorescence at damage foci, end resection assay, Chk1 activation assay","journal":"Nucleic acids research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple methods in a single lab; replicates the Chen et al. 2017 finding from a different group","pmids":["27940557"],"is_preprint":false},{"year":2017,"finding":"Yeast Ctf4 prevents genome rearrangements by suppressing DSB formation and end resection at arrested replication forks; in ctf4Δ cells, DSBs form more frequently at arrested forks, undergo end resection, and are repaired by HR prone to rDNA hyper-amplification.","method":"Yeast genetics, Southern blotting/rDNA copy number, DSB quantification, end resection assay","journal":"Molecular cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic and molecular analysis of DSB end resection with multiple assays in a single lab","pmids":["28525744"],"is_preprint":false},{"year":2017,"finding":"A stapled peptide encoding the Ctf4-interacting peptide (CIP) of Sld5 binds Ctf4 with submicromolar affinity; crystal structure confirmed the mode of interaction; the stapled peptide displaces Pol alpha from the replisome in yeast extracts.","method":"Stapled peptide design, crystal structure, fluorescence polarization binding assay, yeast extract replisome disruption assay","journal":"Angewandte Chemie","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure plus functional displacement assay with purified/extract proteins","pmids":["28815832"],"is_preprint":false},{"year":2017,"finding":"ATR-Chk1 inhibitor-induced unscheduled origin firing in human cells is mediated by Cdc7 kinase through phosphorylations on GINS that induce an association between GINS and AND-1 (WDHD1).","method":"Proteomics (phosphoproteomics), co-immunoprecipitation, kinase inhibition, siRNA knockdown","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — phosphoproteomic identification with Co-IP validation and kinase inhibitor evidence in a single lab","pmids":["29123096"],"is_preprint":false},{"year":2018,"finding":"The Mcm2-Ctf4-Pol alpha axis facilitates transfer of parental histone (H3-H4)2 tetramers to lagging-strand DNA at replication forks; mutations disrupting the Mcm2 histone-binding domain, or Ctf4-Pol alpha connection, result in enrichment of parental histones on leading strand DNA.","method":"SCAR-seq (strand-specific parental histone enrichment), yeast genetics with separation-of-function mutations, ChIP","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — strand-specific histone tracking with multiple separation-of-function mutants across Mcm2, Ctf4, and Pol alpha in a single rigorous study","pmids":["30244834"],"is_preprint":false},{"year":2018,"finding":"AND-1 (WDHD1) protects replication forks from resection; depletion of AND-1 using an inducible degron causes fork speed slow-down, accumulation of long ssDNA gaps at the fork junction, and conversion of these to DSBs in G2 via MRE11 nuclease activity. The WD40 domain of AND-1 is specifically required for fork protection and proliferation, while the HMG box is required for fast replication but not proliferation.","method":"Auxin-inducible degron (AND-1 depletion), DNA fiber analysis, electron microscopy of replication intermediates, MRE11 inhibitor (mirin), domain-specific mutants","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — inducible degron system with domain separation-of-function, EM of fork structures, and pharmacological validation in a single rigorous study","pmids":["30082684"],"is_preprint":false},{"year":2019,"finding":"Ctf4 tightly dimerizes CMG helicase via an interface involving Psf2, Cdc45, and Sld5; Ctf4 binds only one Pol alpha-primase per trimer, suggesting a replication factory model where two CMG helicases and one Pol alpha-primase are organized by one Ctf4 trimer. Cryo-EM reveals two CMGs nearly facing each other with lagging strands toward the center.","method":"Cryo-EM structure, in vitro reconstitution, pulldown assays","journal":"eLife","confidence":"High","confidence_rationale":"Tier 1 / Strong — cryo-EM structure with in vitro reconstitution establishing stoichiometry and architecture","pmids":["31589141"],"is_preprint":false},{"year":2019,"finding":"Human CST (CTC1-STN1-TEN1) promotes AND-1/Ctf4 chromatin association and replisome assembly; CST directly interacts with AND-1 and pol alpha, enhancing their chromatin association independently of exogenous replication stress.","method":"Co-immunoprecipitation, chromatin fractionation, siRNA knockdown, origin licensing assays","journal":"Life science alliance","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP with chromatin fractionation in a single lab","pmids":["30979824"],"is_preprint":false},{"year":2020,"finding":"CryoEM structure of human CMG bound to AND-1 (CMGA) reveals the AND-1 trimer docks onto the helicase assembly formed by Cdc45 and GINS via one beta-propeller domain of its trimerization region; in the CMGA architecture, AND-1 is closely positioned to fork DNA while its CIP-binding helical domains remain available for partner protein recruitment.","method":"CryoEM structure at 3.3 Å resolution","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 1 / Strong — high-resolution cryoEM structure establishing the molecular docking interface","pmids":["32453425"],"is_preprint":false},{"year":2020,"finding":"Yeast Sen1 (transcription termination helicase) associates with the replisome via Ctf4 and Mrc1; the N-terminus of Sen1 binds Ctf4 and Mrc1. A separation-of-function mutant (sen1-3) that abolishes replisome binding without affecting transcription termination shows increased genome instability and recombination.","method":"Yeast two-hybrid, co-immunoprecipitation, separation-of-function mutagenesis, genetics","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — separation-of-function mutant with Co-IP validation in a single lab","pmids":["32075754"],"is_preprint":false},{"year":2020,"finding":"Ctf4-mediated Pol alpha recruitment for lagging-strand initiation and for origin firing are both sensitive to Pol alpha abundance, but lagging-strand initiation is specifically more dependent on Ctf4-Pol alpha interaction when Pol alpha is limiting, suggesting Pol alpha associates distributively at the fork.","method":"Yeast genetics with titratable Pol alpha levels, Okazaki fragment analysis, origin firing efficiency measurement, Ctf4-Pol alpha interaction mutants","journal":"PLoS genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo genetic titration with Okazaki fragment sequencing in a single study","pmids":["32379761"],"is_preprint":false},{"year":2020,"finding":"WDHD1 (AND-1) acts as an E3 ubiquitin ligase that promotes ubiquitination of MAPRE2 in the nucleus; WDHD1 and MAPRE2 interact by co-immunoprecipitation, and WDHD1 overexpression leads to MAPRE2 degradation conferring cisplatin resistance in lung adenocarcinoma cells.","method":"Co-immunoprecipitation, ubiquitination assay, WDHD1 knockout, protein stability assay, xenograft","journal":"Frontiers in oncology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP and ubiquitination assay in a single lab; E3 ligase activity not biochemically reconstituted with purified proteins","pmids":["32426268"],"is_preprint":false},{"year":2021,"finding":"STAT3 transcription factor binds the promoter/upstream regulatory region of WDHD1 gene and activates its transcription; WDHD1 mediates the DNA replication function of STAT3 (STAT3 knockdown reduces DNA replication, which is rescued by WDHD1 expression).","method":"ChIP (STAT3 at WDHD1 promoter), siRNA knockdown, ectopic expression, DNA replication assay","journal":"Cell & bioscience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP plus epistasis rescue experiment in a single lab","pmids":["33413624"],"is_preprint":false},{"year":2021,"finding":"Two AND-1 inhibitors (bazedoxifene acetate and compound CH3) promote AND-1 degradation by disrupting AND-1 polymerization through direct interaction with the WD40 domain; depolymerization promotes interaction with E3 ligase CUL4B, leading to AND-1 ubiquitination and degradation.","method":"High-throughput screen, in vitro binding assay (WD40 domain interaction), co-immunoprecipitation (CUL4B interaction), ubiquitination assay, in vitro and in vivo cancer growth assays","journal":"Clinical and translational medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct binding to WD40 domain and Co-IP for CUL4B interaction established, ubiquitination shown; reconstitution not fully purified","pmids":["34923765"],"is_preprint":false},{"year":2022,"finding":"AND-1 (WDHD1) is phosphorylated at T826 by ATR in response to interstrand crosslinks (ICLs); this phosphorylation triggers an intramolecular change that promotes AND-1 interaction with FANCM/FAAP24 complex, recruiting it to ICL-stalled forks to activate Fanconi anemia signaling.","method":"Co-immunoprecipitation, phosphorylation site mutagenesis (T826A), ATR inhibition, immunofluorescence at ICL foci, proximity ligation assay","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Strong — phospho-site mutagenesis with Co-IP, PLA, and functional FA pathway assays; extends the ATR-phosphorylation mechanism established in the 2015 EMBO J paper","pmids":["35867033"],"is_preprint":false},{"year":2024,"finding":"ZBTB16 transcription factor inhibits WDHD1 transcription; ectopic WDHD1 expression reverses ZBTB16-mediated inhibition of lung adenocarcinoma cell proliferation and S-phase cell cycle arrest, placing WDHD1 downstream of ZBTB16 in a regulatory axis.","method":"Ectopic expression, siRNA/overexpression rescue, cell cycle analysis, xenograft, bioinformatics/promoter analysis","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis (WDHD1 rescue of ZBTB16 overexpression phenotype) in a single lab","pmids":["38654107"],"is_preprint":false}],"current_model":"WDHD1/AND-1/CTF4 is a homotrimeric replisome hub protein (trimerization driven by its SepB/beta-propeller domain) that bridges the CMG (Cdc45-MCM-GINS) helicase to DNA polymerase alpha-primase via CIP-box interactions on its helical extensions, thereby coupling DNA unwinding to lagging-strand priming; it is loaded onto chromatin in a Mcm10- and RecQL4-dependent manner after the G1/S transition, stimulates the activities of replicative polymerases in vitro, facilitates parental histone H3-H4 transfer to lagging strands, recruits multiple CIP-box partners (including Chl1 helicase, Dna2, and Sen1) to coordinate sister chromatid cohesion establishment, rDNA copy-number maintenance, and transcription-replication conflict resolution; in response to replication stress, AND-1 is phosphorylated by ATR at T826, enabling it to accumulate at stalled forks and promote Claspin-Chk1 checkpoint activation and FANCM/FAAP24-dependent Fanconi anemia signaling; its WD40 domain specifically protects replication forks from MRE11-dependent resection and DSB formation, while its HMG box confers DNA-binding activity and mediates the unique mammalian interaction with the Pol alpha B subunit."},"narrative":{"mechanistic_narrative":"WDHD1 (AND-1/CTF4) is a replisome hub protein that physically couples the CMG (Cdc45-MCM2-7-GINS) helicase to DNA polymerase alpha-primase, thereby linking parental-strand unwinding to lagging-strand priming and coordinating downstream genome-maintenance events [PMID:19661920, PMID:24255107, PMID:24805245]. It self-associates into a disk-shaped homotrimer through a C-terminal beta-propeller/SepB domain, and the helical extensions of each protomer dock conserved Ctf4-interacting peptides (CIP-boxes) shared by Pol alpha and the GINS subunit Sld5, allowing one trimer to bridge multiple replisome components [PMID:24805245, PMID:28381552]; cryo-EM of the human CMG-AND-1 (CMGA) assembly shows the trimer docking onto Cdc45/GINS via one propeller while its CIP-binding helices remain free for partner recruitment [PMID:32453425], and the trimer can dimerize two CMG helicases around a central Pol alpha-primase to form a replication factory [PMID:31589141]. In human cells AND-1 also engages Pol alpha through its HMG box, which binds the Pol alpha B subunit at the SV40 T-antigen site and confers DNA-binding activity that positions Pol alpha on the lagging-strand template [PMID:29167311]. Loaded onto chromatin after the G1/S transition in an Mcm10- and RecQL4-dependent, CDK/DDK-controlled manner, AND-1 is required for stable CMG assembly and stimulates replicative polymerase activity in vitro [PMID:17761813, PMID:19805216, PMID:25602958, PMID:20089864]. As a CIP-box hub it recruits the Chl1 helicase, Dna2, and the transcription-termination helicase Sen1 to the fork to support sister chromatid cohesion establishment, rDNA copy-number control, and transcription-replication conflict resolution [PMID:27397686, PMID:27397685, PMID:32075754, PMID:11287619], and through the Mcm2-Ctf4-Pol alpha axis it directs transfer of parental H3-H4 to lagging strands [PMID:30244834]. Under replication stress, ATR phosphorylates AND-1 at T826, driving its accumulation at stalled forks to promote Claspin-Chk1 checkpoint activation and, at interstrand crosslinks, FANCM/FAAP24-dependent Fanconi anemia signaling [PMID:26082189, PMID:35867033]; its WD40 domain specifically protects forks from MRE11-dependent resection and DSB formation [PMID:30082684].","teleology":[{"year":1992,"claim":"Established the founding link between this protein family and the replication machinery by showing the yeast ortholog associates with Pol alpha and is needed for faithful chromosome transmission.","evidence":"Yeast genetics and in vitro Pol alpha association with CTF4/CHL15","pmids":["1341195"],"confidence":"Medium","gaps":["No structural basis for the Pol alpha interaction","Mechanism connecting Pol alpha binding to chromosome loss unresolved"]},{"year":1997,"claim":"Defined the human protein's domain architecture and biochemical properties, identifying an N-terminal WD-repeat domain, a C-terminal HMG box, four-way-junction DNA binding, and cell-cycle-regulated nuclear localization.","evidence":"cDNA cloning, DNA affinity chromatography, EMSA, and immunolocalization","pmids":["9175701"],"confidence":"Medium","gaps":["Replisome role not yet established","Oligomeric state mischaracterized as dimer","No functional assignment to domains"]},{"year":2004,"claim":"Genetically positioned the protein in the sister chromatid cohesion establishment pathway acting at the replication fork in both mitosis and meiosis.","evidence":"Yeast cohesion and checkpoint-epistasis assays across mitosis and meiosis","pmids":["11287619","12455694","15226378","10101169"],"confidence":"High","gaps":["Molecular mechanism coupling fork passage to cohesion unknown","Did not identify the direct effectors recruited for cohesion"]},{"year":2009,"claim":"Demonstrated the central coupling function: the protein bridges the MCM/GINS helicase to Pol alpha within the replisome, and is required for CMG assembly and replisome stability in human cells.","evidence":"Yeast ChIP and in vitro GINS-Ctf4-Pol alpha reconstitution; human BiFC with siRNA and kinase inhibition; Xenopus extract loading after Mcm10","pmids":["19661920","19496828","19805216","17761813"],"confidence":"High","gaps":["Atomic interaction interfaces not yet defined","Stoichiometry of the coupling not determined"]},{"year":2010,"claim":"Reconstituted the biochemical activity, showing the purified protein binds template-primer DNA, directly contacts Pol alpha/delta/epsilon, and stimulates Pol alpha and epsilon activity.","evidence":"Purified-protein DNA-binding and polymerase-stimulation assays plus DNA fiber analysis","pmids":["20089864","20237432"],"confidence":"High","gaps":["Oligomeric state reported as dimer, later revised to trimer","How polymerase stimulation operates at the fork in vivo unclear"]},{"year":2014,"claim":"Solved the architecture as a homotrimeric hub: a SepB beta-propeller drives trimerization and helical extensions dock shared CIP-box peptides from Pol alpha and GINS, enabling simultaneous binding of multiple replisome partners.","evidence":"X-ray crystallography, EM, native mass spectrometry, and mutagenesis in yeast; human SepB/WD40 crystal structures","pmids":["24805245","24255107","28381552","26082189"],"confidence":"High","gaps":["In vivo consequences of trimer disruption not fully mapped","Human CMG recruitment route differed from yeast and remained undefined"]},{"year":2016,"claim":"Generalized the hub model by showing the trimer recruits additional CIP-box partners (Chl1, Dna2, Tof2, Sen1) to assign distinct genome-maintenance outputs including cohesion, rDNA copy-number control, and conflict resolution.","evidence":"Crystallography, EM of reconstituted assemblies, and separation-of-function yeast genetics","pmids":["27397686","27397685","32075754"],"confidence":"High","gaps":["Competition/hierarchy among CIP-box partners at a single fork unresolved","Human counterparts of several yeast partner interactions not tested"]},{"year":2017,"claim":"Defined the mammalian-specific Pol alpha contact through the HMG box binding the Pol alpha B subunit at the SV40 T-antigen site, distinguishing human AND-1 mechanism from yeast.","evidence":"Biochemical interaction assays, domain mutagenesis, and T-antigen peptide competition","pmids":["29167311"],"confidence":"High","gaps":["How HMG-box and SepB contacts are coordinated on one trimer unclear","Physiological requirement for the B-subunit contact in cells untested here"]},{"year":2017,"claim":"Connected the protein to the DNA-damage response, showing ATR phosphorylation at T826 drives accumulation at stalled forks to promote Claspin-Chk1 activation and that it regulates CtIP-mediated end resection and HR repair.","evidence":"Phospho-site mutagenesis, in vitro ssDNA binding, Co-IP, immunofluorescence at damage foci, and HR/resection assays","pmids":["26082189","27940552","27940557","28525744"],"confidence":"High","gaps":["How a replisome coupler is repurposed for resection mechanistically incomplete","Relationship between fork-protective and resection-promoting roles unresolved"]},{"year":2018,"claim":"Assigned domain-specific genome-protection functions: the Mcm2-Ctf4-Pol alpha axis directs parental H3-H4 transfer to lagging strands, and the WD40 domain protects forks from MRE11-dependent resection and DSB conversion.","evidence":"SCAR-seq with separation-of-function mutants; auxin-degron depletion with DNA fiber analysis, EM of fork intermediates, and MRE11 inhibition","pmids":["30244834","30082684"],"confidence":"High","gaps":["Mechanism by which WD40 shields forks from MRE11 unknown","Coordination of histone-transfer and fork-protection roles undefined"]},{"year":2020,"claim":"Refined the assembly into a replication-factory model and resolved the human CMG docking interface, establishing trimer-mediated dimerization of two CMGs around one Pol alpha-primase.","evidence":"Cryo-EM of yeast Ctf4-CMG and 3.3 A human CMGA structures with in vitro reconstitution","pmids":["31589141","32453425"],"confidence":"High","gaps":["Functional importance of CMG dimerization in vivo not established","Identity of human-specific CMG recruitment factors still open"]},{"year":2022,"claim":"Extended the ATR-T826 phosphorylation mechanism to interstrand crosslinks, showing it triggers an intramolecular change that recruits FANCM/FAAP24 to activate Fanconi anemia signaling.","evidence":"T826A mutagenesis, ATR inhibition, Co-IP, proximity ligation, and FA pathway assays","pmids":["35867033"],"confidence":"High","gaps":["Structural nature of the intramolecular conformational change undefined","Whether the same switch governs both Claspin and FANCM recruitment unresolved"]},{"year":2024,"claim":"Placed WDHD1 within transcriptional regulatory and cancer contexts, identifying STAT3 and ZBTB16 as upstream regulators and proposing E3-ligase and CUL4B-coupled degradation behaviors.","evidence":"ChIP, ectopic expression/rescue, ubiquitination assays, and xenografts in lung adenocarcinoma models","pmids":["33413624","38654107","32426268","34923765","27099318"],"confidence":"Medium","gaps":["Claimed E3 ligase activity not reconstituted with purified components","Mechanistic link between transcriptional regulation and replisome function unclear"]},{"year":null,"claim":"It remains unresolved how the identity and ordering of human CMG recruitment factors, the competition among CIP-box partners at individual forks, and the coordination between WDHD1's replicative coupling and its damage-response/fork-protection roles are integrated in vivo.","evidence":"","pmids":[],"confidence":"Medium","gaps":["Human CMG recruitment route distinct from yeast GINS not molecularly identified","Hierarchy of competing CIP-box partners at a single fork unknown","Switch between replicative and repair functions not mechanistically defined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[0,11,20,25,26]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[7,16,17,23,34]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[11]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[17,34,36]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,9]},{"term_id":"GO:0000228","term_label":"nuclear chromosome","supporting_discovery_ids":[13,16,19]},{"term_id":"GO:0005654","term_label":"nucleoplasm","supporting_discovery_ids":[10,33]}],"pathway":[{"term_id":"R-HSA-69306","term_label":"DNA Replication","supporting_discovery_ids":[6,7,11,16,17]},{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[20,27,28,33,42]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[3,9,15]},{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[13,14,32]}],"complexes":["CMG (Cdc45-MCM2-7-GINS) helicase","replisome progression complex","Chl1-Ctf4-GINS assembly"],"partners":["POLA1","GINS (SLD5)","MCM10","RECQL4","DNA2","CHL1","CLSPN","FANCM"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O75717","full_name":"WD repeat and HMG-box DNA-binding protein 1","aliases":["Acidic nucleoplasmic DNA-binding protein 1","And-1"],"length_aa":1129,"mass_kda":126.0,"function":"Core replisome component that acts as a replication initiation factor. Binds directly to the CMG complex and functions as a hub to recruit additional proteins to the replication fork","subcellular_location":"Nucleus, nucleoplasm","url":"https://www.uniprot.org/uniprotkb/O75717/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/WDHD1","classification":"Common Essential","n_dependent_lines":1187,"n_total_lines":1208,"dependency_fraction":0.9826158940397351},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"SSRP1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/WDHD1","total_profiled":1310},"omim":[{"mim_id":"608126","title":"WD REPEAT AND HMG-BOX DNA-BINDING PROTEIN 1; WDHD1","url":"https://www.omim.org/entry/608126"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Nucleoplasm","reliability":"Enhanced"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"lymphoid tissue","ntpm":7.5}],"url":"https://www.proteinatlas.org/search/WDHD1"},"hgnc":{"alias_symbol":["AND-1","CTF4","CHTF4"],"prev_symbol":[]},"alphafold":{"accession":"O75717","domains":[{"cath_id":"2.130.10.10","chopping":"5-188_273-302","consensus_level":"high","plddt":92.5557,"start":5,"end":302},{"cath_id":"2.130.10.10","chopping":"427-620","consensus_level":"medium","plddt":95.7722,"start":427,"end":620},{"cath_id":"-","chopping":"771-829","consensus_level":"medium","plddt":92.4851,"start":771,"end":829},{"cath_id":"2.20.25","chopping":"626-717","consensus_level":"medium","plddt":96.0232,"start":626,"end":717}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O75717","model_url":"https://alphafold.ebi.ac.uk/files/AF-O75717-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O75717-F1-predicted_aligned_error_v6.png","plddt_mean":74.5},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=WDHD1","jax_strain_url":"https://www.jax.org/strain/search?query=WDHD1"},"sequence":{"accession":"O75717","fasta_url":"https://rest.uniprot.org/uniprotkb/O75717.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O75717/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O75717"}},"corpus_meta":[{"pmid":"15589699","id":"PMC_15589699","title":"Distribution of the vitamin D receptor and 1 alpha-hydroxylase in human brain.","date":"2005","source":"Journal of chemical neuroanatomy","url":"https://pubmed.ncbi.nlm.nih.gov/15589699","citation_count":1123,"is_preprint":false},{"pmid":"25912039","id":"PMC_25912039","title":"Vitamin D and 1,25(OH)2D regulation of T cells.","date":"2015","source":"Nutrients","url":"https://pubmed.ncbi.nlm.nih.gov/25912039","citation_count":385,"is_preprint":false},{"pmid":"11287619","id":"PMC_11287619","title":"Saccharomyces cerevisiae CTF18 and CTF4 are required for sister chromatid cohesion.","date":"2001","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/11287619","citation_count":272,"is_preprint":false},{"pmid":"12821938","id":"PMC_12821938","title":"VDUP1 upregulated by TGF-beta1 and 1,25-dihydorxyvitamin D3 inhibits tumor cell growth by blocking cell-cycle progression.","date":"2003","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/12821938","citation_count":234,"is_preprint":false},{"pmid":"19661920","id":"PMC_19661920","title":"A key role for Ctf4 in coupling the MCM2-7 helicase to DNA polymerase alpha within the eukaryotic replisome.","date":"2009","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/19661920","citation_count":215,"is_preprint":false},{"pmid":"16104866","id":"PMC_16104866","title":"Isolation and characterization of Dehalococcoides sp. strain FL2, a trichloroethene (TCE)- and 1,2-dichloroethene-respiring anaerobe.","date":"2005","source":"Environmental microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/16104866","citation_count":205,"is_preprint":false},{"pmid":"2175914","id":"PMC_2175914","title":"Vitamin D receptor interaction with specific DNA requires a nuclear protein and 1,25-dihydroxyvitamin D3.","date":"1990","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/2175914","citation_count":184,"is_preprint":false},{"pmid":"17761813","id":"PMC_17761813","title":"Mcm10 and And-1/CTF4 recruit DNA polymerase alpha to chromatin for initiation of DNA replication.","date":"2007","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/17761813","citation_count":180,"is_preprint":false},{"pmid":"24805245","id":"PMC_24805245","title":"A Ctf4 trimer couples the CMG helicase to DNA polymerase α in the eukaryotic replisome.","date":"2014","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/24805245","citation_count":177,"is_preprint":false},{"pmid":"30244834","id":"PMC_30244834","title":"The Mcm2-Ctf4-Polα Axis Facilitates Parental Histone H3-H4 Transfer to Lagging Strands.","date":"2018","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/30244834","citation_count":164,"is_preprint":false},{"pmid":"19805216","id":"PMC_19805216","title":"Assembly of the Cdc45-Mcm2-7-GINS complex in human cells requires the Ctf4/And-1, RecQL4, and Mcm10 proteins.","date":"2009","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/19805216","citation_count":162,"is_preprint":false},{"pmid":"15226378","id":"PMC_15226378","title":"Sister-chromatid cohesion mediated by the alternative RF-CCtf18/Dcc1/Ctf8, the helicase Chl1 and the polymerase-alpha-associated protein Ctf4 is essential for chromatid disjunction during meiosis II.","date":"2004","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/15226378","citation_count":126,"is_preprint":false},{"pmid":"22746383","id":"PMC_22746383","title":"The clinical toxicology of γ-hydroxybutyrate, γ-butyrolactone and 1,4-butanediol.","date":"2012","source":"Clinical toxicology (Philadelphia, Pa.)","url":"https://pubmed.ncbi.nlm.nih.gov/22746383","citation_count":126,"is_preprint":false},{"pmid":"25661486","id":"PMC_25661486","title":"Error-free DNA damage tolerance and sister chromatid proximity during DNA replication rely on the Polα/Primase/Ctf4 Complex.","date":"2015","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/25661486","citation_count":121,"is_preprint":false},{"pmid":"10101169","id":"PMC_10101169","title":"Dna2 mutants reveal interactions with Dna polymerase alpha and Ctf4, a Pol alpha accessory factor, and show that full Dna2 helicase activity is not essential for growth.","date":"1999","source":"Genetics","url":"https://pubmed.ncbi.nlm.nih.gov/10101169","citation_count":121,"is_preprint":false},{"pmid":"3031669","id":"PMC_3031669","title":"Pathway for inositol 1,3,4-trisphosphate and 1,4-bisphosphate metabolism.","date":"1987","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/3031669","citation_count":120,"is_preprint":false},{"pmid":"6253818","id":"PMC_6253818","title":"Leaky +1 and -1 frameshift mutations at the same site in a yeast mitochondrial gene.","date":"1980","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/6253818","citation_count":113,"is_preprint":false},{"pmid":"17073630","id":"PMC_17073630","title":"Recent advances in coumarins and 1-azacoumarins as versatile biodynamic agents.","date":"2006","source":"Current medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17073630","citation_count":110,"is_preprint":false},{"pmid":"27397686","id":"PMC_27397686","title":"Ctf4 Links DNA Replication with Sister Chromatid Cohesion Establishment by Recruiting the Chl1 Helicase to the Replisome.","date":"2016","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/27397686","citation_count":107,"is_preprint":false},{"pmid":"16348484","id":"PMC_16348484","title":"Degradation of 1,2,4-trichloro- and 1,2,4,5-tetrachlorobenzene by pseudomonas strains.","date":"1991","source":"Applied and environmental microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/16348484","citation_count":103,"is_preprint":false},{"pmid":"17299106","id":"PMC_17299106","title":"Calcium and 1,25-dihydroxyvitamin D3 regulation of adipokine expression.","date":"2007","source":"Obesity (Silver Spring, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/17299106","citation_count":102,"is_preprint":false},{"pmid":"12019279","id":"PMC_12019279","title":"Angiotensin 1-9 and 1-7 release in human heart: role of cathepsin A.","date":"2002","source":"Hypertension (Dallas, Tex. : 1979)","url":"https://pubmed.ncbi.nlm.nih.gov/12019279","citation_count":102,"is_preprint":false},{"pmid":"27397685","id":"PMC_27397685","title":"Ctf4 Is a Hub in the Eukaryotic Replisome that Links Multiple CIP-Box Proteins to the CMG Helicase.","date":"2016","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/27397685","citation_count":101,"is_preprint":false},{"pmid":"15576352","id":"PMC_15576352","title":"Repair of 3-methylthymine and 1-methylguanine lesions by bacterial and human AlkB proteins.","date":"2004","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/15576352","citation_count":95,"is_preprint":false},{"pmid":"15225800","id":"PMC_15225800","title":"Calcium and 1,25(OH)2D: interacting drivers of epidermal differentiation.","date":"2004","source":"The Journal of steroid biochemistry and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/15225800","citation_count":90,"is_preprint":false},{"pmid":"25079879","id":"PMC_25079879","title":"Are 1,4- and 1,5-disubstituted 1,2,3-triazoles good pharmacophoric groups?","date":"2014","source":"ChemMedChem","url":"https://pubmed.ncbi.nlm.nih.gov/25079879","citation_count":83,"is_preprint":false},{"pmid":"1341195","id":"PMC_1341195","title":"CTF4 (CHL15) mutants exhibit defective DNA metabolism in the yeast Saccharomyces cerevisiae.","date":"1992","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/1341195","citation_count":80,"is_preprint":false},{"pmid":"29123096","id":"PMC_29123096","title":"ATR kinase inhibition induces unscheduled origin firing through a Cdc7-dependent association between GINS and And-1.","date":"2017","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/29123096","citation_count":73,"is_preprint":false},{"pmid":"19496828","id":"PMC_19496828","title":"Ctf4 coordinates the progression of helicase and DNA polymerase alpha.","date":"2009","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/19496828","citation_count":72,"is_preprint":false},{"pmid":"32453425","id":"PMC_32453425","title":"CryoEM structures of human CMG-ATPγS-DNA and CMG-AND-1 complexes.","date":"2020","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/32453425","citation_count":71,"is_preprint":false},{"pmid":"25981845","id":"PMC_25981845","title":"Differential expression of plasma microRNA in preeclamptic patients at delivery and 1 year postpartum.","date":"2015","source":"American journal of obstetrics and gynecology","url":"https://pubmed.ncbi.nlm.nih.gov/25981845","citation_count":61,"is_preprint":false},{"pmid":"26110019","id":"PMC_26110019","title":"Metabolic pathway engineering for production of 1,2-propanediol and 1-propanol by Corynebacterium glutamicum.","date":"2015","source":"Biotechnology for biofuels","url":"https://pubmed.ncbi.nlm.nih.gov/26110019","citation_count":59,"is_preprint":false},{"pmid":"4469195","id":"PMC_4469195","title":"Morphological and biochemical effects of 1,2-dimethylhydrazine and 1-methylhydrazine in rats and mice.","date":"1974","source":"British journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/4469195","citation_count":57,"is_preprint":false},{"pmid":"20355099","id":"PMC_20355099","title":"Effect of ethylene and 1-MCP treatments on strawberry fruit ripening.","date":"2010","source":"Journal of the science of food and agriculture","url":"https://pubmed.ncbi.nlm.nih.gov/20355099","citation_count":55,"is_preprint":false},{"pmid":"20089864","id":"PMC_20089864","title":"Influence of the human cohesion establishment factor Ctf4/AND-1 on DNA replication.","date":"2010","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20089864","citation_count":54,"is_preprint":false},{"pmid":"28854333","id":"PMC_28854333","title":"Combining CRISPR and CRISPRi Systems for Metabolic Engineering of E. coli and 1,4-BDO Biosynthesis.","date":"2017","source":"ACS synthetic biology","url":"https://pubmed.ncbi.nlm.nih.gov/28854333","citation_count":54,"is_preprint":false},{"pmid":"31589141","id":"PMC_31589141","title":"Ctf4 organizes sister replisomes and Pol α into a replication factory.","date":"2019","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/31589141","citation_count":53,"is_preprint":false},{"pmid":"15371551","id":"PMC_15371551","title":"Predicting genes expressed via -1 and +1 frameshifts.","date":"2004","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/15371551","citation_count":50,"is_preprint":false},{"pmid":"12455694","id":"PMC_12455694","title":"mcl1+, the Schizosaccharomyces pombe homologue of CTF4, is important for chromosome replication, cohesion, and segregation.","date":"2002","source":"Eukaryotic cell","url":"https://pubmed.ncbi.nlm.nih.gov/12455694","citation_count":48,"is_preprint":false},{"pmid":"20627202","id":"PMC_20627202","title":"Biomonitoring of benzene and 1,3-butadiene exposure and early biological effects in traffic policemen.","date":"2010","source":"The Science of the total environment","url":"https://pubmed.ncbi.nlm.nih.gov/20627202","citation_count":47,"is_preprint":false},{"pmid":"30082684","id":"PMC_30082684","title":"AND-1 fork protection function prevents fork resection and is essential for proliferation.","date":"2018","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/30082684","citation_count":46,"is_preprint":false},{"pmid":"29167311","id":"PMC_29167311","title":"The human CTF4-orthologue AND-1 interacts with DNA polymerase α/primase via its unique C-terminal HMG box.","date":"2017","source":"Open biology","url":"https://pubmed.ncbi.nlm.nih.gov/29167311","citation_count":45,"is_preprint":false},{"pmid":"28525744","id":"PMC_28525744","title":"Ctf4 Prevents Genome Rearrangements by Suppressing DNA Double-Strand Break Formation and Its End Resection at Arrested Replication Forks.","date":"2017","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/28525744","citation_count":45,"is_preprint":false},{"pmid":"19439411","id":"PMC_19439411","title":"Roles of human AND-1 in chromosome transactions in S phase.","date":"2009","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/19439411","citation_count":45,"is_preprint":false},{"pmid":"31341109","id":"PMC_31341109","title":"Serine and 1-carbon metabolism are required for HIF-mediated protection against retinopathy of prematurity.","date":"2019","source":"JCI insight","url":"https://pubmed.ncbi.nlm.nih.gov/31341109","citation_count":45,"is_preprint":false},{"pmid":"21266480","id":"PMC_21266480","title":"WDHD1 modulates the post-transcriptional step of the centromeric silencing pathway.","date":"2011","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/21266480","citation_count":44,"is_preprint":false},{"pmid":"15354558","id":"PMC_15354558","title":"Enzyme immobilization on epoxy- and 1,1'-carbonyldiimidazole-activated methacrylate-based monoliths.","date":"2004","source":"Journal of separation science","url":"https://pubmed.ncbi.nlm.nih.gov/15354558","citation_count":44,"is_preprint":false},{"pmid":"27041081","id":"PMC_27041081","title":"Comparative effect of 25(OH)D3 and 1,25(OH)2D3 on Th17 cell differentiation.","date":"2016","source":"Clinical immunology (Orlando, Fla.)","url":"https://pubmed.ncbi.nlm.nih.gov/27041081","citation_count":44,"is_preprint":false},{"pmid":"3943146","id":"PMC_3943146","title":"Cytotoxic and mutagenic effects of 1-nitropyrene and 1-nitrosopyrene in diploid human fibroblasts.","date":"1986","source":"Carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/3943146","citation_count":44,"is_preprint":false},{"pmid":"24255107","id":"PMC_24255107","title":"Interaction between human Ctf4 and the Cdc45/Mcm2-7/GINS (CMG) replicative helicase.","date":"2013","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/24255107","citation_count":43,"is_preprint":false},{"pmid":"25756836","id":"PMC_25756836","title":"Interactions of arabinoxylan and (1,3)(1,4)-β-glucan with cellulose networks.","date":"2015","source":"Biomacromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/25756836","citation_count":43,"is_preprint":false},{"pmid":"30654237","id":"PMC_30654237","title":"Synthesis and anticancer evaluation of new lipophilic 1,2,4 and 1,3,4-oxadiazoles.","date":"2019","source":"European journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/30654237","citation_count":43,"is_preprint":false},{"pmid":"25490349","id":"PMC_25490349","title":"Systematically engineering Escherichia coli for enhanced production of 1,2-propanediol and 1-propanol.","date":"2014","source":"ACS synthetic biology","url":"https://pubmed.ncbi.nlm.nih.gov/25490349","citation_count":43,"is_preprint":false},{"pmid":"19429426","id":"PMC_19429426","title":"Antiproliferative action of menadione and 1,25(OH)2D3 on breast cancer cells.","date":"2009","source":"The Journal of steroid biochemistry and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/19429426","citation_count":42,"is_preprint":false},{"pmid":"20227510","id":"PMC_20227510","title":"Spiritual absence and 1-year mortality after hematopoietic stem cell transplant.","date":"2010","source":"Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation","url":"https://pubmed.ncbi.nlm.nih.gov/20227510","citation_count":41,"is_preprint":false},{"pmid":"28173138","id":"PMC_28173138","title":"Synaptotagmin-2, and -1, linked to neurotransmission impairment and vulnerability in Spinal Muscular Atrophy.","date":"2016","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/28173138","citation_count":40,"is_preprint":false},{"pmid":"32075754","id":"PMC_32075754","title":"Sen1 Is Recruited to Replication Forks via Ctf4 and Mrc1 and Promotes Genome Stability.","date":"2020","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/32075754","citation_count":39,"is_preprint":false},{"pmid":"20237432","id":"PMC_20237432","title":"Xenopus DNA2 is a helicase/nuclease that is found in complexes with replication proteins And-1/Ctf4 and Mcm10 and DSB response proteins Nbs1 and ATM.","date":"2010","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/20237432","citation_count":39,"is_preprint":false},{"pmid":"31291500","id":"PMC_31291500","title":"Reactions of 2-Aryl-1,3-Dithianes and [1.1.1]Propellane.","date":"2019","source":"Angewandte Chemie (International ed. in English)","url":"https://pubmed.ncbi.nlm.nih.gov/31291500","citation_count":38,"is_preprint":false},{"pmid":"35959252","id":"PMC_35959252","title":"Antibacterial interactions of pulegone and 1,8-cineole with monolaurin ornisin against Staphylococcus aureus.","date":"2022","source":"Food science & nutrition","url":"https://pubmed.ncbi.nlm.nih.gov/35959252","citation_count":38,"is_preprint":false},{"pmid":"10926169","id":"PMC_10926169","title":"The role of calbindin and 1,25dihydroxyvitamin D3 in the kidney.","date":"2000","source":"Current opinion in nephrology and hypertension","url":"https://pubmed.ncbi.nlm.nih.gov/10926169","citation_count":36,"is_preprint":false},{"pmid":"34063148","id":"PMC_34063148","title":"Protective Role of Vanillic Acid against Diethylnitrosamine- and 1,2-Dimethylhydrazine-Induced Hepatocarcinogenesis in Rats.","date":"2021","source":"Molecules (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/34063148","citation_count":35,"is_preprint":false},{"pmid":"21725360","id":"PMC_21725360","title":"And-1 is required for the stability of histone acetyltransferase Gcn5.","date":"2011","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/21725360","citation_count":34,"is_preprint":false},{"pmid":"9175701","id":"PMC_9175701","title":"AND-1, a natural chimeric DNA-binding protein, combines an HMG-box with regulatory WD-repeats.","date":"1997","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/9175701","citation_count":32,"is_preprint":false},{"pmid":"25602958","id":"PMC_25602958","title":"RecQL4 is required for the association of Mcm10 and Ctf4 with replication origins in human cells.","date":"2015","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/25602958","citation_count":32,"is_preprint":false},{"pmid":"22374673","id":"PMC_22374673","title":"Mimosine arrests the cell cycle prior to the onset of DNA replication by preventing the binding of human Ctf4/And-1 to chromatin via Hif-1α activation in HeLa cells.","date":"2012","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/22374673","citation_count":32,"is_preprint":false},{"pmid":"28381552","id":"PMC_28381552","title":"The structure and polymerase-recognition mechanism of the crucial adaptor protein AND-1 in the human replisome.","date":"2017","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/28381552","citation_count":31,"is_preprint":false},{"pmid":"26082189","id":"PMC_26082189","title":"And-1 coordinates with Claspin for efficient Chk1 activation in response to replication stress.","date":"2015","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/26082189","citation_count":30,"is_preprint":false},{"pmid":"31004818","id":"PMC_31004818","title":"Bioelectrochemical assisted dechlorination of tetrachloroethylene and 1,2-dichloroethane by acclimation of anaerobic sludge.","date":"2019","source":"Chemosphere","url":"https://pubmed.ncbi.nlm.nih.gov/31004818","citation_count":30,"is_preprint":false},{"pmid":"30979824","id":"PMC_30979824","title":"Human CST suppresses origin licensing and promotes AND-1/Ctf4 chromatin association.","date":"2019","source":"Life science alliance","url":"https://pubmed.ncbi.nlm.nih.gov/30979824","citation_count":29,"is_preprint":false},{"pmid":"31421253","id":"PMC_31421253","title":"Design, synthesis and anticancer evaluation of novel 1,3-benzodioxoles and 1,4-benzodioxines.","date":"2019","source":"European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences","url":"https://pubmed.ncbi.nlm.nih.gov/31421253","citation_count":29,"is_preprint":false},{"pmid":"33413624","id":"PMC_33413624","title":"STAT3 plays an important role in DNA replication by turning on WDHD1.","date":"2021","source":"Cell & bioscience","url":"https://pubmed.ncbi.nlm.nih.gov/33413624","citation_count":28,"is_preprint":false},{"pmid":"27940552","id":"PMC_27940552","title":"And-1 coordinates with CtIP for efficient homologous recombination and DNA damage checkpoint maintenance.","date":"2017","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/27940552","citation_count":28,"is_preprint":false},{"pmid":"26645455","id":"PMC_26645455","title":"Cooperative Protein Folding by Two Protein Thiol Disulfide Oxidoreductases and 1 in Soybean.","date":"2015","source":"Plant physiology","url":"https://pubmed.ncbi.nlm.nih.gov/26645455","citation_count":27,"is_preprint":false},{"pmid":"28256373","id":"PMC_28256373","title":"Synthesis and biological evaluation of chalcone, dihydrochalcone, and 1,3-diarylpropane analogs as anti-inflammatory agents.","date":"2017","source":"Bioorganic & medicinal chemistry letters","url":"https://pubmed.ncbi.nlm.nih.gov/28256373","citation_count":26,"is_preprint":false},{"pmid":"24001290","id":"PMC_24001290","title":"GHB, GBL and 1,4-BD addiction.","date":"2014","source":"Current pharmaceutical design","url":"https://pubmed.ncbi.nlm.nih.gov/24001290","citation_count":25,"is_preprint":false},{"pmid":"27940557","id":"PMC_27940557","title":"And-1 is required for homologous recombination repair by regulating DNA end resection.","date":"2017","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/27940557","citation_count":25,"is_preprint":false},{"pmid":"2979739","id":"PMC_2979739","title":"Metabolism and DNA binding of 1-nitropyrene and 1-nitrosopyrene in newborn mice.","date":"1988","source":"Chemical research in toxicology","url":"https://pubmed.ncbi.nlm.nih.gov/2979739","citation_count":24,"is_preprint":false},{"pmid":"35239320","id":"PMC_35239320","title":"Geobacter sp. Strain IAE Dihaloeliminates 1,1,2-Trichloroethane and 1,2-Dichloroethane.","date":"2022","source":"Environmental science & technology","url":"https://pubmed.ncbi.nlm.nih.gov/35239320","citation_count":23,"is_preprint":false},{"pmid":"27099318","id":"PMC_27099318","title":"Role of WDHD1 in Human Papillomavirus-Mediated Oncogenesis Identified by Transcriptional Profiling of E7-Expressing Cells.","date":"2016","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/27099318","citation_count":22,"is_preprint":false},{"pmid":"32426268","id":"PMC_32426268","title":"WDHD1 Leads to Cisplatin Resistance by Promoting MAPRE2 Ubiquitination in Lung Adenocarcinoma.","date":"2020","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/32426268","citation_count":22,"is_preprint":false},{"pmid":"31006829","id":"PMC_31006829","title":"Co-treatment with interferon-γ and 1-methyl tryptophan ameliorates cardiac fibrosis through cardiac myofibroblasts apoptosis.","date":"2019","source":"Molecular and cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/31006829","citation_count":22,"is_preprint":false},{"pmid":"17222391","id":"PMC_17222391","title":"Chl1 and Ctf4 are required for damage-induced recombinations.","date":"2007","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/17222391","citation_count":21,"is_preprint":false},{"pmid":"28815832","id":"PMC_28815832","title":"Targeting the Genome-Stability Hub Ctf4 by Stapled-Peptide Design.","date":"2017","source":"Angewandte Chemie (International ed. in English)","url":"https://pubmed.ncbi.nlm.nih.gov/28815832","citation_count":20,"is_preprint":false},{"pmid":"32379761","id":"PMC_32379761","title":"Separable, Ctf4-mediated recruitment of DNA Polymerase α for initiation of DNA synthesis at replication origins and lagging-strand priming during replication elongation.","date":"2020","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/32379761","citation_count":20,"is_preprint":false},{"pmid":"33221821","id":"PMC_33221821","title":"WDHD1 is essential for the survival of PTEN-inactive triple-negative breast cancer.","date":"2020","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/33221821","citation_count":19,"is_preprint":false},{"pmid":"21470422","id":"PMC_21470422","title":"Drosophila Ctf4 is essential for efficient DNA replication and normal cell cycle progression.","date":"2011","source":"BMC molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/21470422","citation_count":19,"is_preprint":false},{"pmid":"25697176","id":"PMC_25697176","title":"Replisome function during replicative stress is modulated by histone h3 lysine 56 acetylation through Ctf4.","date":"2015","source":"Genetics","url":"https://pubmed.ncbi.nlm.nih.gov/25697176","citation_count":19,"is_preprint":false},{"pmid":"23277051","id":"PMC_23277051","title":"Suppressors of cytokine signalling-3 and -1 in human carcinogenesis.","date":"2013","source":"Frontiers in bioscience (Scholar edition)","url":"https://pubmed.ncbi.nlm.nih.gov/23277051","citation_count":18,"is_preprint":false},{"pmid":"9373193","id":"PMC_9373193","title":"Differentiation of myeloid cells and 1,25-dihydroxyvitamin D3.","date":"1997","source":"Leukemia & lymphoma","url":"https://pubmed.ncbi.nlm.nih.gov/9373193","citation_count":18,"is_preprint":false},{"pmid":"31353794","id":"PMC_31353794","title":"Catalytic Enantioselective Synthesis of 1,4-Keto-Alkenylboronate Esters and 1,4-Dicarbonyls.","date":"2019","source":"Angewandte Chemie (International ed. in English)","url":"https://pubmed.ncbi.nlm.nih.gov/31353794","citation_count":18,"is_preprint":false},{"pmid":"2066996","id":"PMC_2066996","title":"Purine and 1-deazapurine ribonucleosides and deoxyribonucleosides: synthesis and biological activity.","date":"1991","source":"Journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/2066996","citation_count":18,"is_preprint":false},{"pmid":"32061136","id":"PMC_32061136","title":"Immune epigenetic age in pregnancy and 1 year after birth: Associations with weight change.","date":"2020","source":"American journal of reproductive immunology (New York, N.Y. : 1989)","url":"https://pubmed.ncbi.nlm.nih.gov/32061136","citation_count":18,"is_preprint":false},{"pmid":"35867033","id":"PMC_35867033","title":"And-1 Coordinates with the FANCM Complex to Regulate Fanconi Anemia Signaling and Cisplatin Resistance.","date":"2022","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/35867033","citation_count":17,"is_preprint":false},{"pmid":"8806421","id":"PMC_8806421","title":"Characterization and expression of chitinase and 1,3-beta-glucanase genes in cotton.","date":"1996","source":"Plant molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/8806421","citation_count":17,"is_preprint":false},{"pmid":"15707965","id":"PMC_15707965","title":"Tyrosinase-related protein-2 and -1 are trafficked on distinct routes in B16 melanoma cells.","date":"2005","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/15707965","citation_count":16,"is_preprint":false},{"pmid":"26247835","id":"PMC_26247835","title":"Oxidation of pyrene, 1-hydroxypyrene, 1-nitropyrene and 1-acetylpyrene by human cytochrome P450 2A13.","date":"2015","source":"Xenobiotica; the fate of foreign compounds in biological systems","url":"https://pubmed.ncbi.nlm.nih.gov/26247835","citation_count":15,"is_preprint":false},{"pmid":"23973249","id":"PMC_23973249","title":"Serotonergic genes and suicidal ideation 2 weeks and 1 year after stroke in Korea.","date":"2013","source":"The American journal of geriatric psychiatry : official journal of the American Association for Geriatric Psychiatry","url":"https://pubmed.ncbi.nlm.nih.gov/23973249","citation_count":15,"is_preprint":false},{"pmid":"38654107","id":"PMC_38654107","title":"ZBTB16 inhibits DNA replication and induces cell cycle arrest by targeting WDHD1 transcription in lung adenocarcinoma.","date":"2024","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/38654107","citation_count":13,"is_preprint":false},{"pmid":"34923765","id":"PMC_34923765","title":"Discovery and characterization of potent And-1 inhibitors for cancer treatment.","date":"2021","source":"Clinical and translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34923765","citation_count":13,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":48373,"output_tokens":10450,"usd":0.150934,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":21019,"output_tokens":4790,"usd":0.112422,"stage2_stop_reason":"end_turn"},"total_usd":0.263356,"stage1_batch_id":"msgbatch_01UT1AahFPCAyn2uVweS7A9z","stage2_batch_id":"msgbatch_01HuKWtKNT4Pqj1bc8eEP5RV","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1997,\n      \"finding\": \"AND-1 (WDHD1) was identified and characterized as a novel nuclear DNA-binding protein containing an N-terminal WD-repeat domain and a C-terminal HMG-box domain. It binds four-way junction DNA, exists as an oligomer (likely homodimer), and localizes to the interchromatinic space during interphase but disperses to the cytoplasm during mitosis.\",\n      \"method\": \"cDNA cloning, monoclonal antibody characterization, DNA affinity chromatography, EMSA, immunolocalization\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal biochemical methods in a single study establishing domain architecture, DNA binding, and localization\",\n      \"pmids\": [\"9175701\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1992,\n      \"finding\": \"CTF4/CHL15 (yeast ortholog of WDHD1) encodes a ~105 kDa protein that associates in vitro with DNA polymerase alpha; ctf4 mutants show elevated sister chromatid loss and recombination but not elevated point mutation rates, and the C-terminal helix-loop-helix domain is essential for CTF4 function.\",\n      \"method\": \"Yeast genetics, chromosome segregation assays, in vitro DNA polymerase alpha association, sequence analysis, frameshift mutagenesis\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic and biochemical methods in a single lab establishing Pol alpha association and functional domain requirements\",\n      \"pmids\": [\"1341195\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"Yeast CTF4 genetically interacts with DNA2 and POL1 (Pol alpha subunit); ctf4 deletion is synthetically lethal with DNA damage-sensitive dna2 alleles, placing Ctf4 in a pathway with Dna2 and Pol alpha for lagging strand synthesis or repair.\",\n      \"method\": \"Yeast genetic epistasis, synthetic lethality analysis, allele construction\",\n      \"journal\": \"Genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with multiple alleles establishing pathway position\",\n      \"pmids\": [\"10101169\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"Yeast CTF4 is required for sister chromatid cohesion; ctf4 null mutants exhibit cohesion failure and preanaphase accumulation dependent on the spindle assembly checkpoint, placing CTF4 functionally at the replication fork in the cohesion establishment pathway.\",\n      \"method\": \"Yeast genetics, sister chromatid cohesion assay, checkpoint epistasis\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — replicated across labs, cohesion assay with checkpoint epistasis establishing pathway position\",\n      \"pmids\": [\"11287619\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Fission yeast Mcl1 (ortholog of CTF4/WDHD1) is required for chromosome replication, sister chromatid cohesion, and segregation; mcl1-1 is synthetically lethal with checkpoint mutations rad3Δ and rad26Δ, and interacts genetically with replication components.\",\n      \"method\": \"Fission yeast genetics, pulsed-field electrophoresis, synthetic lethality, overexpression analysis\",\n      \"journal\": \"Eukaryotic cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis and cell biological phenotypes in a single lab\",\n      \"pmids\": [\"12455694\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Yeast CTF4 (polymerase-alpha-associated protein) is required for efficient sister chromatid cohesion in both mitosis and meiosis; ctf4Δ cells show massive meiosis II non-disjunction and loss of centromeric cohesion despite normal cohesin association with centromeric DNA.\",\n      \"method\": \"Yeast genetics, cohesion assays, cytological analysis of meiosis\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple phenotypic assays in a single lab establishing meiotic cohesion requirement\",\n      \"pmids\": [\"15226378\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Human AND-1/CTF4 (WDHD1) interacts with Mcm10 (which associates with MCM2-7) and with the p180 subunit of DNA polymerase alpha. In Xenopus egg extracts, AND-1 loads on chromatin after Mcm10, concurrently with Pol alpha, and is required for efficient DNA synthesis. Mcm10 is required for chromatin loading of AND-1 and Pol alpha; disrupting the Mcm10-AND-1 interaction inhibits DNA synthesis.\",\n      \"method\": \"Co-immunoprecipitation, Xenopus egg extract DNA replication assay, chromatin fractionation, inhibitory antibody, siRNA knockdown in mammalian cells\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (Co-IP, antibody inhibition, egg extract reconstitution, siRNA) across human and Xenopus systems\",\n      \"pmids\": [\"17761813\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Yeast Ctf4 couples the MCM2-7 helicase to DNA polymerase alpha within the replisome progression complex (RPC); Ctf4 and GINS together are crucial for this coupling. In ctf4Δ cells, Pol alpha is unstable and barely detectable at replication forks, while helicase (and Pol epsilon) quantities are unchanged but their localizations are abnormally dispersed. Cells lacking both Ctf4 and Mrc1 experience chronic DNA damage checkpoint activation and fail to complete the cell cycle.\",\n      \"method\": \"Yeast genetics, ChIP, in vitro interaction assays between GINS and Pol alpha mediated by Ctf4, immunoprecipitation, double mutant analysis\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — ChIP, in vitro reconstitution of GINS-Ctf4-Pol alpha interaction, and genetic double-mutant analysis in a single study\",\n      \"pmids\": [\"19661920\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Yeast Ctf4 mediates the in vitro interaction between GINS and DNA Pol alpha-primase; in ctf4Δ mutants, the MCM-GINS–Pol alpha interaction is specifically destabilized, Pol alpha becomes unstable at replication forks, and the replication checkpoint is activated. Tof1 and Mrc1 are not required for this interaction.\",\n      \"method\": \"Yeast genetics, in vitro protein interaction assay, ChIP, immunoblot\",\n      \"journal\": \"Genes to cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro interaction plus ChIP in a single lab\",\n      \"pmids\": [\"19496828\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Human AND-1/CTF4 (WDHD1) is required for assembly of the CMG (Cdc45-Mcm2-7-GINS) complex in human cells; along with RecQL4 and Mcm10, AND-1 is needed for stable CMG interactions observed only after G1/S transition and dependent on CDK and Cdc7 kinase activities.\",\n      \"method\": \"Bimolecular fluorescence complementation (BiFC) in HeLa cells, siRNA depletion, CDK and Cdc7 kinase inhibition\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — BiFC assay with siRNA and kinase inhibitor controls in a single lab\",\n      \"pmids\": [\"19805216\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Human AND-1 (WDHD1) localizes adjacent to replication foci in S phase, is phosphorylated in response to replication arrest in a checkpoint kinase (ATM, ATR, Cdc7)-dependent manner, interacts with cohesin proteins Smc1, Smc3, and Rad21/Scc1, and is required for homologous recombination repair. Depletion of AND-1 increases DNA damage, delays S-phase progression, compromises checkpoint activation, and decreases Chk1 protein levels.\",\n      \"method\": \"Immunofluorescence, co-immunoprecipitation, siRNA knockdown, HR repair assay (I-SceI DSB), immunoblot, flow cytometry\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods in a single lab establishing localization and interaction with cohesin\",\n      \"pmids\": [\"19439411\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Purified human CTF4 (WDHD1) exists as a dimer; the SepB domain determines dimeric structure. hCtf4 binds preferentially to DNA template-primer structures, directly interacts with DNA polymerases alpha, delta, and epsilon, and markedly stimulates Pol alpha and Pol epsilon activities in vitro. siRNA depletion of hCtf4 in HeLa cells causes G1/S arrest and slower DNA replication.\",\n      \"method\": \"Protein purification, in vitro DNA polymerase stimulation assay, EMSA/DNA binding, co-immunoprecipitation, siRNA knockdown, DNA fiber analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro biochemical reconstitution of Pol stimulation with purified proteins, plus multiple orthogonal methods\",\n      \"pmids\": [\"20089864\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Xenopus Dna2 forms a complex with And-1/Ctf4 and Mcm10 at replication forks (demonstrated by co-immunoprecipitation from egg extracts), and is recruited to DNA after replication origin licensing.\",\n      \"method\": \"Xenopus egg extract, co-immunoprecipitation, immunofluorescence/co-localization\",\n      \"journal\": \"Cell cycle\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-IP in egg extracts, replicated in context of a broader study\",\n      \"pmids\": [\"20237432\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Human WDHD1 associates with centromeres in a cell cycle-dependent manner (mid-to-late S phase), and its down-regulation compromises HP1alpha localization to pericentric heterochromatin, alters epigenetic marks at pericentric chromatin, disrupts mitosis, and impairs generation of centromeric satellite small non-coding RNAs by reducing Dicer association with centromeric RNA.\",\n      \"method\": \"ChIP, immunofluorescence, siRNA knockdown, chromatin fractionation, RNA analysis\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (ChIP, IF, RNA analysis) in a single lab\",\n      \"pmids\": [\"21266480\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"AND-1 (WDHD1) forms a complex with both histone H3 and histone acetyltransferase Gcn5; AND-1 down-regulation causes Gcn5 protein degradation, reducing H3K9 and H3K56 acetylation. AND-1 overexpression stabilizes Gcn5 through protein-protein interaction.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, immunoblot, overexpression\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP and knockdown/overexpression in a single lab; no reconstitution with purified proteins\",\n      \"pmids\": [\"21725360\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Mimosine prevents chromatin binding of Ctf4/AND-1 in HeLa cells, arresting the cell cycle in G1 before S-phase entry via a Hif-1alpha-dependent increase in p27; depletion of Hif-1alpha restores Ctf4 chromatin binding and allows S-phase entry even in the presence of mimosine.\",\n      \"method\": \"Chromatin fractionation, siRNA knockdown, cell cycle analysis, flow cytometry\",\n      \"journal\": \"Cell cycle\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — chromatin binding assay plus siRNA rescue establishing pathway\",\n      \"pmids\": [\"22374673\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Human CTF4 (WDHD1) interacts with the CMG (Cdc45-Mcm2-7-GINS) complex; the hCtf4-CMG complex was isolated by in vitro interaction of purified proteins, by co-infection of insect cells, and from HeLa cell chromatin. The complex contains homodimeric hCtf4 and monomeric CMG. The hCtf4-CMG complex retains DNA helicase activity that is more salt-resistant than CMG alone. The stability of hCtf4-CMG depends on interactions between hCtf4 and multiple CMG components.\",\n      \"method\": \"Purified protein interaction, Sf9 co-infection/co-purification, HeLa chromatin immunoprecipitation, helicase assay\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — reconstituted with purified proteins, validated in insect cells and from human chromatin, enzymatic assay\",\n      \"pmids\": [\"24255107\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Yeast Ctf4 self-associates as a constitutive disk-shaped homotrimer via a C-terminal beta-propeller (SepB) domain fused to a helical extension. Pol alpha catalytic subunit and GINS subunit Sld5 share a conserved Ctf4-interacting peptide (CIP-box) that docks onto the helical extension of a Ctf4 protomer; one Ctf4 trimer can simultaneously bind both Pol alpha and GINS, enabling coupling of two Pol alpha molecules to one CMG helicase.\",\n      \"method\": \"X-ray crystallography, electron microscopy, native mass spectrometry, mutagenesis, in vitro binding assays\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure and EM with functional mutagenesis and native MS, multiple orthogonal methods in a rigorous single study\",\n      \"pmids\": [\"24805245\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Yeast Pol alpha/Primase/Ctf4 mutants that are proficient in bulk DNA replication are defective in recombination-mediated damage bypass by template switching and have reduced sister chromatid cohesion; these DDT defects are caused by altered ssDNA metabolism and abnormal replication fork topology rather than increased sister chromatid distance.\",\n      \"method\": \"Yeast genetics, DNA fiber analysis, electron microscopy of replication intermediates, sister chromatid cohesion assay\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (genetics, EM of fork structures, fiber analysis) establishing mechanistic distinction\",\n      \"pmids\": [\"25661486\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"RecQL4 is required for the association of Mcm10 and Ctf4 with replication origins in human cells; physical interactions between RecQL4, Mcm10, and Ctf4 and their origin association require both CDK and DDK activities and are blocked by the DNA damage checkpoint.\",\n      \"method\": \"ChIP at replication origins, co-immunoprecipitation, siRNA knockdown, kinase inhibition\",\n      \"journal\": \"Cell cycle\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP and Co-IP with kinase inhibitor controls in a single lab\",\n      \"pmids\": [\"25602958\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"In response to replication stress, AND-1 is phosphorylated at T826 by ATR; this phosphorylation causes AND-1 to accumulate at damage sites where it promotes the Claspin-Chk1 interaction, stimulating efficient Chk1 activation by ATR. AND-1 directly binds ssDNA and facilitates Claspin association with ssDNA. AND-1 is required for recovery of stalled forks.\",\n      \"method\": \"Phosphorylation site mutagenesis, co-immunoprecipitation, in vitro ssDNA binding assay, siRNA knockdown, immunofluorescence, DNA fiber analysis\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — phospho-site mutagenesis combined with Co-IP, in vitro ssDNA binding, and functional rescue in a single study\",\n      \"pmids\": [\"26082189\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Histone H3K56 acetylation acts through Ctf4 to uncouple CMG helicase from DNA polymerases under replication stress; the N-terminal domain of Ctf4 interacts with Mms22 (adaptor of the Rtt101-Mms1 E3 ubiquitin ligase), and this interaction is promoted by replicative stress.\",\n      \"method\": \"Yeast genetics, genetic epistasis, domain deletion analysis, two-hybrid/co-immunoprecipitation\",\n      \"journal\": \"Genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis and domain analysis in a single lab\",\n      \"pmids\": [\"25697176\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Ctf4 recruits the Chl1 helicase to the replisome via a conserved CIP-box interaction motif shared with GINS and Pol alpha; EM analysis visualized a reconstituted Chl1-Ctf4-GINS assembly. Ctf4 interaction (but not Chl1 helicase activity) is required for sister chromatid cohesion, while Chl1 helicase activity is required for replication fork progression under nucleotide depletion. Physical interaction between Chl1 and cohesin occurs during S phase.\",\n      \"method\": \"Electron microscopy of reconstituted complex, in vitro binding assays, yeast genetics with separation-of-function mutants, co-immunoprecipitation\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — EM-validated reconstituted complex, in vitro binding, separation-of-function genetics, and Co-IP across multiple methods\",\n      \"pmids\": [\"27397686\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Ctf4 acts as a hub connecting multiple CIP-box proteins (in addition to Pol alpha and GINS) to the CMG helicase; crystallographic analysis classifies CIP-boxes into two groups targeting different sites on Ctf4. Ctf4-dependent recruitment of Dna2 and Tof2 via their CIP-boxes is required for rDNA copy-number regulation but not genome-wide DNA synthesis.\",\n      \"method\": \"X-ray crystallography, in vitro binding assays, yeast genetics, CIP-box mutagenesis, genome-wide replication analysis\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structures with functional mutagenesis and genetics establishing multi-partner hub mechanism\",\n      \"pmids\": [\"27397685\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"WDHD1 (AND-1) plays an important role in E7 (HPV-16)-induced G1 checkpoint abrogation and rereplication; WDHD1 protein steady-state level and half-life are increased in E7-expressing cells, and WDHD1 knockdown reduces E7-induced G1 checkpoint abrogation.\",\n      \"method\": \"siRNA knockdown, immunoblot (protein stability), cell cycle analysis, rereplication assay\",\n      \"journal\": \"Journal of virology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with specific cell cycle phenotype in a single lab\",\n      \"pmids\": [\"27099318\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Human AND-1 (WDHD1) maintains the trimeric structure of yeast Ctf4 via its conserved SepB domain, but interacts with DNA Pol alpha/primase primarily through its C-terminal HMG box, which binds the Pol alpha B subunit at the same site targeted by SV40 T-antigen. AND-1 also possesses a DNA-binding activity via its HMG box that may position Pol alpha on the lagging-strand template.\",\n      \"method\": \"Biochemical protein interaction assays, domain mutagenesis, competition with SV40 T-antigen peptide, DNA binding assay\",\n      \"journal\": \"Open biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro biochemical reconstitution with mutagenesis and competition experiments establishing interaction mechanism\",\n      \"pmids\": [\"29167311\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Human AND-1 (WDHD1) exists as a homotrimer mediated by the SepB domain; the positively charged groove within SepB provides binding sites for Pol alpha. AND-1 binds DNA in vitro via its C-terminal HMG domain. Unlike yeast Ctf4, human AND-1's recruitment to the CMG complex appears to be mediated by unknown participants other than GINS.\",\n      \"method\": \"X-ray crystallography of WD40 and SepB domains, biochemical assays, mutagenesis, DNA binding assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure with biochemical validation and mutagenesis in a single study\",\n      \"pmids\": [\"28381552\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"AND-1/Ctf4 promotes DNA end resection and homologous recombination repair by interacting with CtIP and regulating its recruitment to DNA damage sites. AND-1 localizes to damage sites via the MDC1-RNF8 pathway and is required for sustained ATR-CHK1 signaling and maintenance of intra-S and G2-phase checkpoints.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, immunofluorescence at damage foci, HR repair assay, checkpoint assays\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple methods (Co-IP, IF, functional assays) in a single lab\",\n      \"pmids\": [\"27940552\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"AND-1 (WDHD1) forms complexes with CtIP and other repair proteins; AND-1 is recruited to DSB sites dependent on MDC1, BRCA1, and ATM. AND-1 knockdown impairs DSB end resection by reducing CtIP recruitment, reduces Chk1 activation, establishing an MDC1→AND-1→CtIP axis for CtIP-mediated end resection.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, immunofluorescence at damage foci, end resection assay, Chk1 activation assay\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple methods in a single lab; replicates the Chen et al. 2017 finding from a different group\",\n      \"pmids\": [\"27940557\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Yeast Ctf4 prevents genome rearrangements by suppressing DSB formation and end resection at arrested replication forks; in ctf4Δ cells, DSBs form more frequently at arrested forks, undergo end resection, and are repaired by HR prone to rDNA hyper-amplification.\",\n      \"method\": \"Yeast genetics, Southern blotting/rDNA copy number, DSB quantification, end resection assay\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic and molecular analysis of DSB end resection with multiple assays in a single lab\",\n      \"pmids\": [\"28525744\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"A stapled peptide encoding the Ctf4-interacting peptide (CIP) of Sld5 binds Ctf4 with submicromolar affinity; crystal structure confirmed the mode of interaction; the stapled peptide displaces Pol alpha from the replisome in yeast extracts.\",\n      \"method\": \"Stapled peptide design, crystal structure, fluorescence polarization binding assay, yeast extract replisome disruption assay\",\n      \"journal\": \"Angewandte Chemie\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure plus functional displacement assay with purified/extract proteins\",\n      \"pmids\": [\"28815832\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"ATR-Chk1 inhibitor-induced unscheduled origin firing in human cells is mediated by Cdc7 kinase through phosphorylations on GINS that induce an association between GINS and AND-1 (WDHD1).\",\n      \"method\": \"Proteomics (phosphoproteomics), co-immunoprecipitation, kinase inhibition, siRNA knockdown\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — phosphoproteomic identification with Co-IP validation and kinase inhibitor evidence in a single lab\",\n      \"pmids\": [\"29123096\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"The Mcm2-Ctf4-Pol alpha axis facilitates transfer of parental histone (H3-H4)2 tetramers to lagging-strand DNA at replication forks; mutations disrupting the Mcm2 histone-binding domain, or Ctf4-Pol alpha connection, result in enrichment of parental histones on leading strand DNA.\",\n      \"method\": \"SCAR-seq (strand-specific parental histone enrichment), yeast genetics with separation-of-function mutations, ChIP\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — strand-specific histone tracking with multiple separation-of-function mutants across Mcm2, Ctf4, and Pol alpha in a single rigorous study\",\n      \"pmids\": [\"30244834\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"AND-1 (WDHD1) protects replication forks from resection; depletion of AND-1 using an inducible degron causes fork speed slow-down, accumulation of long ssDNA gaps at the fork junction, and conversion of these to DSBs in G2 via MRE11 nuclease activity. The WD40 domain of AND-1 is specifically required for fork protection and proliferation, while the HMG box is required for fast replication but not proliferation.\",\n      \"method\": \"Auxin-inducible degron (AND-1 depletion), DNA fiber analysis, electron microscopy of replication intermediates, MRE11 inhibitor (mirin), domain-specific mutants\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — inducible degron system with domain separation-of-function, EM of fork structures, and pharmacological validation in a single rigorous study\",\n      \"pmids\": [\"30082684\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Ctf4 tightly dimerizes CMG helicase via an interface involving Psf2, Cdc45, and Sld5; Ctf4 binds only one Pol alpha-primase per trimer, suggesting a replication factory model where two CMG helicases and one Pol alpha-primase are organized by one Ctf4 trimer. Cryo-EM reveals two CMGs nearly facing each other with lagging strands toward the center.\",\n      \"method\": \"Cryo-EM structure, in vitro reconstitution, pulldown assays\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — cryo-EM structure with in vitro reconstitution establishing stoichiometry and architecture\",\n      \"pmids\": [\"31589141\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Human CST (CTC1-STN1-TEN1) promotes AND-1/Ctf4 chromatin association and replisome assembly; CST directly interacts with AND-1 and pol alpha, enhancing their chromatin association independently of exogenous replication stress.\",\n      \"method\": \"Co-immunoprecipitation, chromatin fractionation, siRNA knockdown, origin licensing assays\",\n      \"journal\": \"Life science alliance\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP with chromatin fractionation in a single lab\",\n      \"pmids\": [\"30979824\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"CryoEM structure of human CMG bound to AND-1 (CMGA) reveals the AND-1 trimer docks onto the helicase assembly formed by Cdc45 and GINS via one beta-propeller domain of its trimerization region; in the CMGA architecture, AND-1 is closely positioned to fork DNA while its CIP-binding helical domains remain available for partner protein recruitment.\",\n      \"method\": \"CryoEM structure at 3.3 Å resolution\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — high-resolution cryoEM structure establishing the molecular docking interface\",\n      \"pmids\": [\"32453425\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Yeast Sen1 (transcription termination helicase) associates with the replisome via Ctf4 and Mrc1; the N-terminus of Sen1 binds Ctf4 and Mrc1. A separation-of-function mutant (sen1-3) that abolishes replisome binding without affecting transcription termination shows increased genome instability and recombination.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation, separation-of-function mutagenesis, genetics\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — separation-of-function mutant with Co-IP validation in a single lab\",\n      \"pmids\": [\"32075754\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Ctf4-mediated Pol alpha recruitment for lagging-strand initiation and for origin firing are both sensitive to Pol alpha abundance, but lagging-strand initiation is specifically more dependent on Ctf4-Pol alpha interaction when Pol alpha is limiting, suggesting Pol alpha associates distributively at the fork.\",\n      \"method\": \"Yeast genetics with titratable Pol alpha levels, Okazaki fragment analysis, origin firing efficiency measurement, Ctf4-Pol alpha interaction mutants\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo genetic titration with Okazaki fragment sequencing in a single study\",\n      \"pmids\": [\"32379761\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"WDHD1 (AND-1) acts as an E3 ubiquitin ligase that promotes ubiquitination of MAPRE2 in the nucleus; WDHD1 and MAPRE2 interact by co-immunoprecipitation, and WDHD1 overexpression leads to MAPRE2 degradation conferring cisplatin resistance in lung adenocarcinoma cells.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, WDHD1 knockout, protein stability assay, xenograft\",\n      \"journal\": \"Frontiers in oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP and ubiquitination assay in a single lab; E3 ligase activity not biochemically reconstituted with purified proteins\",\n      \"pmids\": [\"32426268\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"STAT3 transcription factor binds the promoter/upstream regulatory region of WDHD1 gene and activates its transcription; WDHD1 mediates the DNA replication function of STAT3 (STAT3 knockdown reduces DNA replication, which is rescued by WDHD1 expression).\",\n      \"method\": \"ChIP (STAT3 at WDHD1 promoter), siRNA knockdown, ectopic expression, DNA replication assay\",\n      \"journal\": \"Cell & bioscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP plus epistasis rescue experiment in a single lab\",\n      \"pmids\": [\"33413624\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Two AND-1 inhibitors (bazedoxifene acetate and compound CH3) promote AND-1 degradation by disrupting AND-1 polymerization through direct interaction with the WD40 domain; depolymerization promotes interaction with E3 ligase CUL4B, leading to AND-1 ubiquitination and degradation.\",\n      \"method\": \"High-throughput screen, in vitro binding assay (WD40 domain interaction), co-immunoprecipitation (CUL4B interaction), ubiquitination assay, in vitro and in vivo cancer growth assays\",\n      \"journal\": \"Clinical and translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding to WD40 domain and Co-IP for CUL4B interaction established, ubiquitination shown; reconstitution not fully purified\",\n      \"pmids\": [\"34923765\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"AND-1 (WDHD1) is phosphorylated at T826 by ATR in response to interstrand crosslinks (ICLs); this phosphorylation triggers an intramolecular change that promotes AND-1 interaction with FANCM/FAAP24 complex, recruiting it to ICL-stalled forks to activate Fanconi anemia signaling.\",\n      \"method\": \"Co-immunoprecipitation, phosphorylation site mutagenesis (T826A), ATR inhibition, immunofluorescence at ICL foci, proximity ligation assay\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — phospho-site mutagenesis with Co-IP, PLA, and functional FA pathway assays; extends the ATR-phosphorylation mechanism established in the 2015 EMBO J paper\",\n      \"pmids\": [\"35867033\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ZBTB16 transcription factor inhibits WDHD1 transcription; ectopic WDHD1 expression reverses ZBTB16-mediated inhibition of lung adenocarcinoma cell proliferation and S-phase cell cycle arrest, placing WDHD1 downstream of ZBTB16 in a regulatory axis.\",\n      \"method\": \"Ectopic expression, siRNA/overexpression rescue, cell cycle analysis, xenograft, bioinformatics/promoter analysis\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis (WDHD1 rescue of ZBTB16 overexpression phenotype) in a single lab\",\n      \"pmids\": [\"38654107\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"WDHD1/AND-1/CTF4 is a homotrimeric replisome hub protein (trimerization driven by its SepB/beta-propeller domain) that bridges the CMG (Cdc45-MCM-GINS) helicase to DNA polymerase alpha-primase via CIP-box interactions on its helical extensions, thereby coupling DNA unwinding to lagging-strand priming; it is loaded onto chromatin in a Mcm10- and RecQL4-dependent manner after the G1/S transition, stimulates the activities of replicative polymerases in vitro, facilitates parental histone H3-H4 transfer to lagging strands, recruits multiple CIP-box partners (including Chl1 helicase, Dna2, and Sen1) to coordinate sister chromatid cohesion establishment, rDNA copy-number maintenance, and transcription-replication conflict resolution; in response to replication stress, AND-1 is phosphorylated by ATR at T826, enabling it to accumulate at stalled forks and promote Claspin-Chk1 checkpoint activation and FANCM/FAAP24-dependent Fanconi anemia signaling; its WD40 domain specifically protects replication forks from MRE11-dependent resection and DSB formation, while its HMG box confers DNA-binding activity and mediates the unique mammalian interaction with the Pol alpha B subunit.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"WDHD1 (AND-1/CTF4) is a replisome hub protein that physically couples the CMG (Cdc45-MCM2-7-GINS) helicase to DNA polymerase alpha-primase, thereby linking parental-strand unwinding to lagging-strand priming and coordinating downstream genome-maintenance events [#7, #16, #17]. It self-associates into a disk-shaped homotrimer through a C-terminal beta-propeller/SepB domain, and the helical extensions of each protomer dock conserved Ctf4-interacting peptides (CIP-boxes) shared by Pol alpha and the GINS subunit Sld5, allowing one trimer to bridge multiple replisome components [#17, #26]; cryo-EM of the human CMG-AND-1 (CMGA) assembly shows the trimer docking onto Cdc45/GINS via one propeller while its CIP-binding helices remain free for partner recruitment [#36], and the trimer can dimerize two CMG helicases around a central Pol alpha-primase to form a replication factory [#34]. In human cells AND-1 also engages Pol alpha through its HMG box, which binds the Pol alpha B subunit at the SV40 T-antigen site and confers DNA-binding activity that positions Pol alpha on the lagging-strand template [#25]. Loaded onto chromatin after the G1/S transition in an Mcm10- and RecQL4-dependent, CDK/DDK-controlled manner, AND-1 is required for stable CMG assembly and stimulates replicative polymerase activity in vitro [#6, #9, #19, #11]. As a CIP-box hub it recruits the Chl1 helicase, Dna2, and the transcription-termination helicase Sen1 to the fork to support sister chromatid cohesion establishment, rDNA copy-number control, and transcription-replication conflict resolution [#22, #23, #37, #3], and through the Mcm2-Ctf4-Pol alpha axis it directs transfer of parental H3-H4 to lagging strands [#32]. Under replication stress, ATR phosphorylates AND-1 at T826, driving its accumulation at stalled forks to promote Claspin-Chk1 checkpoint activation and, at interstrand crosslinks, FANCM/FAAP24-dependent Fanconi anemia signaling [#20, #42]; its WD40 domain specifically protects forks from MRE11-dependent resection and DSB formation [#33].\",\n  \"teleology\": [\n    {\n      \"year\": 1992,\n      \"claim\": \"Established the founding link between this protein family and the replication machinery by showing the yeast ortholog associates with Pol alpha and is needed for faithful chromosome transmission.\",\n      \"evidence\": \"Yeast genetics and in vitro Pol alpha association with CTF4/CHL15\",\n      \"pmids\": [\"1341195\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural basis for the Pol alpha interaction\", \"Mechanism connecting Pol alpha binding to chromosome loss unresolved\"]\n    },\n    {\n      \"year\": 1997,\n      \"claim\": \"Defined the human protein's domain architecture and biochemical properties, identifying an N-terminal WD-repeat domain, a C-terminal HMG box, four-way-junction DNA binding, and cell-cycle-regulated nuclear localization.\",\n      \"evidence\": \"cDNA cloning, DNA affinity chromatography, EMSA, and immunolocalization\",\n      \"pmids\": [\"9175701\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Replisome role not yet established\", \"Oligomeric state mischaracterized as dimer\", \"No functional assignment to domains\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Genetically positioned the protein in the sister chromatid cohesion establishment pathway acting at the replication fork in both mitosis and meiosis.\",\n      \"evidence\": \"Yeast cohesion and checkpoint-epistasis assays across mitosis and meiosis\",\n      \"pmids\": [\"11287619\", \"12455694\", \"15226378\", \"10101169\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism coupling fork passage to cohesion unknown\", \"Did not identify the direct effectors recruited for cohesion\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Demonstrated the central coupling function: the protein bridges the MCM/GINS helicase to Pol alpha within the replisome, and is required for CMG assembly and replisome stability in human cells.\",\n      \"evidence\": \"Yeast ChIP and in vitro GINS-Ctf4-Pol alpha reconstitution; human BiFC with siRNA and kinase inhibition; Xenopus extract loading after Mcm10\",\n      \"pmids\": [\"19661920\", \"19496828\", \"19805216\", \"17761813\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Atomic interaction interfaces not yet defined\", \"Stoichiometry of the coupling not determined\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Reconstituted the biochemical activity, showing the purified protein binds template-primer DNA, directly contacts Pol alpha/delta/epsilon, and stimulates Pol alpha and epsilon activity.\",\n      \"evidence\": \"Purified-protein DNA-binding and polymerase-stimulation assays plus DNA fiber analysis\",\n      \"pmids\": [\"20089864\", \"20237432\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Oligomeric state reported as dimer, later revised to trimer\", \"How polymerase stimulation operates at the fork in vivo unclear\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Solved the architecture as a homotrimeric hub: a SepB beta-propeller drives trimerization and helical extensions dock shared CIP-box peptides from Pol alpha and GINS, enabling simultaneous binding of multiple replisome partners.\",\n      \"evidence\": \"X-ray crystallography, EM, native mass spectrometry, and mutagenesis in yeast; human SepB/WD40 crystal structures\",\n      \"pmids\": [\"24805245\", \"24255107\", \"28381552\", \"26082189\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo consequences of trimer disruption not fully mapped\", \"Human CMG recruitment route differed from yeast and remained undefined\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Generalized the hub model by showing the trimer recruits additional CIP-box partners (Chl1, Dna2, Tof2, Sen1) to assign distinct genome-maintenance outputs including cohesion, rDNA copy-number control, and conflict resolution.\",\n      \"evidence\": \"Crystallography, EM of reconstituted assemblies, and separation-of-function yeast genetics\",\n      \"pmids\": [\"27397686\", \"27397685\", \"32075754\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Competition/hierarchy among CIP-box partners at a single fork unresolved\", \"Human counterparts of several yeast partner interactions not tested\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Defined the mammalian-specific Pol alpha contact through the HMG box binding the Pol alpha B subunit at the SV40 T-antigen site, distinguishing human AND-1 mechanism from yeast.\",\n      \"evidence\": \"Biochemical interaction assays, domain mutagenesis, and T-antigen peptide competition\",\n      \"pmids\": [\"29167311\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How HMG-box and SepB contacts are coordinated on one trimer unclear\", \"Physiological requirement for the B-subunit contact in cells untested here\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Connected the protein to the DNA-damage response, showing ATR phosphorylation at T826 drives accumulation at stalled forks to promote Claspin-Chk1 activation and that it regulates CtIP-mediated end resection and HR repair.\",\n      \"evidence\": \"Phospho-site mutagenesis, in vitro ssDNA binding, Co-IP, immunofluorescence at damage foci, and HR/resection assays\",\n      \"pmids\": [\"26082189\", \"27940552\", \"27940557\", \"28525744\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How a replisome coupler is repurposed for resection mechanistically incomplete\", \"Relationship between fork-protective and resection-promoting roles unresolved\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Assigned domain-specific genome-protection functions: the Mcm2-Ctf4-Pol alpha axis directs parental H3-H4 transfer to lagging strands, and the WD40 domain protects forks from MRE11-dependent resection and DSB conversion.\",\n      \"evidence\": \"SCAR-seq with separation-of-function mutants; auxin-degron depletion with DNA fiber analysis, EM of fork intermediates, and MRE11 inhibition\",\n      \"pmids\": [\"30244834\", \"30082684\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which WD40 shields forks from MRE11 unknown\", \"Coordination of histone-transfer and fork-protection roles undefined\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Refined the assembly into a replication-factory model and resolved the human CMG docking interface, establishing trimer-mediated dimerization of two CMGs around one Pol alpha-primase.\",\n      \"evidence\": \"Cryo-EM of yeast Ctf4-CMG and 3.3 A human CMGA structures with in vitro reconstitution\",\n      \"pmids\": [\"31589141\", \"32453425\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional importance of CMG dimerization in vivo not established\", \"Identity of human-specific CMG recruitment factors still open\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Extended the ATR-T826 phosphorylation mechanism to interstrand crosslinks, showing it triggers an intramolecular change that recruits FANCM/FAAP24 to activate Fanconi anemia signaling.\",\n      \"evidence\": \"T826A mutagenesis, ATR inhibition, Co-IP, proximity ligation, and FA pathway assays\",\n      \"pmids\": [\"35867033\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural nature of the intramolecular conformational change undefined\", \"Whether the same switch governs both Claspin and FANCM recruitment unresolved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Placed WDHD1 within transcriptional regulatory and cancer contexts, identifying STAT3 and ZBTB16 as upstream regulators and proposing E3-ligase and CUL4B-coupled degradation behaviors.\",\n      \"evidence\": \"ChIP, ectopic expression/rescue, ubiquitination assays, and xenografts in lung adenocarcinoma models\",\n      \"pmids\": [\"33413624\", \"38654107\", \"32426268\", \"34923765\", \"27099318\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Claimed E3 ligase activity not reconstituted with purified components\", \"Mechanistic link between transcriptional regulation and replisome function unclear\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how the identity and ordering of human CMG recruitment factors, the competition among CIP-box partners at individual forks, and the coordination between WDHD1's replicative coupling and its damage-response/fork-protection roles are integrated in vivo.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Human CMG recruitment route distinct from yeast GINS not molecularly identified\", \"Hierarchy of competing CIP-box partners at a single fork unknown\", \"Switch between replicative and repair functions not mechanistically defined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [0, 11, 20, 25, 26]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [7, 16, 17, 23, 34]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [11]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [17, 34, 36]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 9]},\n      {\"term_id\": \"GO:0000228\", \"supporting_discovery_ids\": [13, 16, 19]},\n      {\"term_id\": \"GO:0005654\", \"supporting_discovery_ids\": [10, 33]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-69306\", \"supporting_discovery_ids\": [6, 7, 11, 16, 17]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [20, 27, 28, 33, 42]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [3, 9, 15]},\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [13, 14, 32]}\n    ],\n    \"complexes\": [\n      \"CMG (Cdc45-MCM2-7-GINS) helicase\",\n      \"replisome progression complex\",\n      \"Chl1-Ctf4-GINS assembly\"\n    ],\n    \"partners\": [\n      \"POLA1\",\n      \"GINS (Sld5)\",\n      \"MCM10\",\n      \"RECQL4\",\n      \"DNA2\",\n      \"CHL1\",\n      \"CLSPN\",\n      \"FANCM\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}