{"gene":"REC114","run_date":"2026-06-10T06:43:36","timeline":{"discoveries":[{"year":1993,"finding":"REC114 is a meiosis-specific gene in S. cerevisiae transcribed only during meiosis, with expression dependent on the IME1 gene product, and contains a URS1 consensus sequence in its promoter; a null rec114 mutation abolishes meiotic recombination initiation with no detectable mitotic function.","method":"Northern blot hybridization, genetic null mutation analysis","journal":"Current genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct genetic and transcriptional analysis, single lab, two orthogonal methods (Northern blot + genetic complementation)","pmids":["8385581"],"is_preprint":false},{"year":1997,"finding":"REC114 contains a functional intron at its 3' end with a non-consensus AAG splice site; the intron is not essential for expression or meiotic function (intronless copy complements null mutation), but is evolutionarily conserved.","method":"Minigene/intron analysis, genetic complementation","journal":"Molecular & general genetics : MGG","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct functional complementation with intronless construct, single lab","pmids":["9267437"],"is_preprint":false},{"year":1999,"finding":"High copy numbers of REC114 suppress meiotic DSB formation in budding yeast, indicating that over-expression of Rec114 prevents formation of DSB recombination intermediates.","method":"High-copy suppressor screen, genetic analysis of DSB intermediates","journal":"Genes to cells : devoted to molecular & cellular mechanisms","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with functional DSB readout, single lab","pmids":["10526232"],"is_preprint":false},{"year":2006,"finding":"Mer2, Mei4, and Rec114 form a distinct protein complex required for meiotic DSB formation in S. cerevisiae; all three proteins co-immunoprecipitate, localize to overlapping foci on meiotic chromosomes, and are distinct from the Spo11-Ski8 and Mre11-Rad50-Xrs2 sub-complexes.","method":"Co-immunoprecipitation, chromosome immunofluorescence/co-localization","journal":"Genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP plus cytological co-localization, replicated across multiple labs","pmids":["16783010"],"is_preprint":false},{"year":2007,"finding":"Rec114 localizes to meiotic chromosomes (leptonema through early pachynema) independently of other DSB factors, preferentially associating with chromatin loops rather than the cohesin axis; Mei4 localization is strongly dependent on Rec114 and Mer2.","method":"Chromosome immunofluorescence, co-localization with Rec8 (cohesin), systematic deletion analysis","journal":"Chromosoma","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct localization experiments with genetic dependency testing, single lab","pmids":["17558514"],"is_preprint":false},{"year":2007,"finding":"Rec102, Rec104, and Rec114 are required for Spo11 self-interaction (homodimerization) in vivo during meiosis, as assessed by chromatin-bound complexes at DSB sites; Rec102 interacts with Rec114 and Mei4, suggesting Rec102-Rec104 connect the Mei4-Rec114 subgroup to Spo11.","method":"Co-immunoprecipitation of differentially tagged Spo11 proteins, two-hybrid interaction analysis, genetic dependence assays","journal":"Nucleic acids research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP with tagged constructs, single lab, multiple approaches","pmids":["17264124"],"is_preprint":false},{"year":2013,"finding":"Tel1 (ATM) and Mec1 (ATR) phosphorylate Rec114 following Spo11-induced DSB formation; phosphomimetic rec114 mutations reduce Rec114 interaction with DSB hotspot DNA and reduce/delay DSB formation, while non-phosphorylatable rec114 alleles cause genome-wide increases in both DSB levels and Rec114-hotspot association, establishing a feedback mechanism for DSB homeostasis.","method":"Phosphorylation site mutagenesis, ChIP (chromatin immunoprecipitation) at DSB hotspots, DSB quantification, genetic analysis in tel1/mec1 mutants","journal":"PLoS genetics","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — direct phosphosite mutagenesis with multiple orthogonal readouts (ChIP, DSB quantification, genetic epistasis), single lab but comprehensive","pmids":["23825959"],"is_preprint":false},{"year":2013,"finding":"Ndt80, a meiosis-specific transcription factor, contributes to Rec114 degradation late in meiosis, providing a third independent mechanism for down-regulating Rec114 activity.","method":"Genetic analysis in ndt80 mutants, protein level assays","journal":"PLoS genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with protein level readout, single lab","pmids":["23825959"],"is_preprint":false},{"year":2018,"finding":"Mouse REC114 is essential for meiotic DSB formation; MEI4 forms a stable complex with REC114 and IHO1 in mouse spermatocytes; the REC114 C-terminal domain directly binds the MEI4 N-terminal domain in vitro; the REC114 N-terminal domain has structural similarity to Pleckstrin Homology (PH) domains.","method":"Knockout mouse analysis (cytological assessment of DSBs), Co-immunoprecipitation, in vitro protein complex reconstitution, X-ray crystallography/structural determination","journal":"Life science alliance","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution of direct protein-protein interaction, structural determination, KO phenotype, and Co-IP in spermatocytes, single lab with multiple orthogonal methods","pmids":["30569039"],"is_preprint":false},{"year":2019,"finding":"ANKRD31 directly binds REC114 via its PH domain (crystal structure defined); ANKRD31 stabilizes REC114 association with the pseudoautosomal region (PAR) and other genomic locations; loss of ANKRD31 alters DSB locations (especially fails to target DSBs to PAR), delays DSB timing, and dysregulates DSB numbers.","method":"Crystal structure of REC114 PH domain with ANKRD31, knockout mouse analysis, cytological DSB localization and quantification","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure defining intermolecular contacts, KO mouse with multiple mechanistic readouts, single lab","pmids":["31003867"],"is_preprint":false},{"year":2019,"finding":"A human REC114 missense mutation (p.C133G) reduces protein levels in vitro and causes loss of its function to protect its partner MEI4 from degradation; a splice-site mutation (c.546+5G>A) disrupts normal alternative splicing of REC114. Both result in multiple pronuclei formation and early embryonic arrest.","method":"Immunoprecipitation, western blotting, minigene splicing assay in HEK293T cells","journal":"Journal of medical genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus western blot plus splicing assay, single lab, multiple orthogonal methods","pmids":["31704776"],"is_preprint":false},{"year":2022,"finding":"REC114 is a direct binding partner of TOPOVIBL (the SPO11 partner in the mammalian TOPOVIL catalytic complex); structural analysis identified conserved interacting domains between REC114 and TOPOVIBL; point mutations in TOPOVIBL that disrupt REC114 binding strongly reduce DSB activity genome-wide in oocytes and in sub-telomeric regions in spermatocytes, establishing REC114 as a key regulatory component of the TOPOVIL catalytic complex.","method":"Structural analysis of interacting domains, point mutant mouse models, genome-wide DSB mapping","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1 / Strong — structural domain identification, in vivo point-mutant validation with genome-wide DSB readouts, multiple genetic configurations tested","pmids":["36396648"],"is_preprint":false},{"year":2023,"finding":"Mouse REC114 forms homodimers and associates with MEI4 as a 2:1 REC114:MEI4 heterotrimer that further dimerizes; IHO1 forms coiled-coil-based tetramers and directly interacts with the PH domain of REC114 at the same surface recognized by TOPOVIBL and ANKRD31, suggesting REC114 is a regulatory platform mediating mutually exclusive interactions.","method":"AlphaFold2 modeling, biochemical characterization, size-exclusion chromatography, Co-immunoprecipitation","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — AlphaFold2 models validated by biochemical assays, direct binding experiments establishing competition at REC114 PH domain surface","pmids":["37431931"],"is_preprint":false},{"year":2023,"finding":"The Rec114 C-terminus forms alpha-helical dimers that cup the Mei4 N-terminal alpha-helix in a heterotrimeric complex; Rec114-Mei4 contains two DNA-binding sites pointing in opposite directions that drive DNA condensation; Rec114-Mei4 preferentially binds branched DNA substrates; these structural features are conserved across eukaryotes.","method":"AlphaFold2 modeling, NMR spectroscopy, SAXS, mutagenesis, DNA binding assays","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 1 / Strong — NMR, SAXS, and mutagenesis together with functional DNA binding assays, multiple orthogonal structural and biochemical methods","pmids":["37442581"],"is_preprint":false},{"year":2023,"finding":"The minimal Rec114-Mei4 heterotrimeric complex (lacking Rec114 intrinsically disordered region) is sufficient to bind DNA and form condensates; single-molecule experiments show the complex bridges two or more DNA duplexes and generates force to condense DNA through long-range interactions.","method":"In vitro reconstitution of minimal complex, DNA binding assays, condensate formation assays, single-molecule force experiments","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with single-molecule mechanistic experiments, directly probing DNA bridging activity","pmids":["37442580"],"is_preprint":false},{"year":2023,"finding":"The ANKRD31-REC114 interaction is essential for ANKRD31 function in meiosis; complete disruption of this interaction mimics Ankrd31 null phenotype (delayed DSBs, altered DSB locations, PAR targeting failure); ANKRD31 functions as a scaffold that requires REC114 binding for its activity, with a dosage-sensitive relationship.","method":"Targeted Ankrd31 missense and truncation mouse mutants, cytological DSB analysis, crossover mapping","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple engineered mouse alleles with graded disruption of specific protein-protein interaction, multiple orthogonal phenotypic readouts","pmids":["37976262"],"is_preprint":false},{"year":2023,"finding":"A truncating human REC114 variant (p.Gln190*) causes meiotic arrest and non-obstructive azoospermia; the truncated REC114 protein has impaired interaction with MEI4, establishing that the REC114-MEI4 complex is indispensable for meiotic DSB homeostasis in human males.","method":"Co-immunoprecipitation, western blot, testicular histopathology, meiotic chromosome spread analysis","journal":"Clinical genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP demonstrating disrupted protein interaction combined with histological meiotic arrest phenotype, single lab","pmids":["38148155"],"is_preprint":false}],"current_model":"REC114 is an evolutionarily conserved, meiosis-specific protein that forms a 2:1 heterotrimeric complex with MEI4 (stabilizing MEI4 from degradation) and associates with IHO1/Mer2 on meiotic chromosome axes, where its PH-like N-terminal domain serves as a regulatory platform for mutually exclusive interactions with TOPOVIBL (the SPO11 catalytic partner), ANKRD31, and IHO1; the Rec114-MEI4 complex directly binds and bridges DNA duplexes to form condensates that promote assembly of the meiotic DSB machinery, while DSB levels are feedback-regulated through ATM/ATR (Tel1/Mec1) phosphorylation of REC114 that reduces its association with DSB hotspot chromatin, and through Ndt80-dependent REC114 degradation, collectively maintaining DSB homeostasis essential for fertility."},"narrative":{"mechanistic_narrative":"REC114 is an evolutionarily conserved, meiosis-specific protein that nucleates assembly of the machinery generating programmed double-strand breaks (DSBs) that initiate meiotic recombination [PMID:8385581, PMID:16783010]. It forms a distinct chromosome-associated complex with MEI4 and Mer2/IHO1 that localizes to meiotic chromatin loops and is required for DSB formation [PMID:16783010, PMID:17558514]; its C-terminal alpha-helical domain dimerizes and directly binds the MEI4 N-terminal helix to build a 2:1 REC114:MEI4 heterotrimer that further dimerizes, with REC114 stabilizing MEI4 against degradation [PMID:30569039, PMID:37431931, PMID:37442581]. The REC114-MEI4 complex carries two oppositely oriented DNA-binding sites that preferentially engage branched DNA, bridge and condense multiple DNA duplexes, and form condensates that promote DSB-machinery assembly [PMID:37442581, PMID:37442580]. The REC114 N-terminal PH-like domain acts as a regulatory platform whose single binding surface mediates mutually exclusive interactions with IHO1, ANKRD31, and TOPOVIBL (the SPO11 catalytic partner) [PMID:30569039, PMID:31003867, PMID:37431931]; ANKRD31 binding controls DSB timing and targeting, including DSBs to the pseudoautosomal region, while TOPOVIBL binding couples REC114 to the SPO11/TOPOVIL catalytic complex to license break formation [PMID:31003867, PMID:36396648, PMID:37431931]. DSB levels are held in homeostasis by negative feedback: Tel1/Mec1 (ATM/ATR) phosphorylation of REC114 reduces its association with DSB hotspot chromatin, and Ndt80 drives REC114 degradation late in meiosis [PMID:23825959]. In humans, truncating and missense REC114 variants that impair MEI4 binding cause meiotic arrest and non-obstructive azoospermia, and additional REC114 mutations are associated with embryonic arrest, establishing the REC114-MEI4 complex as indispensable for human fertility [PMID:31704776, PMID:38148155].","teleology":[{"year":1993,"claim":"Established REC114 as a meiosis-specific factor whose loss abolishes recombination initiation, defining it as a dedicated meiotic gene rather than a general recombination component.","evidence":"Northern blot of meiotic transcription and genetic null analysis in S. cerevisiae","pmids":["8385581"],"confidence":"Medium","gaps":["No molecular function for the protein identified","Mechanism of action in DSB initiation unknown"]},{"year":1999,"claim":"Showed that REC114 dosage is rate-limiting, since over-expression suppresses DSB formation, hinting at a regulatory role in break control.","evidence":"High-copy suppressor screen with DSB intermediate readout in budding yeast","pmids":["10526232"],"confidence":"Medium","gaps":["Molecular basis of dosage sensitivity unresolved","No biochemical partners identified yet"]},{"year":2006,"claim":"Defined REC114 as part of a discrete Mer2-Mei4-Rec114 complex distinct from the Spo11 nuclease module, placing it in a dedicated pre-DSB sub-assembly.","evidence":"Reciprocal Co-IP and chromosome co-localization in S. cerevisiae","pmids":["16783010"],"confidence":"High","gaps":["Stoichiometry and direct vs indirect interactions not resolved","Connection to the Spo11 catalytic complex unclear"]},{"year":2007,"claim":"Localized REC114 to chromatin loops independently of other DSB factors and showed Mei4 chromosome loading depends on it, ordering the assembly pathway.","evidence":"Chromosome immunofluorescence with cohesin co-staining and deletion-dependency tests","pmids":["17558514"],"confidence":"Medium","gaps":["Direct DNA/chromatin binding by REC114 not demonstrated","Mechanism of loop targeting unknown"]},{"year":2007,"claim":"Connected the Mei4-Rec114 subgroup to Spo11 self-interaction via Rec102-Rec104, linking the regulatory module to nuclease activation.","evidence":"Co-IP of tagged Spo11 and two-hybrid analysis with genetic dependence assays","pmids":["17264124"],"confidence":"Medium","gaps":["Direct REC114-Spo11 contact not established","Bridging architecture inferred rather than reconstituted"]},{"year":2013,"claim":"Revealed a DSB homeostasis feedback loop in which Tel1/Mec1 phosphorylation of REC114 reduces hotspot association and Ndt80 drives its degradation, explaining how break levels are bounded.","evidence":"Phosphosite mutagenesis, hotspot ChIP, DSB quantification and ndt80 genetics in yeast","pmids":["23825959"],"confidence":"High","gaps":["Phosphorylation effect on complex structure not defined","Degradation pathway/E3 ligase not identified"]},{"year":2018,"claim":"Demonstrated conservation in mammals, with mouse REC114 essential for DSBs, defined the direct C-terminal REC114–MEI4 interaction, and identified the N-terminal PH-like fold.","evidence":"Knockout mouse DSB cytology, Co-IP in spermatocytes, in vitro reconstitution and crystallography","pmids":["30569039"],"confidence":"High","gaps":["Function of the PH-like domain not yet assigned","How the complex engages DNA undefined"]},{"year":2019,"claim":"Identified the REC114 PH domain as the docking site for ANKRD31, which directs DSB timing and pseudoautosomal-region targeting, giving the domain a defined regulatory function.","evidence":"Crystal structure of REC114 PH domain with ANKRD31 and knockout-mouse DSB mapping","pmids":["31003867"],"confidence":"High","gaps":["Whether PH domain binds other partners at the same surface not yet known","Mechanism of PAR-specific targeting incomplete"]},{"year":2019,"claim":"Linked REC114 directly to human reproductive disease by showing variants that destabilize REC114 or fail to protect MEI4 cause embryonic arrest.","evidence":"IP, western blot and minigene splicing assays in HEK293T cells with patient variants","pmids":["31704776"],"confidence":"Medium","gaps":["In vivo meiotic phenotype of variants not assessed","Causality at organismal level inferred from cell-based assays"]},{"year":2022,"claim":"Established REC114 as a direct regulatory partner of TOPOVIBL within the SPO11/TOPOVIL catalytic complex, coupling the assembly platform to the nuclease.","evidence":"Structural domain mapping and TOPOVIBL point-mutant mice with genome-wide DSB mapping","pmids":["36396648"],"confidence":"High","gaps":["How TOPOVIBL binding triggers catalysis not resolved","Temporal order relative to ANKRD31/IHO1 binding unclear"]},{"year":2023,"claim":"Resolved the heterotrimer architecture and showed the REC114 PH-domain surface mediates mutually exclusive binding to IHO1, ANKRD31 and TOPOVIBL, recasting REC114 as a competitive regulatory hub.","evidence":"AlphaFold2 modeling validated by SEC, biochemistry and Co-IP","pmids":["37431931"],"confidence":"High","gaps":["What controls partner exchange in vivo unknown","Temporal regulation of competing interactions not defined"]},{"year":2023,"claim":"Defined the structural basis of REC114-MEI4 DNA engagement, showing two oppositely oriented DNA-binding sites and preference for branched DNA that drive condensation.","evidence":"AlphaFold2, NMR, SAXS, mutagenesis and DNA-binding assays","pmids":["37442581"],"confidence":"High","gaps":["In vivo relevance of branched-DNA preference not tested","Link between condensation and Spo11 activation incomplete"]},{"year":2023,"claim":"Demonstrated that a minimal REC114-MEI4 complex bridges and condenses DNA duplexes via long-range force, providing a biophysical mechanism for organizing DSB machinery.","evidence":"In vitro reconstitution with condensate and single-molecule force experiments","pmids":["37442580"],"confidence":"High","gaps":["Physiological condensate composition not defined","How bridging promotes Spo11 cleavage unresolved"]},{"year":2023,"claim":"Showed genetically that the ANKRD31-REC114 interaction is required and dosage-sensitive for ANKRD31 scaffold function, validating the structural interface in vivo.","evidence":"Engineered Ankrd31 missense/truncation mice with DSB and crossover analysis","pmids":["37976262"],"confidence":"High","gaps":["Stoichiometric basis of dosage sensitivity unresolved","Whether REC114 limits or enables ANKRD31 at distinct loci unclear"]},{"year":2023,"claim":"Confirmed in human males that a truncating REC114 variant disrupting MEI4 binding causes meiotic arrest and azoospermia, cementing the complex's role in human fertility.","evidence":"Co-IP, western blot, testicular histopathology and meiotic spreads from patients","pmids":["38148155"],"confidence":"Medium","gaps":["Single-family genetic evidence","Quantitative effect on DSB numbers in patients not measured"]},{"year":null,"claim":"How the competing PH-domain interactions, phosphorylation feedback, and condensate formation are temporally coordinated to license a precise number and distribution of DSBs remains unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No integrated in vivo timeline of partner exchange","Mechanistic link between condensation and Spo11 catalysis missing","Regulatory inputs governing DSB number not fully mapped"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[13,14]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[8,9,12]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[11,12]}],"localization":[{"term_id":"GO:0005694","term_label":"chromosome","supporting_discovery_ids":[3,4]},{"term_id":"GO:0000228","term_label":"nuclear chromosome","supporting_discovery_ids":[4]}],"pathway":[{"term_id":"R-HSA-1474165","term_label":"Reproduction","supporting_discovery_ids":[8,16]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,3]}],"complexes":["REC114-MEI4 heterotrimer","Mer2/IHO1-MEI4-REC114 pre-DSB complex","SPO11/TOPOVIL catalytic complex"],"partners":["MEI4","IHO1","MER2","ANKRD31","TOPOVIBL","REC102","SPO11"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q7Z4M0","full_name":"Meiotic recombination protein REC114","aliases":[],"length_aa":266,"mass_kda":29.2,"function":"Required for DNA double-strand breaks (DSBs) formation in unsynapsed regions during meiotic recombination (PubMed:38148155). Probably acts by forming a complex with IHO1 and MEI4, which activates DSBs formation in unsynapsed regions, an essential step to ensure completion of synapsis (By similarity). Required for spermatogenesis (PubMed:38148155). Required for oogenesis","subcellular_location":"Chromosome","url":"https://www.uniprot.org/uniprotkb/Q7Z4M0/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/REC114","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/REC114","total_profiled":1310},"omim":[{"mim_id":"619190","title":"INTERACTOR OF HORMAD1 1; IHO1","url":"https://www.omim.org/entry/619190"},{"mim_id":"619176","title":"OOCYTE/ZYGOTE/EMBRYO MATURATION ARREST 10; OZEMA10","url":"https://www.omim.org/entry/619176"},{"mim_id":"618423","title":"ANKYRIN REPEAT DOMAIN-CONTAINING PROTEIN 31; ANKRD31","url":"https://www.omim.org/entry/618423"},{"mim_id":"618421","title":"REC114 MEIOTIC RECOMBINATION PROTEIN; REC114","url":"https://www.omim.org/entry/618421"},{"mim_id":"618417","title":"MEIOTIC DOUBLE-STRANDED BREAK FORMATION PROTEIN 4; MEI4","url":"https://www.omim.org/entry/618417"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enriched","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"testis","ntpm":37.5}],"url":"https://www.proteinatlas.org/search/REC114"},"hgnc":{"alias_symbol":["LOC283677","FLJ27520","FLJ36860","FLJ44083","CT147"],"prev_symbol":["C15orf60"]},"alphafold":{"accession":"Q7Z4M0","domains":[{"cath_id":"2.30.29.30","chopping":"19-33_50-148","consensus_level":"high","plddt":93.3244,"start":19,"end":148},{"cath_id":"-","chopping":"206-222_241-266","consensus_level":"medium","plddt":82.7363,"start":206,"end":266}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q7Z4M0","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q7Z4M0-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q7Z4M0-F1-predicted_aligned_error_v6.png","plddt_mean":72.38},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=REC114","jax_strain_url":"https://www.jax.org/strain/search?query=REC114"},"sequence":{"accession":"Q7Z4M0","fasta_url":"https://rest.uniprot.org/uniprotkb/Q7Z4M0.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q7Z4M0/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q7Z4M0"}},"corpus_meta":[{"pmid":"17558514","id":"PMC_17558514","title":"Interactions between Mei4, Rec114, and other proteins required for meiotic DNA double-strand break formation in Saccharomyces cerevisiae.","date":"2007","source":"Chromosoma","url":"https://pubmed.ncbi.nlm.nih.gov/17558514","citation_count":111,"is_preprint":false},{"pmid":"23825959","id":"PMC_23825959","title":"Budding yeast ATM/ATR control meiotic double-strand break (DSB) levels by down-regulating Rec114, an essential component of the DSB-machinery.","date":"2013","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/23825959","citation_count":100,"is_preprint":false},{"pmid":"16783010","id":"PMC_16783010","title":"Saccharomyces cerevisiae Mer2, Mei4 and Rec114 form a complex required for meiotic double-strand break formation.","date":"2006","source":"Genetics","url":"https://pubmed.ncbi.nlm.nih.gov/16783010","citation_count":92,"is_preprint":false},{"pmid":"31003867","id":"PMC_31003867","title":"REC114 Partner ANKRD31 Controls Number, Timing, and Location of Meiotic DNA Breaks.","date":"2019","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/31003867","citation_count":91,"is_preprint":false},{"pmid":"10526232","id":"PMC_10526232","title":"High copy number suppression of the meiotic arrest caused by a dmc1 mutation: REC114 imposes an early recombination block and RAD54 promotes a DMC1-independent DSB repair pathway.","date":"1999","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/10526232","citation_count":81,"is_preprint":false},{"pmid":"30569039","id":"PMC_30569039","title":"Mouse REC114 is essential for meiotic DNA double-strand break formation and forms a complex with MEI4.","date":"2018","source":"Life science alliance","url":"https://pubmed.ncbi.nlm.nih.gov/30569039","citation_count":80,"is_preprint":false},{"pmid":"31704776","id":"PMC_31704776","title":"Homozygous mutations in REC114 cause female infertility characterised by multiple pronuclei formation and early embryonic arrest.","date":"2019","source":"Journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/31704776","citation_count":55,"is_preprint":false},{"pmid":"17264124","id":"PMC_17264124","title":"Meiotic association between Spo11 regulated by Rec102, Rec104 and Rec114.","date":"2007","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/17264124","citation_count":48,"is_preprint":false},{"pmid":"36396648","id":"PMC_36396648","title":"TOPOVIBL-REC114 interaction regulates meiotic DNA double-strand breaks.","date":"2022","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/36396648","citation_count":36,"is_preprint":false},{"pmid":"8385581","id":"PMC_8385581","title":"Genetic and molecular analysis of REC114, an early meiotic recombination gene in yeast.","date":"1993","source":"Current genetics","url":"https://pubmed.ncbi.nlm.nih.gov/8385581","citation_count":27,"is_preprint":false},{"pmid":"9267437","id":"PMC_9267437","title":"Examination of the intron in the meiosis-specific recombination gene REC114 in Saccharomyces.","date":"1997","source":"Molecular & general genetics : MGG","url":"https://pubmed.ncbi.nlm.nih.gov/9267437","citation_count":21,"is_preprint":false},{"pmid":"37442581","id":"PMC_37442581","title":"Evolutionary conservation of the structure and function of meiotic Rec114-Mei4 and Mer2 complexes.","date":"2023","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/37442581","citation_count":19,"is_preprint":false},{"pmid":"37431931","id":"PMC_37431931","title":"Characterization of the REC114-MEI4-IHO1 complex regulating meiotic DNA double-strand break formation.","date":"2023","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/37431931","citation_count":18,"is_preprint":false},{"pmid":"37976262","id":"PMC_37976262","title":"Essential roles of the ANKRD31-REC114 interaction in meiotic recombination and mouse spermatogenesis.","date":"2023","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/37976262","citation_count":13,"is_preprint":false},{"pmid":"37442580","id":"PMC_37442580","title":"Structure and DNA-bridging activity of the essential Rec114-Mei4 trimer interface.","date":"2023","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/37442580","citation_count":13,"is_preprint":false},{"pmid":"38148155","id":"PMC_38148155","title":"A bi-allelic REC114 loss-of-function variant causes meiotic arrest and nonobstructive azoospermia.","date":"2023","source":"Clinical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/38148155","citation_count":7,"is_preprint":false},{"pmid":"32290544","id":"PMC_32290544","title":"Genetic Interactions of Histone Modification Machinery Set1 and PAF1C with the Recombination Complex Rec114-Mer2-Mei4 in the Formation of Meiotic DNA Double-Strand Breaks.","date":"2020","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/32290544","citation_count":6,"is_preprint":false},{"pmid":"37162821","id":"PMC_37162821","title":"Essential roles of the ANKRD31-REC114 interaction in meiotic recombination and mouse spermatogenesis.","date":"2023","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/37162821","citation_count":1,"is_preprint":false},{"pmid":"41168699","id":"PMC_41168699","title":"Compound heterozygous REC114 variants in dizygotic twins causes meiotic arrest and non-obstructive azoospermia.","date":"2025","source":"Basic and clinical andrology","url":"https://pubmed.ncbi.nlm.nih.gov/41168699","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.07.08.663703","title":"Recruitment of Mre11 to recombination sites during meiosis","date":"2025-07-08","source":"bioRxiv","url":"https://doi.org/10.1101/2025.07.08.663703","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.06.20.599484","title":"Individual variation in meiotic crossover positioning, rate and interference are associated with distinct genetic processes in domestic pigs","date":"2024-06-24","source":"bioRxiv","url":"https://doi.org/10.1101/2024.06.20.599484","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11971,"output_tokens":3899,"usd":0.047199,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11738,"output_tokens":4378,"usd":0.08407,"stage2_stop_reason":"end_turn"},"total_usd":0.131269,"stage1_batch_id":"msgbatch_01CNoDbkChHEkgrJnL7BwtGJ","stage2_batch_id":"msgbatch_016Y4x9E9Wg5VQDANj9quSQD","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1993,\n      \"finding\": \"REC114 is a meiosis-specific gene in S. cerevisiae transcribed only during meiosis, with expression dependent on the IME1 gene product, and contains a URS1 consensus sequence in its promoter; a null rec114 mutation abolishes meiotic recombination initiation with no detectable mitotic function.\",\n      \"method\": \"Northern blot hybridization, genetic null mutation analysis\",\n      \"journal\": \"Current genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct genetic and transcriptional analysis, single lab, two orthogonal methods (Northern blot + genetic complementation)\",\n      \"pmids\": [\"8385581\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"REC114 contains a functional intron at its 3' end with a non-consensus AAG splice site; the intron is not essential for expression or meiotic function (intronless copy complements null mutation), but is evolutionarily conserved.\",\n      \"method\": \"Minigene/intron analysis, genetic complementation\",\n      \"journal\": \"Molecular & general genetics : MGG\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct functional complementation with intronless construct, single lab\",\n      \"pmids\": [\"9267437\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"High copy numbers of REC114 suppress meiotic DSB formation in budding yeast, indicating that over-expression of Rec114 prevents formation of DSB recombination intermediates.\",\n      \"method\": \"High-copy suppressor screen, genetic analysis of DSB intermediates\",\n      \"journal\": \"Genes to cells : devoted to molecular & cellular mechanisms\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with functional DSB readout, single lab\",\n      \"pmids\": [\"10526232\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Mer2, Mei4, and Rec114 form a distinct protein complex required for meiotic DSB formation in S. cerevisiae; all three proteins co-immunoprecipitate, localize to overlapping foci on meiotic chromosomes, and are distinct from the Spo11-Ski8 and Mre11-Rad50-Xrs2 sub-complexes.\",\n      \"method\": \"Co-immunoprecipitation, chromosome immunofluorescence/co-localization\",\n      \"journal\": \"Genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP plus cytological co-localization, replicated across multiple labs\",\n      \"pmids\": [\"16783010\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Rec114 localizes to meiotic chromosomes (leptonema through early pachynema) independently of other DSB factors, preferentially associating with chromatin loops rather than the cohesin axis; Mei4 localization is strongly dependent on Rec114 and Mer2.\",\n      \"method\": \"Chromosome immunofluorescence, co-localization with Rec8 (cohesin), systematic deletion analysis\",\n      \"journal\": \"Chromosoma\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization experiments with genetic dependency testing, single lab\",\n      \"pmids\": [\"17558514\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Rec102, Rec104, and Rec114 are required for Spo11 self-interaction (homodimerization) in vivo during meiosis, as assessed by chromatin-bound complexes at DSB sites; Rec102 interacts with Rec114 and Mei4, suggesting Rec102-Rec104 connect the Mei4-Rec114 subgroup to Spo11.\",\n      \"method\": \"Co-immunoprecipitation of differentially tagged Spo11 proteins, two-hybrid interaction analysis, genetic dependence assays\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP with tagged constructs, single lab, multiple approaches\",\n      \"pmids\": [\"17264124\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Tel1 (ATM) and Mec1 (ATR) phosphorylate Rec114 following Spo11-induced DSB formation; phosphomimetic rec114 mutations reduce Rec114 interaction with DSB hotspot DNA and reduce/delay DSB formation, while non-phosphorylatable rec114 alleles cause genome-wide increases in both DSB levels and Rec114-hotspot association, establishing a feedback mechanism for DSB homeostasis.\",\n      \"method\": \"Phosphorylation site mutagenesis, ChIP (chromatin immunoprecipitation) at DSB hotspots, DSB quantification, genetic analysis in tel1/mec1 mutants\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — direct phosphosite mutagenesis with multiple orthogonal readouts (ChIP, DSB quantification, genetic epistasis), single lab but comprehensive\",\n      \"pmids\": [\"23825959\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Ndt80, a meiosis-specific transcription factor, contributes to Rec114 degradation late in meiosis, providing a third independent mechanism for down-regulating Rec114 activity.\",\n      \"method\": \"Genetic analysis in ndt80 mutants, protein level assays\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with protein level readout, single lab\",\n      \"pmids\": [\"23825959\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Mouse REC114 is essential for meiotic DSB formation; MEI4 forms a stable complex with REC114 and IHO1 in mouse spermatocytes; the REC114 C-terminal domain directly binds the MEI4 N-terminal domain in vitro; the REC114 N-terminal domain has structural similarity to Pleckstrin Homology (PH) domains.\",\n      \"method\": \"Knockout mouse analysis (cytological assessment of DSBs), Co-immunoprecipitation, in vitro protein complex reconstitution, X-ray crystallography/structural determination\",\n      \"journal\": \"Life science alliance\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution of direct protein-protein interaction, structural determination, KO phenotype, and Co-IP in spermatocytes, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"30569039\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"ANKRD31 directly binds REC114 via its PH domain (crystal structure defined); ANKRD31 stabilizes REC114 association with the pseudoautosomal region (PAR) and other genomic locations; loss of ANKRD31 alters DSB locations (especially fails to target DSBs to PAR), delays DSB timing, and dysregulates DSB numbers.\",\n      \"method\": \"Crystal structure of REC114 PH domain with ANKRD31, knockout mouse analysis, cytological DSB localization and quantification\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure defining intermolecular contacts, KO mouse with multiple mechanistic readouts, single lab\",\n      \"pmids\": [\"31003867\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"A human REC114 missense mutation (p.C133G) reduces protein levels in vitro and causes loss of its function to protect its partner MEI4 from degradation; a splice-site mutation (c.546+5G>A) disrupts normal alternative splicing of REC114. Both result in multiple pronuclei formation and early embryonic arrest.\",\n      \"method\": \"Immunoprecipitation, western blotting, minigene splicing assay in HEK293T cells\",\n      \"journal\": \"Journal of medical genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus western blot plus splicing assay, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"31704776\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"REC114 is a direct binding partner of TOPOVIBL (the SPO11 partner in the mammalian TOPOVIL catalytic complex); structural analysis identified conserved interacting domains between REC114 and TOPOVIBL; point mutations in TOPOVIBL that disrupt REC114 binding strongly reduce DSB activity genome-wide in oocytes and in sub-telomeric regions in spermatocytes, establishing REC114 as a key regulatory component of the TOPOVIL catalytic complex.\",\n      \"method\": \"Structural analysis of interacting domains, point mutant mouse models, genome-wide DSB mapping\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — structural domain identification, in vivo point-mutant validation with genome-wide DSB readouts, multiple genetic configurations tested\",\n      \"pmids\": [\"36396648\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Mouse REC114 forms homodimers and associates with MEI4 as a 2:1 REC114:MEI4 heterotrimer that further dimerizes; IHO1 forms coiled-coil-based tetramers and directly interacts with the PH domain of REC114 at the same surface recognized by TOPOVIBL and ANKRD31, suggesting REC114 is a regulatory platform mediating mutually exclusive interactions.\",\n      \"method\": \"AlphaFold2 modeling, biochemical characterization, size-exclusion chromatography, Co-immunoprecipitation\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — AlphaFold2 models validated by biochemical assays, direct binding experiments establishing competition at REC114 PH domain surface\",\n      \"pmids\": [\"37431931\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The Rec114 C-terminus forms alpha-helical dimers that cup the Mei4 N-terminal alpha-helix in a heterotrimeric complex; Rec114-Mei4 contains two DNA-binding sites pointing in opposite directions that drive DNA condensation; Rec114-Mei4 preferentially binds branched DNA substrates; these structural features are conserved across eukaryotes.\",\n      \"method\": \"AlphaFold2 modeling, NMR spectroscopy, SAXS, mutagenesis, DNA binding assays\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — NMR, SAXS, and mutagenesis together with functional DNA binding assays, multiple orthogonal structural and biochemical methods\",\n      \"pmids\": [\"37442581\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The minimal Rec114-Mei4 heterotrimeric complex (lacking Rec114 intrinsically disordered region) is sufficient to bind DNA and form condensates; single-molecule experiments show the complex bridges two or more DNA duplexes and generates force to condense DNA through long-range interactions.\",\n      \"method\": \"In vitro reconstitution of minimal complex, DNA binding assays, condensate formation assays, single-molecule force experiments\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with single-molecule mechanistic experiments, directly probing DNA bridging activity\",\n      \"pmids\": [\"37442580\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The ANKRD31-REC114 interaction is essential for ANKRD31 function in meiosis; complete disruption of this interaction mimics Ankrd31 null phenotype (delayed DSBs, altered DSB locations, PAR targeting failure); ANKRD31 functions as a scaffold that requires REC114 binding for its activity, with a dosage-sensitive relationship.\",\n      \"method\": \"Targeted Ankrd31 missense and truncation mouse mutants, cytological DSB analysis, crossover mapping\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple engineered mouse alleles with graded disruption of specific protein-protein interaction, multiple orthogonal phenotypic readouts\",\n      \"pmids\": [\"37976262\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"A truncating human REC114 variant (p.Gln190*) causes meiotic arrest and non-obstructive azoospermia; the truncated REC114 protein has impaired interaction with MEI4, establishing that the REC114-MEI4 complex is indispensable for meiotic DSB homeostasis in human males.\",\n      \"method\": \"Co-immunoprecipitation, western blot, testicular histopathology, meiotic chromosome spread analysis\",\n      \"journal\": \"Clinical genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP demonstrating disrupted protein interaction combined with histological meiotic arrest phenotype, single lab\",\n      \"pmids\": [\"38148155\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"REC114 is an evolutionarily conserved, meiosis-specific protein that forms a 2:1 heterotrimeric complex with MEI4 (stabilizing MEI4 from degradation) and associates with IHO1/Mer2 on meiotic chromosome axes, where its PH-like N-terminal domain serves as a regulatory platform for mutually exclusive interactions with TOPOVIBL (the SPO11 catalytic partner), ANKRD31, and IHO1; the Rec114-MEI4 complex directly binds and bridges DNA duplexes to form condensates that promote assembly of the meiotic DSB machinery, while DSB levels are feedback-regulated through ATM/ATR (Tel1/Mec1) phosphorylation of REC114 that reduces its association with DSB hotspot chromatin, and through Ndt80-dependent REC114 degradation, collectively maintaining DSB homeostasis essential for fertility.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"REC114 is an evolutionarily conserved, meiosis-specific protein that nucleates assembly of the machinery generating programmed double-strand breaks (DSBs) that initiate meiotic recombination [#0, #3]. It forms a distinct chromosome-associated complex with MEI4 and Mer2/IHO1 that localizes to meiotic chromatin loops and is required for DSB formation [#3, #4]; its C-terminal alpha-helical domain dimerizes and directly binds the MEI4 N-terminal helix to build a 2:1 REC114:MEI4 heterotrimer that further dimerizes, with REC114 stabilizing MEI4 against degradation [#8, #12, #13]. The REC114-MEI4 complex carries two oppositely oriented DNA-binding sites that preferentially engage branched DNA, bridge and condense multiple DNA duplexes, and form condensates that promote DSB-machinery assembly [#13, #14]. The REC114 N-terminal PH-like domain acts as a regulatory platform whose single binding surface mediates mutually exclusive interactions with IHO1, ANKRD31, and TOPOVIBL (the SPO11 catalytic partner) [#8, #9, #12]; ANKRD31 binding controls DSB timing and targeting, including DSBs to the pseudoautosomal region, while TOPOVIBL binding couples REC114 to the SPO11/TOPOVIL catalytic complex to license break formation [#9, #11, #12]. DSB levels are held in homeostasis by negative feedback: Tel1/Mec1 (ATM/ATR) phosphorylation of REC114 reduces its association with DSB hotspot chromatin, and Ndt80 drives REC114 degradation late in meiosis [#6, #7]. In humans, truncating and missense REC114 variants that impair MEI4 binding cause meiotic arrest and non-obstructive azoospermia, and additional REC114 mutations are associated with embryonic arrest, establishing the REC114-MEI4 complex as indispensable for human fertility [#10, #16].\",\n  \"teleology\": [\n    {\n      \"year\": 1993,\n      \"claim\": \"Established REC114 as a meiosis-specific factor whose loss abolishes recombination initiation, defining it as a dedicated meiotic gene rather than a general recombination component.\",\n      \"evidence\": \"Northern blot of meiotic transcription and genetic null analysis in S. cerevisiae\",\n      \"pmids\": [\"8385581\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No molecular function for the protein identified\", \"Mechanism of action in DSB initiation unknown\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Showed that REC114 dosage is rate-limiting, since over-expression suppresses DSB formation, hinting at a regulatory role in break control.\",\n      \"evidence\": \"High-copy suppressor screen with DSB intermediate readout in budding yeast\",\n      \"pmids\": [\"10526232\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular basis of dosage sensitivity unresolved\", \"No biochemical partners identified yet\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Defined REC114 as part of a discrete Mer2-Mei4-Rec114 complex distinct from the Spo11 nuclease module, placing it in a dedicated pre-DSB sub-assembly.\",\n      \"evidence\": \"Reciprocal Co-IP and chromosome co-localization in S. cerevisiae\",\n      \"pmids\": [\"16783010\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry and direct vs indirect interactions not resolved\", \"Connection to the Spo11 catalytic complex unclear\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Localized REC114 to chromatin loops independently of other DSB factors and showed Mei4 chromosome loading depends on it, ordering the assembly pathway.\",\n      \"evidence\": \"Chromosome immunofluorescence with cohesin co-staining and deletion-dependency tests\",\n      \"pmids\": [\"17558514\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct DNA/chromatin binding by REC114 not demonstrated\", \"Mechanism of loop targeting unknown\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Connected the Mei4-Rec114 subgroup to Spo11 self-interaction via Rec102-Rec104, linking the regulatory module to nuclease activation.\",\n      \"evidence\": \"Co-IP of tagged Spo11 and two-hybrid analysis with genetic dependence assays\",\n      \"pmids\": [\"17264124\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct REC114-Spo11 contact not established\", \"Bridging architecture inferred rather than reconstituted\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Revealed a DSB homeostasis feedback loop in which Tel1/Mec1 phosphorylation of REC114 reduces hotspot association and Ndt80 drives its degradation, explaining how break levels are bounded.\",\n      \"evidence\": \"Phosphosite mutagenesis, hotspot ChIP, DSB quantification and ndt80 genetics in yeast\",\n      \"pmids\": [\"23825959\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Phosphorylation effect on complex structure not defined\", \"Degradation pathway/E3 ligase not identified\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Demonstrated conservation in mammals, with mouse REC114 essential for DSBs, defined the direct C-terminal REC114–MEI4 interaction, and identified the N-terminal PH-like fold.\",\n      \"evidence\": \"Knockout mouse DSB cytology, Co-IP in spermatocytes, in vitro reconstitution and crystallography\",\n      \"pmids\": [\"30569039\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Function of the PH-like domain not yet assigned\", \"How the complex engages DNA undefined\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified the REC114 PH domain as the docking site for ANKRD31, which directs DSB timing and pseudoautosomal-region targeting, giving the domain a defined regulatory function.\",\n      \"evidence\": \"Crystal structure of REC114 PH domain with ANKRD31 and knockout-mouse DSB mapping\",\n      \"pmids\": [\"31003867\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether PH domain binds other partners at the same surface not yet known\", \"Mechanism of PAR-specific targeting incomplete\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Linked REC114 directly to human reproductive disease by showing variants that destabilize REC114 or fail to protect MEI4 cause embryonic arrest.\",\n      \"evidence\": \"IP, western blot and minigene splicing assays in HEK293T cells with patient variants\",\n      \"pmids\": [\"31704776\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"In vivo meiotic phenotype of variants not assessed\", \"Causality at organismal level inferred from cell-based assays\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Established REC114 as a direct regulatory partner of TOPOVIBL within the SPO11/TOPOVIL catalytic complex, coupling the assembly platform to the nuclease.\",\n      \"evidence\": \"Structural domain mapping and TOPOVIBL point-mutant mice with genome-wide DSB mapping\",\n      \"pmids\": [\"36396648\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How TOPOVIBL binding triggers catalysis not resolved\", \"Temporal order relative to ANKRD31/IHO1 binding unclear\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Resolved the heterotrimer architecture and showed the REC114 PH-domain surface mediates mutually exclusive binding to IHO1, ANKRD31 and TOPOVIBL, recasting REC114 as a competitive regulatory hub.\",\n      \"evidence\": \"AlphaFold2 modeling validated by SEC, biochemistry and Co-IP\",\n      \"pmids\": [\"37431931\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"What controls partner exchange in vivo unknown\", \"Temporal regulation of competing interactions not defined\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined the structural basis of REC114-MEI4 DNA engagement, showing two oppositely oriented DNA-binding sites and preference for branched DNA that drive condensation.\",\n      \"evidence\": \"AlphaFold2, NMR, SAXS, mutagenesis and DNA-binding assays\",\n      \"pmids\": [\"37442581\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo relevance of branched-DNA preference not tested\", \"Link between condensation and Spo11 activation incomplete\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Demonstrated that a minimal REC114-MEI4 complex bridges and condenses DNA duplexes via long-range force, providing a biophysical mechanism for organizing DSB machinery.\",\n      \"evidence\": \"In vitro reconstitution with condensate and single-molecule force experiments\",\n      \"pmids\": [\"37442580\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological condensate composition not defined\", \"How bridging promotes Spo11 cleavage unresolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Showed genetically that the ANKRD31-REC114 interaction is required and dosage-sensitive for ANKRD31 scaffold function, validating the structural interface in vivo.\",\n      \"evidence\": \"Engineered Ankrd31 missense/truncation mice with DSB and crossover analysis\",\n      \"pmids\": [\"37976262\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometric basis of dosage sensitivity unresolved\", \"Whether REC114 limits or enables ANKRD31 at distinct loci unclear\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Confirmed in human males that a truncating REC114 variant disrupting MEI4 binding causes meiotic arrest and azoospermia, cementing the complex's role in human fertility.\",\n      \"evidence\": \"Co-IP, western blot, testicular histopathology and meiotic spreads from patients\",\n      \"pmids\": [\"38148155\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-family genetic evidence\", \"Quantitative effect on DSB numbers in patients not measured\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the competing PH-domain interactions, phosphorylation feedback, and condensate formation are temporally coordinated to license a precise number and distribution of DSBs remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No integrated in vivo timeline of partner exchange\", \"Mechanistic link between condensation and Spo11 catalysis missing\", \"Regulatory inputs governing DSB number not fully mapped\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [13, 14]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [8, 9, 12]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [11, 12]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005694\", \"supporting_discovery_ids\": [3, 4]},\n      {\"term_id\": \"GO:0000228\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1474165\", \"supporting_discovery_ids\": [8, 16]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 3]}\n    ],\n    \"complexes\": [\n      \"REC114-MEI4 heterotrimer\",\n      \"Mer2/IHO1-MEI4-REC114 pre-DSB complex\",\n      \"SPO11/TOPOVIL catalytic complex\"\n    ],\n    \"partners\": [\n      \"MEI4\",\n      \"IHO1\",\n      \"MER2\",\n      \"ANKRD31\",\n      \"TOPOVIBL\",\n      \"REC102\",\n      \"SPO11\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}