{"gene":"REC114","run_date":"2026-04-28T19:45:45","timeline":{"discoveries":[{"year":1993,"finding":"REC114 is a meiosis-specific gene required for meiotic recombination initiation, transcribed only during meiosis in an IME1-dependent manner, with no detectable function in mitosis.","method":"Genetic analysis, Northern blot hybridization, null mutant characterization","journal":"Current genetics","confidence":"Medium","confidence_rationale":"Tier 2 — genetic KO with defined meiotic phenotype, single lab","pmids":["8385581"],"is_preprint":false},{"year":1999,"finding":"High copy numbers of REC114 suppress meiotic DSB formation, indicating that overexpression of REC114 blocks double-strand break recombination intermediates in yeast.","method":"High-copy suppressor screen, genetic epistasis in dmc1 mutant background","journal":"Genes to cells","confidence":"Medium","confidence_rationale":"Tier 2 — genetic epistasis with defined DSB phenotype, 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 budding yeast, as shown by co-immunoprecipitation and co-localization on meiotic chromosome foci.","method":"Co-immunoprecipitation, immunofluorescence co-localization on meiotic chromosomes","journal":"Genetics","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP with co-localization, replicated across labs","pmids":["16783010"],"is_preprint":false},{"year":2007,"finding":"Rec114 localizes to meiotic chromosomes from leptonema through early pachynema, with localization independent of other DSB factors, and preferentially associates with chromatin loops rather than the meiotic cohesin Rec8.","method":"Immunofluorescence, chromosome spreads, genetic dependency analysis","journal":"Chromosoma","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization with functional genetic dependency analysis, single lab","pmids":["17558514"],"is_preprint":false},{"year":2007,"finding":"Mei4, Rec114, and Mer2 form a functional subgroup distinct from other DSB protein subgroups (Spo11-Ski8, Rec102-Rec104, and Mre11-Rad50-Xrs2); Rec102 and Rec104 connect Mei4 and Rec114 to Spo11.","method":"Two-hybrid interaction analysis, systematic deletion analysis, genetic epistasis","journal":"Chromosoma","confidence":"Medium","confidence_rationale":"Tier 2 — genetic epistasis and two-hybrid interaction mapping, single lab","pmids":["17558514"],"is_preprint":false},{"year":2007,"finding":"Spo11 self-interaction during meiosis is genetically regulated by Rec102, Rec104, and Rec114, as these proteins are required for Spo11 self-association at DSB sites.","method":"Co-immunoprecipitation of differentially tagged Spo11 proteins, chromatin analysis","journal":"Nucleic acids research","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP with genetic requirement established, single lab","pmids":["17264124"],"is_preprint":false},{"year":2013,"finding":"Tel1 (ATM) and Mec1 (ATR) phosphorylate Rec114 upon meiotic DSB formation, reducing Rec114 interaction with DSB hotspot DNA and down-regulating further DSB formation; non-phosphorylatable Rec114 causes genome-wide increase in DSBs.","method":"Phosphomimetic and non-phosphorylatable mutant alleles, chromatin immunoprecipitation, DSB quantification","journal":"PLoS genetics","confidence":"High","confidence_rationale":"Tier 1-2 — mutagenesis of phosphosites with multiple orthogonal readouts (ChIP, DSB levels), strong mechanistic evidence","pmids":["23825959"],"is_preprint":false},{"year":2013,"finding":"Ndt80, a meiosis-specific transcription factor, contributes to Rec114 degradation, providing a third independent mechanism for regulating Rec114 activity and DSB homeostasis.","method":"Genetic analysis in ndt80 mutant, protein level monitoring","journal":"PLoS genetics","confidence":"Medium","confidence_rationale":"Tier 2 — genetic KO 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 complex with REC114 and IHO1 in mouse spermatocytes; the REC114 C-terminal domain and MEI4 N-terminal domain form a stable complex in vitro; the REC114 N-terminal domain has structural similarity to Pleckstrin homology (PH) domains.","method":"Mouse KO, co-immunoprecipitation, in vitro reconstitution of REC114-MEI4 complex, crystal structure determination","journal":"Life science alliance","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution, structural analysis, and in vivo KO with defined phenotype in same study","pmids":["30569039"],"is_preprint":false},{"year":2019,"finding":"ANKRD31 directly interacts with REC114 via a pleckstrin homology (PH) domain in REC114, as defined by a crystal structure; ANKRD31 stabilizes REC114 association with the pseudoautosomal region and controls DSB number, timing, and location.","method":"Crystal structure of REC114 PH domain with ANKRD31, mouse KO, co-immunoprecipitation, cytological analysis","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1 — crystal structure with in vivo functional validation in mouse KO, multiple orthogonal methods","pmids":["31003867"],"is_preprint":false},{"year":2019,"finding":"A missense mutation (p.C133G) in REC114 reduces protein level and abolishes its function to protect MEI4 from degradation; a splice-site mutation affects alternative splicing of REC114; both cause multiple pronuclei formation and early embryonic arrest in human female infertility.","method":"Western blotting, co-immunoprecipitation, minigene splicing assay in HEK293T cells","journal":"Journal of medical genetics","confidence":"Medium","confidence_rationale":"Tier 2 — multiple biochemical methods in cell-based system, single lab","pmids":["31704776"],"is_preprint":false},{"year":2022,"finding":"REC114 is a direct binding partner of TOPOVIBL (the SPO11 partner in the TOPOVIL catalytic complex); their conserved interacting domains were identified by structural analysis; disruption of this interaction by point mutations in TOPOVIBL strongly reduces DSB activity genome-wide in oocytes and in sub-telomeric regions in spermatocytes.","method":"Structural analysis of REC114-TOPOVIBL interaction, point mutagenesis, meiotic DSB monitoring in mouse mutants","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1 — structural identification of interaction domain combined with in vivo mutagenesis and DSB quantification in mice","pmids":["36396648"],"is_preprint":false},{"year":2023,"finding":"Mouse REC114 forms homodimers and associates with MEI4 as a 2:1 heterotrimer that further dimerizes; IHO1 forms coiled-coil-based tetramers; IHO1 directly interacts with the PH domain of REC114 at the same surface recognized by TOPOVIBL and ANKRD31, suggesting REC114 acts as 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 — structural modeling validated by biochemical reconstitution with multiple orthogonal methods, single lab","pmids":["37431931"],"is_preprint":false},{"year":2023,"finding":"Rec114-Mei4 forms a 2:1 heterotrimeric complex where Rec114 C-terminus dimers cup an N-terminal Mei4 α-helix; this complex binds preferentially to branched DNA substrates via two DNA-binding sites that point in opposite directions and drives DNA condensation; the structure is conserved across eukaryotes.","method":"NMR spectroscopy, SAXS, mutagenesis, single-molecule experiments, AlphaFold2 modeling","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 1 — NMR, SAXS, and single-molecule experiments with mutagenesis in same study","pmids":["37442581","37442580"],"is_preprint":false},{"year":2023,"finding":"The minimal Rec114 C-terminus/Mei4 N-terminus heterotrimer is sufficient to bind DNA, bridge two or more DNA duplexes, and form large dynamic condensates; single-molecule experiments show the complex can condense DNA through long-range interactions.","method":"NMR, in vitro DNA binding assays, single-molecule experiments, condensate formation assays","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution with structural and single-molecule validation","pmids":["37442580"],"is_preprint":false},{"year":2023,"finding":"ANKRD31-REC114 interaction is essential for ANKRD31 function: complete loss of ANKRD31-REC114 interaction causes delayed DSB formation, defects in DSB repair, and failure to target DSBs to the pseudoautosomal regions; the severity of DSB defects correlates with the degree of disruption of this interaction.","method":"Mouse genetics with point mutations and truncations targeting ANKRD31-REC114 interface, DSB quantification, cytological analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 — multiple alleles with dosage effects and multiple phenotypic readouts, rigorous in vivo genetic dissection","pmids":["37976262"],"is_preprint":false},{"year":2023,"finding":"A truncating variant (p.Gln190*) in REC114 impairs interaction with MEI4, causing meiotic arrest and nonobstructive azoospermia in humans, confirming the REC114-MEI4 complex is essential for meiotic DSB homeostasis.","method":"Co-immunoprecipitation, Western blot in vitro, testicular histopathology, meiotic chromosome spread analysis","journal":"Clinical genetics","confidence":"Medium","confidence_rationale":"Tier 2 — biochemical validation of interaction disruption combined with histological meiotic arrest phenotype","pmids":["38148155"],"is_preprint":false},{"year":2025,"finding":"Mre11 condensates are recruited to meiotic DSB sites through interaction with Mer2 via a short α-helix in the Mre11 C-terminal IDR; Rec114-Mei4 and Mer2 (RMM) condensates organize the meiotic DSB machinery and are required for Mre11 recruitment.","method":"In vitro condensate assays, in vivo foci analysis, mutagenesis of Mre11-Mer2 interface, genetic complementation","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 — in vitro reconstitution with in vivo genetic validation, preprint not yet peer-reviewed","pmids":["bio_10.1101_2025.07.08.663703"],"is_preprint":true}],"current_model":"REC114 is an evolutionarily conserved meiotic factor that forms a 2:1 heterotrimer with MEI4 (with two REC114 subunits flanking one MEI4) and associates with IHO1/Mer2 to assemble a conserved DSB-promoting complex on meiotic chromosome axes; its N-terminal PH domain serves as a regulatory platform mediating mutually exclusive interactions with TOPOVIBL (the SPO11 catalytic partner), ANKRD31, and IHO1, thereby directly linking the axis-associated machinery to the DSB catalytic complex, while ATM/ATR-mediated phosphorylation of REC114 reduces its association with DSB hotspot chromatin to enforce DSB homeostasis."},"narrative":{"teleology":[{"year":1993,"claim":"Establishing that REC114 is a meiosis-specific gene required for recombination initiation answered whether REC114 functions exclusively in meiosis and placed it in the DSB-formation pathway.","evidence":"Null mutant characterization and Northern blot in budding yeast","pmids":["8385581"],"confidence":"Medium","gaps":["Molecular mechanism of how REC114 promotes DSBs unknown","Physical interaction partners not identified","No structural information available"]},{"year":2006,"claim":"Demonstrating that Rec114 forms a physical complex with Mer2 and Mei4 that co-localizes on meiotic chromosomes established a distinct protein subcomplex within the DSB machinery.","evidence":"Reciprocal co-immunoprecipitation and immunofluorescence co-localization in yeast","pmids":["16783010"],"confidence":"High","gaps":["Stoichiometry and architecture of the Rec114-Mei4-Mer2 complex unknown","How this complex connects to the Spo11 catalytic core not defined"]},{"year":2007,"claim":"Defining Rec114-Mei4-Mer2 as a functional subgroup separate from Spo11-Ski8 and identifying Rec102/Rec104 as connectors clarified the modular architecture of the DSB machinery and how Rec114 influences Spo11 self-association.","evidence":"Two-hybrid interaction mapping, genetic epistasis, and co-IP of differentially tagged Spo11 in yeast","pmids":["17558514","17264124"],"confidence":"Medium","gaps":["Direct structural basis of Rec114's contribution to Spo11 activity not resolved","Chromosome-loop versus axis association of Rec114 debated"]},{"year":2013,"claim":"Showing that ATM/ATR phosphorylate Rec114 to reduce its association with DSB hotspot DNA revealed a negative-feedback mechanism enforcing DSB homeostasis, answering how cells limit DSB numbers.","evidence":"Phosphomimetic/non-phosphorylatable mutants with ChIP and genome-wide DSB quantification in yeast","pmids":["23825959"],"confidence":"High","gaps":["Precise phosphosites mediating chromatin dissociation not fully mapped","Whether phosphorylation-dependent regulation is conserved in mammals untested at this stage","Ndt80-dependent degradation mechanism not biochemically defined"]},{"year":2018,"claim":"Demonstrating that mouse REC114 is essential for meiotic DSBs, forms a complex with MEI4 and IHO1, and possesses an N-terminal PH domain established evolutionary conservation and provided the first structural framework for REC114.","evidence":"Mouse knockout, in vitro reconstitution of REC114-MEI4 complex, crystal structure","pmids":["30569039"],"confidence":"High","gaps":["PH domain binding partners not yet identified","Stoichiometry of the REC114-MEI4 complex not resolved","How IHO1 connects the complex to the axis not structurally defined"]},{"year":2019,"claim":"Solving the crystal structure of the REC114 PH domain bound to ANKRD31 and showing ANKRD31 directs DSBs to the pseudoautosomal region via REC114 established the PH domain as a protein-interaction platform that controls DSB targeting.","evidence":"Crystal structure, mouse knockout of ANKRD31, co-IP, cytological analysis","pmids":["31003867"],"confidence":"High","gaps":["Whether additional partners engage the same PH domain surface unknown","Mechanism by which ANKRD31-REC114 interaction specifies PAR targeting not fully resolved"]},{"year":2022,"claim":"Identifying REC114 as a direct binding partner of TOPOVIBL and showing that disruption of this interaction impairs DSB activity answered how the axis-associated REC114 complex communicates with the SPO11 catalytic machinery.","evidence":"Structural analysis of REC114-TOPOVIBL domains, point mutagenesis with in vivo DSB monitoring in mouse mutants","pmids":["36396648"],"confidence":"High","gaps":["Full reconstitution of the complete SPO11-TOPOVIBL-REC114 assembly not achieved","Whether REC114-TOPOVIBL interaction is regulated by phosphorylation unknown"]},{"year":2023,"claim":"Resolving the 2:1 REC114-MEI4 heterotrimer architecture and demonstrating that IHO1, TOPOVIBL, and ANKRD31 compete for the same PH domain surface established REC114 as a regulatory switch coordinating axis association, DSB catalysis, and locus targeting through mutually exclusive interactions.","evidence":"AlphaFold2 modeling validated by SEC, co-IP, and biochemical reconstitution; NMR and SAXS of the heterotrimer; single-molecule DNA condensation assays; mouse point-mutant genetics for ANKRD31-REC114 interface","pmids":["37431931","37442581","37442580","37976262"],"confidence":"High","gaps":["How switching between PH-domain partners is temporally regulated in vivo is unknown","Whether DNA condensation by REC114-MEI4 is required for DSB formation in vivo untested","Full reconstitution of RMM condensates with SPO11 catalytic activity not achieved"]},{"year":2023,"claim":"Human genetic studies confirmed that loss-of-function REC114 mutations cause meiotic arrest, linking the REC114-MEI4 axis to nonobstructive azoospermia and female infertility with multiple pronuclei.","evidence":"Patient mutation analysis (p.Gln190*, p.C133G, splice-site variant), co-IP, histopathology","pmids":["38148155","31704776"],"confidence":"Medium","gaps":["Genotype-phenotype correlations across a larger patient cohort not established","Functional rescue experiments in human germline cells not performed"]},{"year":null,"claim":"Key open questions include how the temporal switching between mutually exclusive PH-domain partners is regulated in vivo, whether REC114-MEI4-driven DNA condensation is mechanistically required for DSB catalysis, and whether mammalian REC114 phosphorylation enforces DSB homeostasis as in yeast.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No reconstitution of SPO11-catalyzed DSB formation with purified REC114-MEI4-IHO1 complex","In vivo relevance of DNA condensation activity not tested","Mammalian phospho-regulation of REC114 not directly demonstrated"]}],"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":[9,11,12]}],"localization":[{"term_id":"GO:0005694","term_label":"chromosome","supporting_discovery_ids":[2,3,8]}],"pathway":[{"term_id":"R-HSA-1474165","term_label":"Reproduction","supporting_discovery_ids":[0,6,8,16]}],"complexes":["REC114-MEI4 heterotrimer","REC114-MEI4-IHO1/Mer2 (RMM) complex"],"partners":["MEI4","IHO1","ANKRD31","TOPOVIBL","MER2","SPO11"],"other_free_text":[]},"mechanistic_narrative":"REC114 is a meiosis-specific factor essential for the initiation of programmed DNA double-strand breaks (DSBs) that drive meiotic recombination. It forms a conserved 2:1 heterotrimer with MEI4, where two REC114 C-terminal domains cup an MEI4 N-terminal α-helix; this complex preferentially binds branched DNA and drives DNA condensation through multivalent bridging interactions [PMID:37442581, PMID:37442580, PMID:30569039]. The REC114 N-terminal PH domain serves as a regulatory hub mediating mutually exclusive interactions with IHO1/Mer2 (linking the complex to chromosome axes), ANKRD31 (directing DSBs to specific loci including pseudoautosomal regions), and TOPOVIBL (coupling the axis machinery to the SPO11 catalytic complex), while ATM/ATR-dependent phosphorylation of REC114 displaces it from hotspot chromatin to enforce DSB homeostasis [PMID:37431931, PMID:36396648, PMID:31003867, PMID:23825959]. Biallelic loss-of-function mutations in human REC114 cause meiotic arrest manifesting as nonobstructive azoospermia or female infertility with multiple pronuclei and early embryonic arrest [PMID:38148155, PMID:31704776]."},"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":110,"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":88,"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":79,"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":34,"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 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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":11177,"output_tokens":3703,"usd":0.044538},"stage2":{"model":"claude-opus-4-6","input_tokens":7163,"output_tokens":2587,"usd":0.150735},"total_usd":0.195273,"stage1_batch_id":"msgbatch_01SUR8NDzVrNmyszWyJeCct7","stage2_batch_id":"msgbatch_01G7Yfutko3cDWzejrwH5AMx","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1993,\n      \"finding\": \"REC114 is a meiosis-specific gene required for meiotic recombination initiation, transcribed only during meiosis in an IME1-dependent manner, with no detectable function in mitosis.\",\n      \"method\": \"Genetic analysis, Northern blot hybridization, null mutant characterization\",\n      \"journal\": \"Current genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO with defined meiotic phenotype, single lab\",\n      \"pmids\": [\"8385581\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"High copy numbers of REC114 suppress meiotic DSB formation, indicating that overexpression of REC114 blocks double-strand break recombination intermediates in yeast.\",\n      \"method\": \"High-copy suppressor screen, genetic epistasis in dmc1 mutant background\",\n      \"journal\": \"Genes to cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis with defined DSB phenotype, 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 budding yeast, as shown by co-immunoprecipitation and co-localization on meiotic chromosome foci.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence co-localization on meiotic chromosomes\",\n      \"journal\": \"Genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP with co-localization, replicated across labs\",\n      \"pmids\": [\"16783010\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Rec114 localizes to meiotic chromosomes from leptonema through early pachynema, with localization independent of other DSB factors, and preferentially associates with chromatin loops rather than the meiotic cohesin Rec8.\",\n      \"method\": \"Immunofluorescence, chromosome spreads, genetic dependency analysis\",\n      \"journal\": \"Chromosoma\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization with functional genetic dependency analysis, single lab\",\n      \"pmids\": [\"17558514\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Mei4, Rec114, and Mer2 form a functional subgroup distinct from other DSB protein subgroups (Spo11-Ski8, Rec102-Rec104, and Mre11-Rad50-Xrs2); Rec102 and Rec104 connect Mei4 and Rec114 to Spo11.\",\n      \"method\": \"Two-hybrid interaction analysis, systematic deletion analysis, genetic epistasis\",\n      \"journal\": \"Chromosoma\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis and two-hybrid interaction mapping, single lab\",\n      \"pmids\": [\"17558514\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Spo11 self-interaction during meiosis is genetically regulated by Rec102, Rec104, and Rec114, as these proteins are required for Spo11 self-association at DSB sites.\",\n      \"method\": \"Co-immunoprecipitation of differentially tagged Spo11 proteins, chromatin analysis\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP with genetic requirement established, single lab\",\n      \"pmids\": [\"17264124\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Tel1 (ATM) and Mec1 (ATR) phosphorylate Rec114 upon meiotic DSB formation, reducing Rec114 interaction with DSB hotspot DNA and down-regulating further DSB formation; non-phosphorylatable Rec114 causes genome-wide increase in DSBs.\",\n      \"method\": \"Phosphomimetic and non-phosphorylatable mutant alleles, chromatin immunoprecipitation, DSB quantification\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — mutagenesis of phosphosites with multiple orthogonal readouts (ChIP, DSB levels), strong mechanistic evidence\",\n      \"pmids\": [\"23825959\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Ndt80, a meiosis-specific transcription factor, contributes to Rec114 degradation, providing a third independent mechanism for regulating Rec114 activity and DSB homeostasis.\",\n      \"method\": \"Genetic analysis in ndt80 mutant, protein level monitoring\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO 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 complex with REC114 and IHO1 in mouse spermatocytes; the REC114 C-terminal domain and MEI4 N-terminal domain form a stable complex in vitro; the REC114 N-terminal domain has structural similarity to Pleckstrin homology (PH) domains.\",\n      \"method\": \"Mouse KO, co-immunoprecipitation, in vitro reconstitution of REC114-MEI4 complex, crystal structure determination\",\n      \"journal\": \"Life science alliance\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution, structural analysis, and in vivo KO with defined phenotype in same study\",\n      \"pmids\": [\"30569039\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"ANKRD31 directly interacts with REC114 via a pleckstrin homology (PH) domain in REC114, as defined by a crystal structure; ANKRD31 stabilizes REC114 association with the pseudoautosomal region and controls DSB number, timing, and location.\",\n      \"method\": \"Crystal structure of REC114 PH domain with ANKRD31, mouse KO, co-immunoprecipitation, cytological analysis\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — crystal structure with in vivo functional validation in mouse KO, multiple orthogonal methods\",\n      \"pmids\": [\"31003867\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"A missense mutation (p.C133G) in REC114 reduces protein level and abolishes its function to protect MEI4 from degradation; a splice-site mutation affects alternative splicing of REC114; both cause multiple pronuclei formation and early embryonic arrest in human female infertility.\",\n      \"method\": \"Western blotting, co-immunoprecipitation, minigene splicing assay in HEK293T cells\",\n      \"journal\": \"Journal of medical genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple biochemical methods in cell-based system, single lab\",\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 TOPOVIL catalytic complex); their conserved interacting domains were identified by structural analysis; disruption of this interaction by point mutations in TOPOVIBL strongly reduces DSB activity genome-wide in oocytes and in sub-telomeric regions in spermatocytes.\",\n      \"method\": \"Structural analysis of REC114-TOPOVIBL interaction, point mutagenesis, meiotic DSB monitoring in mouse mutants\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — structural identification of interaction domain combined with in vivo mutagenesis and DSB quantification in mice\",\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 heterotrimer that further dimerizes; IHO1 forms coiled-coil-based tetramers; IHO1 directly interacts with the PH domain of REC114 at the same surface recognized by TOPOVIBL and ANKRD31, suggesting REC114 acts as 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 — structural modeling validated by biochemical reconstitution with multiple orthogonal methods, single lab\",\n      \"pmids\": [\"37431931\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Rec114-Mei4 forms a 2:1 heterotrimeric complex where Rec114 C-terminus dimers cup an N-terminal Mei4 α-helix; this complex binds preferentially to branched DNA substrates via two DNA-binding sites that point in opposite directions and drives DNA condensation; the structure is conserved across eukaryotes.\",\n      \"method\": \"NMR spectroscopy, SAXS, mutagenesis, single-molecule experiments, AlphaFold2 modeling\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — NMR, SAXS, and single-molecule experiments with mutagenesis in same study\",\n      \"pmids\": [\"37442581\", \"37442580\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The minimal Rec114 C-terminus/Mei4 N-terminus heterotrimer is sufficient to bind DNA, bridge two or more DNA duplexes, and form large dynamic condensates; single-molecule experiments show the complex can condense DNA through long-range interactions.\",\n      \"method\": \"NMR, in vitro DNA binding assays, single-molecule experiments, condensate formation assays\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution with structural and single-molecule validation\",\n      \"pmids\": [\"37442580\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ANKRD31-REC114 interaction is essential for ANKRD31 function: complete loss of ANKRD31-REC114 interaction causes delayed DSB formation, defects in DSB repair, and failure to target DSBs to the pseudoautosomal regions; the severity of DSB defects correlates with the degree of disruption of this interaction.\",\n      \"method\": \"Mouse genetics with point mutations and truncations targeting ANKRD31-REC114 interface, DSB quantification, cytological analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple alleles with dosage effects and multiple phenotypic readouts, rigorous in vivo genetic dissection\",\n      \"pmids\": [\"37976262\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"A truncating variant (p.Gln190*) in REC114 impairs interaction with MEI4, causing meiotic arrest and nonobstructive azoospermia in humans, confirming the REC114-MEI4 complex is essential for meiotic DSB homeostasis.\",\n      \"method\": \"Co-immunoprecipitation, Western blot in vitro, testicular histopathology, meiotic chromosome spread analysis\",\n      \"journal\": \"Clinical genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — biochemical validation of interaction disruption combined with histological meiotic arrest phenotype\",\n      \"pmids\": [\"38148155\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Mre11 condensates are recruited to meiotic DSB sites through interaction with Mer2 via a short α-helix in the Mre11 C-terminal IDR; Rec114-Mei4 and Mer2 (RMM) condensates organize the meiotic DSB machinery and are required for Mre11 recruitment.\",\n      \"method\": \"In vitro condensate assays, in vivo foci analysis, mutagenesis of Mre11-Mer2 interface, genetic complementation\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vitro reconstitution with in vivo genetic validation, preprint not yet peer-reviewed\",\n      \"pmids\": [\"bio_10.1101_2025.07.08.663703\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"REC114 is an evolutionarily conserved meiotic factor that forms a 2:1 heterotrimer with MEI4 (with two REC114 subunits flanking one MEI4) and associates with IHO1/Mer2 to assemble a conserved DSB-promoting complex on meiotic chromosome axes; its N-terminal PH domain serves as a regulatory platform mediating mutually exclusive interactions with TOPOVIBL (the SPO11 catalytic partner), ANKRD31, and IHO1, thereby directly linking the axis-associated machinery to the DSB catalytic complex, while ATM/ATR-mediated phosphorylation of REC114 reduces its association with DSB hotspot chromatin to enforce DSB homeostasis.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"REC114 is a meiosis-specific factor essential for the initiation of programmed DNA double-strand breaks (DSBs) that drive meiotic recombination. It forms a conserved 2:1 heterotrimer with MEI4, where two REC114 C-terminal domains cup an MEI4 N-terminal α-helix; this complex preferentially binds branched DNA and drives DNA condensation through multivalent bridging interactions [PMID:37442581, PMID:37442580, PMID:30569039]. The REC114 N-terminal PH domain serves as a regulatory hub mediating mutually exclusive interactions with IHO1/Mer2 (linking the complex to chromosome axes), ANKRD31 (directing DSBs to specific loci including pseudoautosomal regions), and TOPOVIBL (coupling the axis machinery to the SPO11 catalytic complex), while ATM/ATR-dependent phosphorylation of REC114 displaces it from hotspot chromatin to enforce DSB homeostasis [PMID:37431931, PMID:36396648, PMID:31003867, PMID:23825959]. Biallelic loss-of-function mutations in human REC114 cause meiotic arrest manifesting as nonobstructive azoospermia or female infertility with multiple pronuclei and early embryonic arrest [PMID:38148155, PMID:31704776].\",\n  \"teleology\": [\n    {\n      \"year\": 1993,\n      \"claim\": \"Establishing that REC114 is a meiosis-specific gene required for recombination initiation answered whether REC114 functions exclusively in meiosis and placed it in the DSB-formation pathway.\",\n      \"evidence\": \"Null mutant characterization and Northern blot in budding yeast\",\n      \"pmids\": [\"8385581\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Molecular mechanism of how REC114 promotes DSBs unknown\",\n        \"Physical interaction partners not identified\",\n        \"No structural information available\"\n      ]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Demonstrating that Rec114 forms a physical complex with Mer2 and Mei4 that co-localizes on meiotic chromosomes established a distinct protein subcomplex within the DSB machinery.\",\n      \"evidence\": \"Reciprocal co-immunoprecipitation and immunofluorescence co-localization in yeast\",\n      \"pmids\": [\"16783010\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Stoichiometry and architecture of the Rec114-Mei4-Mer2 complex unknown\",\n        \"How this complex connects to the Spo11 catalytic core not defined\"\n      ]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Defining Rec114-Mei4-Mer2 as a functional subgroup separate from Spo11-Ski8 and identifying Rec102/Rec104 as connectors clarified the modular architecture of the DSB machinery and how Rec114 influences Spo11 self-association.\",\n      \"evidence\": \"Two-hybrid interaction mapping, genetic epistasis, and co-IP of differentially tagged Spo11 in yeast\",\n      \"pmids\": [\"17558514\", \"17264124\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct structural basis of Rec114's contribution to Spo11 activity not resolved\",\n        \"Chromosome-loop versus axis association of Rec114 debated\"\n      ]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Showing that ATM/ATR phosphorylate Rec114 to reduce its association with DSB hotspot DNA revealed a negative-feedback mechanism enforcing DSB homeostasis, answering how cells limit DSB numbers.\",\n      \"evidence\": \"Phosphomimetic/non-phosphorylatable mutants with ChIP and genome-wide DSB quantification in yeast\",\n      \"pmids\": [\"23825959\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Precise phosphosites mediating chromatin dissociation not fully mapped\",\n        \"Whether phosphorylation-dependent regulation is conserved in mammals untested at this stage\",\n        \"Ndt80-dependent degradation mechanism not biochemically defined\"\n      ]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Demonstrating that mouse REC114 is essential for meiotic DSBs, forms a complex with MEI4 and IHO1, and possesses an N-terminal PH domain established evolutionary conservation and provided the first structural framework for REC114.\",\n      \"evidence\": \"Mouse knockout, in vitro reconstitution of REC114-MEI4 complex, crystal structure\",\n      \"pmids\": [\"30569039\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"PH domain binding partners not yet identified\",\n        \"Stoichiometry of the REC114-MEI4 complex not resolved\",\n        \"How IHO1 connects the complex to the axis not structurally defined\"\n      ]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Solving the crystal structure of the REC114 PH domain bound to ANKRD31 and showing ANKRD31 directs DSBs to the pseudoautosomal region via REC114 established the PH domain as a protein-interaction platform that controls DSB targeting.\",\n      \"evidence\": \"Crystal structure, mouse knockout of ANKRD31, co-IP, cytological analysis\",\n      \"pmids\": [\"31003867\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether additional partners engage the same PH domain surface unknown\",\n        \"Mechanism by which ANKRD31-REC114 interaction specifies PAR targeting not fully resolved\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identifying REC114 as a direct binding partner of TOPOVIBL and showing that disruption of this interaction impairs DSB activity answered how the axis-associated REC114 complex communicates with the SPO11 catalytic machinery.\",\n      \"evidence\": \"Structural analysis of REC114-TOPOVIBL domains, point mutagenesis with in vivo DSB monitoring in mouse mutants\",\n      \"pmids\": [\"36396648\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Full reconstitution of the complete SPO11-TOPOVIBL-REC114 assembly not achieved\",\n        \"Whether REC114-TOPOVIBL interaction is regulated by phosphorylation unknown\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Resolving the 2:1 REC114-MEI4 heterotrimer architecture and demonstrating that IHO1, TOPOVIBL, and ANKRD31 compete for the same PH domain surface established REC114 as a regulatory switch coordinating axis association, DSB catalysis, and locus targeting through mutually exclusive interactions.\",\n      \"evidence\": \"AlphaFold2 modeling validated by SEC, co-IP, and biochemical reconstitution; NMR and SAXS of the heterotrimer; single-molecule DNA condensation assays; mouse point-mutant genetics for ANKRD31-REC114 interface\",\n      \"pmids\": [\"37431931\", \"37442581\", \"37442580\", \"37976262\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"How switching between PH-domain partners is temporally regulated in vivo is unknown\",\n        \"Whether DNA condensation by REC114-MEI4 is required for DSB formation in vivo untested\",\n        \"Full reconstitution of RMM condensates with SPO11 catalytic activity not achieved\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Human genetic studies confirmed that loss-of-function REC114 mutations cause meiotic arrest, linking the REC114-MEI4 axis to nonobstructive azoospermia and female infertility with multiple pronuclei.\",\n      \"evidence\": \"Patient mutation analysis (p.Gln190*, p.C133G, splice-site variant), co-IP, histopathology\",\n      \"pmids\": [\"38148155\", \"31704776\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Genotype-phenotype correlations across a larger patient cohort not established\",\n        \"Functional rescue experiments in human germline cells not performed\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key open questions include how the temporal switching between mutually exclusive PH-domain partners is regulated in vivo, whether REC114-MEI4-driven DNA condensation is mechanistically required for DSB catalysis, and whether mammalian REC114 phosphorylation enforces DSB homeostasis as in yeast.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No reconstitution of SPO11-catalyzed DSB formation with purified REC114-MEI4-IHO1 complex\",\n        \"In vivo relevance of DNA condensation activity not tested\",\n        \"Mammalian phospho-regulation of REC114 not directly demonstrated\"\n      ]\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\": [9, 11, 12]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005694\", \"supporting_discovery_ids\": [2, 3, 8]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1474165\", \"supporting_discovery_ids\": [0, 6, 8, 16]}\n    ],\n    \"complexes\": [\n      \"REC114-MEI4 heterotrimer\",\n      \"REC114-MEI4-IHO1/Mer2 (RMM) complex\"\n    ],\n    \"partners\": [\n      \"MEI4\",\n      \"IHO1\",\n      \"ANKRD31\",\n      \"TOPOVIBL\",\n      \"MER2\",\n      \"SPO11\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}