{"gene":"ACTL7A","run_date":"2026-06-09T22:02:40","timeline":{"discoveries":[{"year":1999,"finding":"ACTL7A encodes a 435-amino-acid actin-like protein (predicted MW 48.6 kDa) expressed in multiple adult tissues, with the gene located intronless on chromosome 9q31 in a head-to-head orientation with ACTL7B on a common 8-kb HindIII fragment.","method":"cDNA selection, direct genomic sequencing, linkage analysis","journal":"Genomics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct cloning and genomic characterization with expression analysis across tissues, single lab, multiple methods","pmids":["10373328"],"is_preprint":false},{"year":2012,"finding":"ACTL7A expression is upregulated via the PKA pathway and undergoes relocalization (remodeling) during the early period of capacitation in mouse spermatozoa, indicating it is an essential component of sperm capacitation.","method":"Western blot, indirect immunostaining during capacitation induction","journal":"Fertility and sterility","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — two methods (western blot + immunostaining), single lab, no genetic rescue or in vitro reconstitution","pmids":["23211711"],"is_preprint":false},{"year":2012,"finding":"Anti-ACTL7A antibodies cause sperm agglutination and reduce fertilizing capacity of mouse spermatozoa in vitro; active immunization of mice with ACTL7A protein significantly reduces fertility, establishing ACTL7A as a target antigen in immunologic infertility.","method":"In vitro sperm treatment with antibody-containing serum, active immunization fertility assay, sperm agglutination test, mass spectrometry identification","journal":"Fertility and sterility","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional fertility assay in vivo and in vitro, multiple orthogonal methods, single lab","pmids":["22386842"],"is_preprint":false},{"year":2020,"finding":"A homozygous missense mutation in ACTL7A causes acrosomal ultrastructural defects in sperm and leads to reduced expression and abnormal localization of PLCζ in sperm, resulting in failure of oocyte activation and early embryonic arrest. Artificial oocyte activation rescues the fertilization defect.","method":"Whole-exome sequencing, knock-in mouse model, transmission electron microscopy, immunofluorescence, western blot","journal":"Science advances","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic patient data corroborated by knock-in mouse model with consistent phenotype, multiple orthogonal methods (TEM, IF, WB), functional rescue by AOA","pmids":["32923619"],"is_preprint":false},{"year":2021,"finding":"Compound heterozygous loss-of-function variants in ACTL7A cause ultrastructural defects in the acrosome and perinuclear theca, and significantly reduce expression of both ACTL7A protein and PLCζ in sperm, leading to oocyte activation deficiency and total fertilization failure.","method":"Whole-exome sequencing, Sanger sequencing, transmission electron microscopy, immunofluorescence, western blot","journal":"Human reproduction","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods, single family/lab, replicates mechanism from prior study","pmids":["34727571"],"is_preprint":false},{"year":2022,"finding":"In Actl7a knockout mice, sperm show malformed acrosomes, altered localization of zona pellucida binding protein ZPBP, and reduced calcium oscillations in oocytes due to abnormal localization and expression of PLCZ1; ACTL7A and ZPBP co-immunoprecipitate, forming a complex potentially involved in acrosomal formation.","method":"Knockout mouse model, immunofluorescence, Co-immunoprecipitation, western blot, IVF/ICSI","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO mouse with defined phenotype plus Co-IP for complex formation, single lab","pmids":["35921706"],"is_preprint":false},{"year":2022,"finding":"A pathogenic variant in ACTL7A (p.Gly402Ser) causes mutant ACTL7A to fail to attach to the acroplaxome and be discharged via cytoplasmic droplets, resulting in absence of ACTL7A from epididymal sperm, acrosome detachment from the nuclear membrane (bubble-shaped acrosomes), and PLCζ co-discharge leading to total fertilization failure. Immunoprecipitation-mass spectrometry identified interacting proteins involved in acrosome assembly and actin filament organization.","method":"Knock-in mouse model (equivalent variant), transmission electron microscopy, immunofluorescence, immunoprecipitation/LC-MS, western blot","journal":"Molecular human reproduction","confidence":"High","confidence_rationale":"Tier 2 / Strong — knock-in mouse model replicating patient phenotype, multiple orthogonal methods including IP-MS for interactome, mechanistic explanation of protein mislocalization and discharge","pmids":["35863052"],"is_preprint":false},{"year":2022,"finding":"A homozygous missense mutation p.D75A in ACTL7A causes protein degradation in sperm, irregular perinuclear theca and acrosomal ultrastructural defects, and abnormal localization and reduced expression of PLCZ1; 3D structural modeling shows loss of a hydrogen bond with Ser170 and transformation of an α-helix to random coil.","method":"Whole-exome sequencing, 3D structural modeling, immunofluorescence, transmission electron microscopy, western blot","journal":"Molecular genetics and genomics","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — multiple clinical and experimental methods, structural modeling is in silico only, single lab","pmids":["36574082"],"is_preprint":false},{"year":2023,"finding":"ACTL7A is dynamically localized within the nucleus and subacrosomal space of developing spermatids, and later associates with postacrosomal regions. Actl7a knockout mice show complete loss of subacrosomal filamentous actin (F-actin) structures, abnormal acrosomal granule migration, and peeling acrosomes during spermatid elongation, establishing ACTL7A as required for subacrosomal F-actin formation and acrosomal anchoring via the acroplaxome.","method":"Actl7a knockout mouse model, immunofluorescence (dynamic localization), phalloidin staining for F-actin, transmission electron microscopy","journal":"Molecular human reproduction","confidence":"High","confidence_rationale":"Tier 2 / Strong — KO mouse model with multiple orthogonal methods, direct demonstration of F-actin loss as mechanistic link, single lab with rigorous controls","pmids":["36734600"],"is_preprint":false},{"year":2023,"finding":"Loss of ACTL7A disrupts the acrosome-acroplaxome-manchette complex, causing small head sperm. Proteomic analysis of Actl7a-KO testes reveals enrichment of differentially expressed proteins in the PI3K/AKT/mTOR pathway; autophagy inhibition via PI3K/AKT/mTOR activation leads to PDLIM1 accumulation, impairing manchette development and sperm head shaping.","method":"Actl7a knockout mouse model, immunofluorescence, transmission electron microscopy, tandem mass tag quantitative proteomics, western blot","journal":"Reproductive biology and endocrinology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO mouse with proteomics and multiple validation methods, PI3K/AKT/mTOR/autophagy pathway placement, single lab","pmids":["37667331"],"is_preprint":false},{"year":2023,"finding":"ACTL7A variants affecting the actin domain cause absent ACTL7A protein in spermatozoa and near-absent PLCζ1, along with attenuated and unevenly distributed acrosomal PNA signals, indicating acrosome dysfunction and oocyte activation failure.","method":"Whole-exome sequencing, immunofluorescence, western blot, bioinformatic structural prediction","journal":"Andrology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — multiple patient cases with consistent findings, immunofluorescence and WB, no KO model, single lab","pmids":["37991128"],"is_preprint":false},{"year":2024,"finding":"In the absence of ACTL7A (or ACTL7B), intranuclear localization of HDAC1 and HDAC3 is lost in spermatids, implicating ACTL7A in nuclear HDAC association and epigenetic regulation during spermiogenesis. In silico modeling predicts ACTL7A can bind to HSA domains of INO80 and SWI/SNF nucleosome remodeler family members in a manner analogous to nuclear actin and ACTL6A, suggesting ARP subunit swapping in chromatin regulatory complexes.","method":"Actl7a and Actl7b knockout mouse models, immunofluorescence for HDAC1/HDAC3 localization, transcriptomic analysis, in silico structural modeling","journal":"bioRxiv","confidence":"Low","confidence_rationale":"Tier 3 / Weak — preprint, HDAC mislocalization shown by IF in KO mice (moderate evidence) but nucleosome remodeler interactions are in silico predictions only, not biochemically validated","pmids":["38464253"],"is_preprint":true},{"year":2025,"finding":"FNDC8 interacts with ACTL7A (and CCIN) in the perinuclear theca during spermiogenesis; depletion of FNDC8 destabilizes ACTL7A protein levels, establishing ACTL7A as part of a protein complex within the perinuclear theca that maintains structural integrity for sperm head morphogenesis.","method":"Co-immunoprecipitation, Fndc8 knockout mouse model, immunofluorescence, western blot","journal":"Zoological research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP demonstrating interaction, KO mouse showing destabilization of ACTL7A, single lab","pmids":["41169243"],"is_preprint":false},{"year":2025,"finding":"ACTL7A co-immunoprecipitates with the perinuclear theca proteins ACTRT1, ACTRT2, and ARPM1, and with the sperm surface protein ZPBP, placing ACTL7A within a cytoskeletal network in the perinuclear theca that connects the acrosome and nucleus.","method":"Co-immunoprecipitation in Arpm1-deficient mouse model context","journal":"bioRxiv","confidence":"Low","confidence_rationale":"Tier 3 / Weak — preprint, single Co-IP experiment, ACTL7A is not the primary subject of the study","pmids":["bio_10.1101_2025.03.27.645694"],"is_preprint":true}],"current_model":"ACTL7A is a testis-enriched actin-like protein that localizes dynamically to the nucleus, subacrosomal space, and postacrosomal regions of developing spermatids, where it is required for subacrosomal F-actin formation, acroplaxome-mediated acrosomal attachment to the nucleus, and sperm head shaping; loss-of-function mutations cause acrosome detachment, perinuclear theca defects, and mislocalization/loss of PLCζ (PLCZ1), the sperm-borne oocyte activation factor, resulting in oocyte activation failure, total fertilization failure, and male infertility that can be rescued by artificial oocyte activation."},"narrative":{"mechanistic_narrative":"ACTL7A is a testis-enriched actin-like protein essential for acrosome biogenesis and sperm head shaping during spermiogenesis [PMID:36734600]. It localizes dynamically to the nucleus and subacrosomal space of developing spermatids and later to postacrosomal regions, where it drives formation of subacrosomal filamentous actin and anchors the acrosome to the nucleus via the acroplaxome; its loss abolishes subacrosomal F-actin, causes abnormal acrosomal granule migration, and produces peeling, detached acrosomes [PMID:36734600]. ACTL7A operates within a perinuclear theca cytoskeletal network, co-immunoprecipitating with the zona-pellucida-binding protein ZPBP and being stabilized by FNDC8, whose depletion destabilizes ACTL7A and disrupts head morphogenesis [PMID:35921706, PMID:41169243]. Disruption of the acrosome-acroplaxome-manchette complex upon ACTL7A loss yields small-headed sperm, accompanied by dysregulation of the PI3K/AKT/mTOR/autophagy axis and PDLIM1 accumulation that impairs manchette development [PMID:37667331]. A central downstream consequence of ACTL7A dysfunction is mislocalization, reduced expression, or co-discharge of the sperm-borne oocyte activation factor PLCζ (PLCZ1), which abolishes oocyte calcium oscillations and causes total fertilization failure [PMID:32923619, PMID:35863052]. Loss-of-function and missense mutations in ACTL7A cause male infertility through acrosomal and perinuclear theca ultrastructural defects and oocyte activation deficiency, a phenotype rescuable by artificial oocyte activation [PMID:32923619, PMID:34727571].","teleology":[{"year":1999,"claim":"Established ACTL7A's molecular identity by cloning it as an intronless actin-like gene paired head-to-head with ACTL7B, defining the basic gene and protein before any functional role was known.","evidence":"cDNA selection and direct genomic sequencing with multi-tissue expression analysis","pmids":["10373328"],"confidence":"Medium","gaps":["No protein function or subcellular role defined","Testis-specific role not yet identified"]},{"year":2012,"claim":"First linked ACTL7A to sperm physiology by showing PKA-driven upregulation and relocalization during capacitation, and demonstrating that antibody targeting impairs fertilization, framing it as a functionally important and immunologically accessible sperm antigen.","evidence":"Western blot and immunostaining during capacitation; in vitro sperm antibody treatment and active immunization fertility assays in mouse","pmids":["23211711","22386842"],"confidence":"Medium","gaps":["Molecular mechanism of relocalization unknown","No genetic loss-of-function evidence","Direct binding partners not identified"]},{"year":2020,"claim":"Connected ACTL7A mutation to human infertility by showing a missense variant causes acrosomal defects and PLCζ mislocalization that block oocyte activation, with rescue by artificial oocyte activation establishing the therapeutic and mechanistic link.","evidence":"Whole-exome sequencing, knock-in mouse, TEM, immunofluorescence, western blot, AOA rescue","pmids":["32923619"],"confidence":"High","gaps":["How ACTL7A controls PLCζ localization not resolved","Structural basis of the missense effect not defined"]},{"year":2021,"claim":"Confirmed and broadened the disease link by showing biallelic loss-of-function variants reduce ACTL7A and PLCζ and produce perinuclear theca defects, reinforcing oocyte activation deficiency as the core pathology.","evidence":"Whole-exome and Sanger sequencing, TEM, immunofluorescence, western blot in patient sperm","pmids":["34727571"],"confidence":"Medium","gaps":["No mouse model in this study","Mechanistic coupling of PT integrity to PLCζ retention unclear"]},{"year":2022,"claim":"Defined ACTL7A's physical partnerships and the consequence of mislocalization, showing it complexes with ZPBP and that variant protein fails to attach to the acroplaxome and is discharged with PLCζ, explaining the loss of both proteins from sperm.","evidence":"Knockout and knock-in mouse models, Co-IP, IP-LC/MS interactome, TEM, immunofluorescence, IVF/ICSI","pmids":["35921706","35863052"],"confidence":"High","gaps":["Stoichiometry and direct vs indirect nature of interactions not resolved","How acroplaxome attachment is molecularly mediated unknown"]},{"year":2022,"claim":"Provided structural rationale for a pathogenic missense allele, showing p.D75A destabilizes ACTL7A via loss of a hydrogen bond and helix-to-coil transition, linking protein folding to PT/acrosome defects and PLCZ1 dysregulation.","evidence":"Whole-exome sequencing, in silico 3D structural modeling, immunofluorescence, TEM, western blot","pmids":["36574082"],"confidence":"Medium","gaps":["Structural prediction not experimentally validated","Degradation pathway not identified"]},{"year":2023,"claim":"Identified the core cell-biological function as subacrosomal F-actin formation and acroplaxome-mediated acrosomal anchoring, the mechanistic basis for acrosome attachment that earlier mutation studies implicated indirectly.","evidence":"Actl7a knockout mouse, phalloidin F-actin staining, dynamic-localization immunofluorescence, TEM","pmids":["36734600"],"confidence":"High","gaps":["Biochemical mechanism of F-actin nucleation/organization by ACTL7A unknown","Whether ACTL7A polymerizes or scaffolds conventional actin unresolved"]},{"year":2023,"claim":"Placed ACTL7A within a broader head-shaping program, linking its loss to acrosome-acroplaxome-manchette disruption and to PI3K/AKT/mTOR-mediated autophagy inhibition and PDLIM1 accumulation that impairs the manchette.","evidence":"Actl7a knockout mouse, TMT quantitative proteomics, immunofluorescence, TEM, western blot","pmids":["37667331"],"confidence":"Medium","gaps":["Causality between ACTL7A loss and pathway activation not directly tested","Direct vs secondary effect on manchette unclear"]},{"year":2023,"claim":"Extended the human genetic spectrum to actin-domain variants, confirming absent sperm ACTL7A, near-absent PLCζ, and impaired acrosomal PNA staining as a reproducible patient phenotype.","evidence":"Whole-exome sequencing, immunofluorescence, western blot, bioinformatic structural prediction in patients","pmids":["37991128"],"confidence":"Medium","gaps":["No functional model in this cohort","Genotype-phenotype correlation across variants incomplete"]},{"year":2025,"claim":"Defined a perinuclear theca complex by showing FNDC8 interacts with and stabilizes ACTL7A, providing a mechanism for maintaining ACTL7A protein levels needed for head morphogenesis.","evidence":"Co-immunoprecipitation and Fndc8 knockout mouse, immunofluorescence, western blot","pmids":["41169243"],"confidence":"Medium","gaps":["Whether stabilization is direct or via complex assembly unknown","Other PT partners maintaining ACTL7A not fully mapped"]},{"year":2025,"claim":"Began mapping a wider PT cytoskeletal network by Co-IP of ACTL7A with ACTRT1/ACTRT2/ARPM1 and ZPBP, and proposed a nuclear chromatin-regulatory role via HDAC1/HDAC3 association and putative ARP subunit swapping in remodeler complexes.","evidence":"Co-IP in Arpm1-deficient context (preprint); Actl7a/Actl7b knockout mice with HDAC1/HDAC3 immunofluorescence, transcriptomics, in silico modeling (preprint)","pmids":["bio_10.1101_2025.03.27.645694","38464253"],"confidence":"Low","gaps":["Single Co-IP without reciprocal validation; ACTL7A not the primary subject","Nucleosome-remodeler interactions are in silico predictions only, not biochemically validated","Nuclear/chromatin function not connected to the cytoplasmic acrosomal role"]},{"year":null,"claim":"The biochemical mechanism by which ACTL7A nucleates or organizes subacrosomal F-actin, and how this physically retains PLCζ in the perinuclear theca, remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No in vitro reconstitution of ACTL7A actin activity","Direct ACTL7A-PLCζ interaction not demonstrated","Structural model of ACTL7A in the acroplaxome lacking"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[8]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[8,13]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[8,11]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[8,13]}],"pathway":[{"term_id":"R-HSA-1474165","term_label":"Reproduction","supporting_discovery_ids":[3,8]}],"complexes":["acroplaxome","perinuclear theca cytoskeletal network","acrosome-acroplaxome-manchette complex"],"partners":["ZPBP","FNDC8","CCIN","ACTRT1","ACTRT2","ARPM1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9Y615","full_name":"Actin-like protein 7A","aliases":["Actin-like-7-alpha"],"length_aa":435,"mass_kda":48.6,"function":"Essential for normal spermatogenesis and male fertility. Required for normal sperm head morphology, acroplaxome formation, acrosome attachment, and acrosome granule stability. May anchor and stabilize acrosomal adherence to the acroplaxome at least in part by facilitating the presence of F-actin in the subacrosomal space (By similarity). May play an important role in formation and fusion of Golgi-derived vesicles during acrosome biogenesis (PubMed:32923619)","subcellular_location":"Cytoplasm, cytoskeleton; Golgi apparatus; Cytoplasm; Nucleus; Cytoplasmic vesicle, secretory vesicle, acrosome","url":"https://www.uniprot.org/uniprotkb/Q9Y615/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ACTL7A","classification":"Not Classified","n_dependent_lines":15,"n_total_lines":1208,"dependency_fraction":0.012417218543046357},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/ACTL7A","total_profiled":1310},"omim":[{"mim_id":"620500","title":"SPERMATOGENIC FAILURE 87; SPGF87","url":"https://www.omim.org/entry/620500"},{"mim_id":"620499","title":"SPERMATOGENIC FAILURE 86; SPGF86","url":"https://www.omim.org/entry/620499"},{"mim_id":"620170","title":"SPERMATOGENIC FAILURE 78; SPGF78","url":"https://www.omim.org/entry/620170"},{"mim_id":"620160","title":"IQ MOTIF-CONTAINING PROTEIN N; IQCN","url":"https://www.omim.org/entry/620160"},{"mim_id":"619251","title":"ACTIN-LIKE 9; ACTL9","url":"https://www.omim.org/entry/619251"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Perinuclear theca","reliability":"Supported"}],"tissue_specificity":"Tissue enriched","tissue_distribution":"Detected in single","driving_tissues":[{"tissue":"testis","ntpm":218.7}],"url":"https://www.proteinatlas.org/search/ACTL7A"},"hgnc":{"alias_symbol":[],"prev_symbol":[]},"alphafold":{"accession":"Q9Y615","domains":[{"cath_id":"3.30.420.40","chopping":"73-209_397-435","consensus_level":"medium","plddt":90.5615,"start":73,"end":435},{"cath_id":"3.90.640.10","chopping":"247-329","consensus_level":"high","plddt":94.5358,"start":247,"end":329}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y615","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y615-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y615-F1-predicted_aligned_error_v6.png","plddt_mean":84.56},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ACTL7A","jax_strain_url":"https://www.jax.org/strain/search?query=ACTL7A"},"sequence":{"accession":"Q9Y615","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9Y615.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9Y615/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y615"}},"corpus_meta":[{"pmid":"32923619","id":"PMC_32923619","title":"Disruption in ACTL7A causes acrosomal ultrastructural defects in human and mouse sperm as a novel male factor inducing early embryonic arrest.","date":"2020","source":"Science advances","url":"https://pubmed.ncbi.nlm.nih.gov/32923619","citation_count":80,"is_preprint":false},{"pmid":"34727571","id":"PMC_34727571","title":"Novel bi-allelic variants in ACTL7A are associated with male infertility and total fertilization failure.","date":"2021","source":"Human reproduction (Oxford, England)","url":"https://pubmed.ncbi.nlm.nih.gov/34727571","citation_count":34,"is_preprint":false},{"pmid":"10373328","id":"PMC_10373328","title":"Cloning, mapping, and expression of two novel actin genes, actin-like-7A (ACTL7A) and actin-like-7B (ACTL7B), from the familial dysautonomia candidate region on 9q31.","date":"1999","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/10373328","citation_count":30,"is_preprint":false},{"pmid":"36734600","id":"PMC_36734600","title":"Testis-specific actin-like 7A (ACTL7A) is an indispensable protein for subacrosomal-associated F-actin formation, acrosomal anchoring, and male fertility.","date":"2023","source":"Molecular human reproduction","url":"https://pubmed.ncbi.nlm.nih.gov/36734600","citation_count":22,"is_preprint":false},{"pmid":"35863052","id":"PMC_35863052","title":"Pathogenic variant in ACTL7A causes severe teratozoospermia characterized by bubble-shaped acrosomes and male infertility.","date":"2022","source":"Molecular human reproduction","url":"https://pubmed.ncbi.nlm.nih.gov/35863052","citation_count":21,"is_preprint":false},{"pmid":"36593593","id":"PMC_36593593","title":"Novel variants in ACTL7A and PLCZ1 are associated with male infertility and total fertilization failure.","date":"2023","source":"Clinical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/36593593","citation_count":17,"is_preprint":false},{"pmid":"22386842","id":"PMC_22386842","title":"Anti-ACTL7a antibodies: a cause of infertility.","date":"2012","source":"Fertility and 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andrology","url":"https://pubmed.ncbi.nlm.nih.gov/35532568","citation_count":11,"is_preprint":false},{"pmid":"37991128","id":"PMC_37991128","title":"Novel ACTL7A variants in males lead to fertilization failure and male infertility.","date":"2023","source":"Andrology","url":"https://pubmed.ncbi.nlm.nih.gov/37991128","citation_count":8,"is_preprint":false},{"pmid":"31310081","id":"PMC_31310081","title":"Genetic Polymorphisms within The Intronless ACTL7A and ACTL7B Genes Encoding Spermatogenesis-Specific Actin-Like Proteins in Japanese Males.","date":"2019","source":"International journal of fertility & sterility","url":"https://pubmed.ncbi.nlm.nih.gov/31310081","citation_count":6,"is_preprint":false},{"pmid":"38464253","id":"PMC_38464253","title":"Novel Nuclear Roles for Testis-Specific ACTL7A and ACTL7B Supported by In Vivo Characterizations and AI Facilitated In Silico Mechanistic Modeling with Implications for Epigenetic Regulation in Spermiogenesis.","date":"2024","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/38464253","citation_count":1,"is_preprint":false},{"pmid":"41169243","id":"PMC_41169243","title":"Perinuclear theca protein FNDC8 interacts with CCIN and ACTL7A to ensure proper sperm head shaping during spermiogenesis.","date":"2025","source":"Zoological research","url":"https://pubmed.ncbi.nlm.nih.gov/41169243","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.03.27.645694","title":"Actin-related protein M1 (ARPM1) required for acrosome biogenesis and sperm function in mice","date":"2025-04-01","source":"bioRxiv","url":"https://doi.org/10.1101/2025.03.27.645694","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11644,"output_tokens":3412,"usd":0.043056,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10951,"output_tokens":3827,"usd":0.075215,"stage2_stop_reason":"end_turn"},"total_usd":0.118271,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1999,\n      \"finding\": \"ACTL7A encodes a 435-amino-acid actin-like protein (predicted MW 48.6 kDa) expressed in multiple adult tissues, with the gene located intronless on chromosome 9q31 in a head-to-head orientation with ACTL7B on a common 8-kb HindIII fragment.\",\n      \"method\": \"cDNA selection, direct genomic sequencing, linkage analysis\",\n      \"journal\": \"Genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct cloning and genomic characterization with expression analysis across tissues, single lab, multiple methods\",\n      \"pmids\": [\"10373328\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"ACTL7A expression is upregulated via the PKA pathway and undergoes relocalization (remodeling) during the early period of capacitation in mouse spermatozoa, indicating it is an essential component of sperm capacitation.\",\n      \"method\": \"Western blot, indirect immunostaining during capacitation induction\",\n      \"journal\": \"Fertility and sterility\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — two methods (western blot + immunostaining), single lab, no genetic rescue or in vitro reconstitution\",\n      \"pmids\": [\"23211711\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Anti-ACTL7A antibodies cause sperm agglutination and reduce fertilizing capacity of mouse spermatozoa in vitro; active immunization of mice with ACTL7A protein significantly reduces fertility, establishing ACTL7A as a target antigen in immunologic infertility.\",\n      \"method\": \"In vitro sperm treatment with antibody-containing serum, active immunization fertility assay, sperm agglutination test, mass spectrometry identification\",\n      \"journal\": \"Fertility and sterility\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional fertility assay in vivo and in vitro, multiple orthogonal methods, single lab\",\n      \"pmids\": [\"22386842\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"A homozygous missense mutation in ACTL7A causes acrosomal ultrastructural defects in sperm and leads to reduced expression and abnormal localization of PLCζ in sperm, resulting in failure of oocyte activation and early embryonic arrest. Artificial oocyte activation rescues the fertilization defect.\",\n      \"method\": \"Whole-exome sequencing, knock-in mouse model, transmission electron microscopy, immunofluorescence, western blot\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic patient data corroborated by knock-in mouse model with consistent phenotype, multiple orthogonal methods (TEM, IF, WB), functional rescue by AOA\",\n      \"pmids\": [\"32923619\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Compound heterozygous loss-of-function variants in ACTL7A cause ultrastructural defects in the acrosome and perinuclear theca, and significantly reduce expression of both ACTL7A protein and PLCζ in sperm, leading to oocyte activation deficiency and total fertilization failure.\",\n      \"method\": \"Whole-exome sequencing, Sanger sequencing, transmission electron microscopy, immunofluorescence, western blot\",\n      \"journal\": \"Human reproduction\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods, single family/lab, replicates mechanism from prior study\",\n      \"pmids\": [\"34727571\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"In Actl7a knockout mice, sperm show malformed acrosomes, altered localization of zona pellucida binding protein ZPBP, and reduced calcium oscillations in oocytes due to abnormal localization and expression of PLCZ1; ACTL7A and ZPBP co-immunoprecipitate, forming a complex potentially involved in acrosomal formation.\",\n      \"method\": \"Knockout mouse model, immunofluorescence, Co-immunoprecipitation, western blot, IVF/ICSI\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO mouse with defined phenotype plus Co-IP for complex formation, single lab\",\n      \"pmids\": [\"35921706\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"A pathogenic variant in ACTL7A (p.Gly402Ser) causes mutant ACTL7A to fail to attach to the acroplaxome and be discharged via cytoplasmic droplets, resulting in absence of ACTL7A from epididymal sperm, acrosome detachment from the nuclear membrane (bubble-shaped acrosomes), and PLCζ co-discharge leading to total fertilization failure. Immunoprecipitation-mass spectrometry identified interacting proteins involved in acrosome assembly and actin filament organization.\",\n      \"method\": \"Knock-in mouse model (equivalent variant), transmission electron microscopy, immunofluorescence, immunoprecipitation/LC-MS, western blot\",\n      \"journal\": \"Molecular human reproduction\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — knock-in mouse model replicating patient phenotype, multiple orthogonal methods including IP-MS for interactome, mechanistic explanation of protein mislocalization and discharge\",\n      \"pmids\": [\"35863052\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"A homozygous missense mutation p.D75A in ACTL7A causes protein degradation in sperm, irregular perinuclear theca and acrosomal ultrastructural defects, and abnormal localization and reduced expression of PLCZ1; 3D structural modeling shows loss of a hydrogen bond with Ser170 and transformation of an α-helix to random coil.\",\n      \"method\": \"Whole-exome sequencing, 3D structural modeling, immunofluorescence, transmission electron microscopy, western blot\",\n      \"journal\": \"Molecular genetics and genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — multiple clinical and experimental methods, structural modeling is in silico only, single lab\",\n      \"pmids\": [\"36574082\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ACTL7A is dynamically localized within the nucleus and subacrosomal space of developing spermatids, and later associates with postacrosomal regions. Actl7a knockout mice show complete loss of subacrosomal filamentous actin (F-actin) structures, abnormal acrosomal granule migration, and peeling acrosomes during spermatid elongation, establishing ACTL7A as required for subacrosomal F-actin formation and acrosomal anchoring via the acroplaxome.\",\n      \"method\": \"Actl7a knockout mouse model, immunofluorescence (dynamic localization), phalloidin staining for F-actin, transmission electron microscopy\",\n      \"journal\": \"Molecular human reproduction\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — KO mouse model with multiple orthogonal methods, direct demonstration of F-actin loss as mechanistic link, single lab with rigorous controls\",\n      \"pmids\": [\"36734600\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Loss of ACTL7A disrupts the acrosome-acroplaxome-manchette complex, causing small head sperm. Proteomic analysis of Actl7a-KO testes reveals enrichment of differentially expressed proteins in the PI3K/AKT/mTOR pathway; autophagy inhibition via PI3K/AKT/mTOR activation leads to PDLIM1 accumulation, impairing manchette development and sperm head shaping.\",\n      \"method\": \"Actl7a knockout mouse model, immunofluorescence, transmission electron microscopy, tandem mass tag quantitative proteomics, western blot\",\n      \"journal\": \"Reproductive biology and endocrinology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO mouse with proteomics and multiple validation methods, PI3K/AKT/mTOR/autophagy pathway placement, single lab\",\n      \"pmids\": [\"37667331\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ACTL7A variants affecting the actin domain cause absent ACTL7A protein in spermatozoa and near-absent PLCζ1, along with attenuated and unevenly distributed acrosomal PNA signals, indicating acrosome dysfunction and oocyte activation failure.\",\n      \"method\": \"Whole-exome sequencing, immunofluorescence, western blot, bioinformatic structural prediction\",\n      \"journal\": \"Andrology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — multiple patient cases with consistent findings, immunofluorescence and WB, no KO model, single lab\",\n      \"pmids\": [\"37991128\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In the absence of ACTL7A (or ACTL7B), intranuclear localization of HDAC1 and HDAC3 is lost in spermatids, implicating ACTL7A in nuclear HDAC association and epigenetic regulation during spermiogenesis. In silico modeling predicts ACTL7A can bind to HSA domains of INO80 and SWI/SNF nucleosome remodeler family members in a manner analogous to nuclear actin and ACTL6A, suggesting ARP subunit swapping in chromatin regulatory complexes.\",\n      \"method\": \"Actl7a and Actl7b knockout mouse models, immunofluorescence for HDAC1/HDAC3 localization, transcriptomic analysis, in silico structural modeling\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — preprint, HDAC mislocalization shown by IF in KO mice (moderate evidence) but nucleosome remodeler interactions are in silico predictions only, not biochemically validated\",\n      \"pmids\": [\"38464253\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"FNDC8 interacts with ACTL7A (and CCIN) in the perinuclear theca during spermiogenesis; depletion of FNDC8 destabilizes ACTL7A protein levels, establishing ACTL7A as part of a protein complex within the perinuclear theca that maintains structural integrity for sperm head morphogenesis.\",\n      \"method\": \"Co-immunoprecipitation, Fndc8 knockout mouse model, immunofluorescence, western blot\",\n      \"journal\": \"Zoological research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP demonstrating interaction, KO mouse showing destabilization of ACTL7A, single lab\",\n      \"pmids\": [\"41169243\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ACTL7A co-immunoprecipitates with the perinuclear theca proteins ACTRT1, ACTRT2, and ARPM1, and with the sperm surface protein ZPBP, placing ACTL7A within a cytoskeletal network in the perinuclear theca that connects the acrosome and nucleus.\",\n      \"method\": \"Co-immunoprecipitation in Arpm1-deficient mouse model context\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — preprint, single Co-IP experiment, ACTL7A is not the primary subject of the study\",\n      \"pmids\": [\"bio_10.1101_2025.03.27.645694\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"ACTL7A is a testis-enriched actin-like protein that localizes dynamically to the nucleus, subacrosomal space, and postacrosomal regions of developing spermatids, where it is required for subacrosomal F-actin formation, acroplaxome-mediated acrosomal attachment to the nucleus, and sperm head shaping; loss-of-function mutations cause acrosome detachment, perinuclear theca defects, and mislocalization/loss of PLCζ (PLCZ1), the sperm-borne oocyte activation factor, resulting in oocyte activation failure, total fertilization failure, and male infertility that can be rescued by artificial oocyte activation.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ACTL7A is a testis-enriched actin-like protein essential for acrosome biogenesis and sperm head shaping during spermiogenesis [#8]. It localizes dynamically to the nucleus and subacrosomal space of developing spermatids and later to postacrosomal regions, where it drives formation of subacrosomal filamentous actin and anchors the acrosome to the nucleus via the acroplaxome; its loss abolishes subacrosomal F-actin, causes abnormal acrosomal granule migration, and produces peeling, detached acrosomes [#8]. ACTL7A operates within a perinuclear theca cytoskeletal network, co-immunoprecipitating with the zona-pellucida-binding protein ZPBP and being stabilized by FNDC8, whose depletion destabilizes ACTL7A and disrupts head morphogenesis [#5, #12]. Disruption of the acrosome-acroplaxome-manchette complex upon ACTL7A loss yields small-headed sperm, accompanied by dysregulation of the PI3K/AKT/mTOR/autophagy axis and PDLIM1 accumulation that impairs manchette development [#9]. A central downstream consequence of ACTL7A dysfunction is mislocalization, reduced expression, or co-discharge of the sperm-borne oocyte activation factor PLCζ (PLCZ1), which abolishes oocyte calcium oscillations and causes total fertilization failure [#3, #6]. Loss-of-function and missense mutations in ACTL7A cause male infertility through acrosomal and perinuclear theca ultrastructural defects and oocyte activation deficiency, a phenotype rescuable by artificial oocyte activation [#3, #4].\",\n  \"teleology\": [\n    {\n      \"year\": 1999,\n      \"claim\": \"Established ACTL7A's molecular identity by cloning it as an intronless actin-like gene paired head-to-head with ACTL7B, defining the basic gene and protein before any functional role was known.\",\n      \"evidence\": \"cDNA selection and direct genomic sequencing with multi-tissue expression analysis\",\n      \"pmids\": [\"10373328\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No protein function or subcellular role defined\", \"Testis-specific role not yet identified\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"First linked ACTL7A to sperm physiology by showing PKA-driven upregulation and relocalization during capacitation, and demonstrating that antibody targeting impairs fertilization, framing it as a functionally important and immunologically accessible sperm antigen.\",\n      \"evidence\": \"Western blot and immunostaining during capacitation; in vitro sperm antibody treatment and active immunization fertility assays in mouse\",\n      \"pmids\": [\"23211711\", \"22386842\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism of relocalization unknown\", \"No genetic loss-of-function evidence\", \"Direct binding partners not identified\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Connected ACTL7A mutation to human infertility by showing a missense variant causes acrosomal defects and PLCζ mislocalization that block oocyte activation, with rescue by artificial oocyte activation establishing the therapeutic and mechanistic link.\",\n      \"evidence\": \"Whole-exome sequencing, knock-in mouse, TEM, immunofluorescence, western blot, AOA rescue\",\n      \"pmids\": [\"32923619\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How ACTL7A controls PLCζ localization not resolved\", \"Structural basis of the missense effect not defined\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Confirmed and broadened the disease link by showing biallelic loss-of-function variants reduce ACTL7A and PLCζ and produce perinuclear theca defects, reinforcing oocyte activation deficiency as the core pathology.\",\n      \"evidence\": \"Whole-exome and Sanger sequencing, TEM, immunofluorescence, western blot in patient sperm\",\n      \"pmids\": [\"34727571\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No mouse model in this study\", \"Mechanistic coupling of PT integrity to PLCζ retention unclear\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined ACTL7A's physical partnerships and the consequence of mislocalization, showing it complexes with ZPBP and that variant protein fails to attach to the acroplaxome and is discharged with PLCζ, explaining the loss of both proteins from sperm.\",\n      \"evidence\": \"Knockout and knock-in mouse models, Co-IP, IP-LC/MS interactome, TEM, immunofluorescence, IVF/ICSI\",\n      \"pmids\": [\"35921706\", \"35863052\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry and direct vs indirect nature of interactions not resolved\", \"How acroplaxome attachment is molecularly mediated unknown\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Provided structural rationale for a pathogenic missense allele, showing p.D75A destabilizes ACTL7A via loss of a hydrogen bond and helix-to-coil transition, linking protein folding to PT/acrosome defects and PLCZ1 dysregulation.\",\n      \"evidence\": \"Whole-exome sequencing, in silico 3D structural modeling, immunofluorescence, TEM, western blot\",\n      \"pmids\": [\"36574082\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural prediction not experimentally validated\", \"Degradation pathway not identified\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identified the core cell-biological function as subacrosomal F-actin formation and acroplaxome-mediated acrosomal anchoring, the mechanistic basis for acrosome attachment that earlier mutation studies implicated indirectly.\",\n      \"evidence\": \"Actl7a knockout mouse, phalloidin F-actin staining, dynamic-localization immunofluorescence, TEM\",\n      \"pmids\": [\"36734600\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Biochemical mechanism of F-actin nucleation/organization by ACTL7A unknown\", \"Whether ACTL7A polymerizes or scaffolds conventional actin unresolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Placed ACTL7A within a broader head-shaping program, linking its loss to acrosome-acroplaxome-manchette disruption and to PI3K/AKT/mTOR-mediated autophagy inhibition and PDLIM1 accumulation that impairs the manchette.\",\n      \"evidence\": \"Actl7a knockout mouse, TMT quantitative proteomics, immunofluorescence, TEM, western blot\",\n      \"pmids\": [\"37667331\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Causality between ACTL7A loss and pathway activation not directly tested\", \"Direct vs secondary effect on manchette unclear\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended the human genetic spectrum to actin-domain variants, confirming absent sperm ACTL7A, near-absent PLCζ, and impaired acrosomal PNA staining as a reproducible patient phenotype.\",\n      \"evidence\": \"Whole-exome sequencing, immunofluorescence, western blot, bioinformatic structural prediction in patients\",\n      \"pmids\": [\"37991128\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No functional model in this cohort\", \"Genotype-phenotype correlation across variants incomplete\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined a perinuclear theca complex by showing FNDC8 interacts with and stabilizes ACTL7A, providing a mechanism for maintaining ACTL7A protein levels needed for head morphogenesis.\",\n      \"evidence\": \"Co-immunoprecipitation and Fndc8 knockout mouse, immunofluorescence, western blot\",\n      \"pmids\": [\"41169243\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether stabilization is direct or via complex assembly unknown\", \"Other PT partners maintaining ACTL7A not fully mapped\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Began mapping a wider PT cytoskeletal network by Co-IP of ACTL7A with ACTRT1/ACTRT2/ARPM1 and ZPBP, and proposed a nuclear chromatin-regulatory role via HDAC1/HDAC3 association and putative ARP subunit swapping in remodeler complexes.\",\n      \"evidence\": \"Co-IP in Arpm1-deficient context (preprint); Actl7a/Actl7b knockout mice with HDAC1/HDAC3 immunofluorescence, transcriptomics, in silico modeling (preprint)\",\n      \"pmids\": [\"bio_10.1101_2025.03.27.645694\", \"38464253\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single Co-IP without reciprocal validation; ACTL7A not the primary subject\", \"Nucleosome-remodeler interactions are in silico predictions only, not biochemically validated\", \"Nuclear/chromatin function not connected to the cytoplasmic acrosomal role\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The biochemical mechanism by which ACTL7A nucleates or organizes subacrosomal F-actin, and how this physically retains PLCζ in the perinuclear theca, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No in vitro reconstitution of ACTL7A actin activity\", \"Direct ACTL7A-PLCζ interaction not demonstrated\", \"Structural model of ACTL7A in the acroplaxome lacking\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [8]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [8, 13]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [8, 11]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [8, 13]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1474165\", \"supporting_discovery_ids\": [3, 8]}\n    ],\n    \"complexes\": [\n      \"acroplaxome\",\n      \"perinuclear theca cytoskeletal network\",\n      \"acrosome-acroplaxome-manchette complex\"\n    ],\n    \"partners\": [\n      \"ZPBP\",\n      \"FNDC8\",\n      \"CCIN\",\n      \"ACTRT1\",\n      \"ACTRT2\",\n      \"ARPM1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":6,"faith_pct":83.33333333333333}}