{"gene":"WAC","run_date":"2026-04-28T23:00:23","timeline":{"discoveries":[{"year":2011,"finding":"WAC was identified as a functional partner of RNF20/40 through protein affinity purification. WAC interacts with RNF20/40 via its C-terminal coiled-coil region and promotes RNF20/40's E3 ligase activity for histone H2B ubiquitination. The N-terminal WW domain of WAC recognizes RNA polymerase II, enabling WAC to target RNF20/40 to associate with the RNA polymerase II complex at active transcription sites.","method":"Protein affinity purification, Co-IP, domain mutagenesis, siRNA knockdown, H2B ubiquitination assay","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1-2 — multiple orthogonal methods (affinity purification, domain mutagenesis, functional ubiquitination assay) in a highly-cited foundational paper","pmids":["21329877"],"is_preprint":false},{"year":2011,"finding":"WAC-dependent H2B ubiquitination and transcription regulation is important for cell-cycle checkpoint activation in response to genotoxic stress, as depletion of WAC abolished H2B ubiquitination and compromised checkpoint activation.","method":"siRNA knockdown, genotoxic stress assays, H2B ubiquitination assay","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 — clean knockdown with defined cellular phenotype, supported by multiple assays in a highly-cited paper","pmids":["21329877"],"is_preprint":false},{"year":2011,"finding":"WAC localizes to the Golgi and nucleus and was identified as a VCIP135-binding protein. WAC directly binds VCIP135 and increases its deubiquitinating activity, and WAC is required for p97/p47-mediated Golgi membrane fusion (but not p97/p37-mediated reassembly or ER membrane fusion).","method":"Co-IP, siRNA knockdown, in vitro Golgi reformation assay, deubiquitinase activity assay","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1-2 — direct binding shown, enzymatic activity assay performed, in vitro reconstitution of Golgi reformation, clean siRNA knockdown with specific pathway phenotype","pmids":["21811234"],"is_preprint":false},{"year":2012,"finding":"WAC is required for starvation-induced autophagosome formation, identified in a genome-wide siRNA screen; WAC also acts as a potential negative regulator of the ubiquitin-proteasome system.","method":"Genome-wide siRNA screen, GFP-LC3 autophagosome marker assay, siRNA knockdown validation","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 — genome-wide screen with stringent validation criteria, multiple orthogonal validation steps, replicated in follow-up studies","pmids":["22354037"],"is_preprint":false},{"year":2015,"finding":"WAC is bound to the Golgi by GM130 (a direct interaction required for autophagy). WAC and GM130 interact with the Atg8 homolog GABARAP and regulate its subcellular localization: GABARAP resides on the pericentriolar matrix, and WAC suppresses GM130 binding to GABARAP, enabling starvation-induced delivery of centrosomal GABARAP to the phagophore to activate ULK kinase.","method":"Co-IP, pulldown, siRNA knockdown, live-cell imaging, subcellular fractionation, ULK kinase activation assay","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1-2 — direct interaction established, localization studies with functional consequence, multiple orthogonal methods in a highly-cited paper","pmids":["26687599"],"is_preprint":false},{"year":2002,"finding":"WAC was identified as a novel WW domain-containing adaptor protein that colocalizes with splicing factor SC35 (a marker for pre-mRNA splicing machinery) and exists mainly in a tyrosine-phosphorylated form in cells.","method":"Immunofluorescence, domain analysis, phosphorylation analysis","journal":"Genomics","confidence":"Medium","confidence_rationale":"Tier 3 — localization by immunofluorescence without deep functional follow-up; single method for colocalization","pmids":["11827461"],"is_preprint":false},{"year":2018,"finding":"WAC promotes Polo-like kinase 1 (Plk1) activation for timely mitotic entry: Cdk1 phosphorylates WAC, priming its direct interaction with the polo-box domain of Plk1. WAC also binds Aurora A kinase (AurkA) and can enhance Plk1 phosphorylation by AurkA in vitro. Knockdown of WAC compromises Plk1 activity and delays mitotic entry, rescued by wild-type but not Plk1-binding-deficient WAC.","method":"Co-IP, in vitro kinase assay, mutagenesis, siRNA knockdown with rescue experiment, mitotic entry assay","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 1-2 — in vitro kinase assay, direct interaction established, mutagenesis rescue experiment, clean loss-of-function phenotype","pmids":["30021153"],"is_preprint":false},{"year":2017,"finding":"WAC functions as part of the RNF20/RNF40/WAC E3 ligase complex for H2B ubiquitination in leukemia cells; knockdown of WAC phenocopied loss of H2B ubiquitination and induced cell death in MLL-rearranged ALL.","method":"siRNA knockdown, H2B ubiquitination assay, cell viability assay","journal":"Leukemia","confidence":"Medium","confidence_rationale":"Tier 2 — clean knockdown with specific molecular and cellular phenotype, consistent with foundational mechanistic work","pmids":["28690313"],"is_preprint":false},{"year":2023,"finding":"B cell-specific Wac knockout mice show severely compromised T cell-dependent and -independent antibody responses, with drastically reduced plasma cell differentiation but intact germinal center B cell responses, linked to significant reduction in global H2B ubiquitination (ubH2B) in Wac-deficient B cells.","method":"Conditional B cell-specific knockout mouse model, antibody response assays, ubH2B western blotting, flow cytometry","journal":"FEBS letters","confidence":"High","confidence_rationale":"Tier 2 — clean conditional KO with defined cellular phenotype and molecular mechanism (ubH2B reduction)","pmids":["37171241"],"is_preprint":false},{"year":2024,"finding":"WAC interacts with the transmembrane (TM) domains of PINK1 and prevents ubiquitination at the K137 site of PINK1, protecting PINK1 from ubiquitination-dependent degradation and thereby activating mitophagy to promote MSC osteogenesis.","method":"Co-IP, ubiquitination assay, mutagenesis (K137 site), in vitro and in vivo osteogenesis assays","journal":"Advanced science","confidence":"Medium","confidence_rationale":"Tier 2 — direct interaction and site-specific ubiquitination shown, functional rescue in osteogenesis; single lab","pmids":["39555688"],"is_preprint":false},{"year":2025,"finding":"WAC directly binds mTOR-mLST8, R2TP, and TELO2 (but not TTI1 and TTI2) using purified proteins. In cells, WAC is part of complexes containing mTORC1, R2TP, and TTT components, and WAC and TELO2 strongly associate with mTOR under glucose and glutamine deprivation, with these interactions weakened upon nutrient refeeding, correlating with mTORC1 activity changes.","method":"Purified protein binding assays, Co-IP in cells under nutrient stress, transcriptomics, proteomics","journal":"FEBS open bio","confidence":"Medium","confidence_rationale":"Tier 1-2 — direct binding established with purified proteins, in-cell nutrient-responsive dynamics measured; single study","pmids":["40653822"],"is_preprint":false},{"year":2026,"finding":"Crystal structure of the yeast Bre1-Lge1 complex and AlphaFold-predicted structure of human RNF20/RNF40-WAC, combined with in vitro and in vivo experiments, revealed extensive interaction interfaces and identified key electrostatic interactions critical for binding specificity and H2B ubiquitination activity.","method":"X-ray crystallography (Bre1-Lge1), AlphaFold structure prediction (RNF20/RNF40-WAC), mutagenesis, in vitro ubiquitination assay, in vivo functional assays","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 1 — crystal structure of orthologous complex, structure-guided mutagenesis validated in vitro and in vivo","pmids":["41533567"],"is_preprint":false},{"year":2025,"finding":"WAC regulates H2BK120ub1 and influences H3K27me3 levels in chondrocytes by regulating nuclear entry of the H3K27 demethylase KDM6B, acting as a key crosstalk factor between H2BK120ub1 and H3K27me3 to control inflammatory mediator secretion and cartilage degradation.","method":"Cartilage-specific WAC knockout mice, H2BK120ub1 and H3K27me3 ChIP/western blotting, KDM6B nuclear entry assays, CIA and CIOA mouse models","journal":"Acta pharmaceutica sinica. B","confidence":"Medium","confidence_rationale":"Tier 2 — conditional KO with defined molecular phenotype (crosstalk between histone marks), in vivo validation; single lab","pmids":["40893665"],"is_preprint":false},{"year":2023,"finding":"WAC protein is expressed in a developmental stage-dependent manner in mouse brain and localizes predominantly to the perinuclear region of cortical neurons embryonically, then becomes enriched in the nucleus after birth; WAC is also present in axons and dendrites of cultured hippocampal neurons in a time-dependent manner.","method":"Anti-WAC antibody generation, western blotting, immunohistochemistry, immunofluorescence of primary cultured neurons","journal":"Medical molecular morphology","confidence":"Medium","confidence_rationale":"Tier 2-3 — direct localization with developmental context, but functional consequence not directly demonstrated in this study","pmids":["37402055"],"is_preprint":false},{"year":2023,"finding":"A nuclear localization domain in WAC impacts the cellular distribution of the protein in cortical GABAergic neurons, as shown by deletion analysis of human protein domains.","method":"Domain deletion constructs, cellular distribution assay in GABAergic neurons","journal":"Biology","confidence":"Medium","confidence_rationale":"Tier 2-3 — functional domain deletion with localization readout; single study","pmids":["37106788"],"is_preprint":false}],"current_model":"WAC is a WW domain- and coiled-coil-containing adaptor protein that promotes histone H2B ubiquitination by binding RNF20/40 (via its coiled-coil) and recruiting this E3 ligase complex to RNA polymerase II at active transcription sites (via its WW domain); it also regulates starvation-induced autophagy by modulating GABARAP localization between the Golgi (where GM130 sequesters it) and the centrosome/phagophore to activate ULK kinase, activates Plk1 for mitotic entry after Cdk1-mediated phosphorylation, protects PINK1 from ubiquitination-dependent degradation to facilitate mitophagy, and interacts with mTOR-mLST8, R2TP, and TELO2 to regulate mTORC1 activity in response to nutrient availability."},"narrative":{"teleology":[{"year":2002,"claim":"Initial identification of WAC as a novel WW domain-containing adaptor established its domain architecture and suggested a connection to nuclear RNA processing through colocalization with splicing factor SC35.","evidence":"Immunofluorescence and domain analysis in cultured cells","pmids":["11827461"],"confidence":"Medium","gaps":["Colocalization with SC35 shown by single method without functional consequence demonstrated","No interacting partners identified","Tyrosine phosphorylation significance unknown"]},{"year":2011,"claim":"The discovery that WAC bridges RNF20/RNF40 to RNA polymerase II via distinct domains to promote H2B ubiquitination established WAC's primary molecular function as a transcription-coupled chromatin modification scaffold, and linked this activity to genotoxic stress checkpoint activation.","evidence":"Affinity purification, Co-IP, domain mutagenesis, siRNA knockdown, H2B ubiquitination and checkpoint assays in human cells","pmids":["21329877"],"confidence":"High","gaps":["Structural basis of WAC-RNF20/40 interaction not resolved at atomic level","In vivo physiological relevance in organism not tested"]},{"year":2011,"claim":"Parallel work revealed a second function for WAC at the Golgi: direct binding and activation of the deubiquitinase VCIP135 for p97/p47-dependent Golgi membrane fusion, establishing WAC as a bifunctional nuclear-Golgi protein.","evidence":"Co-IP, in vitro Golgi reformation assay, DUB activity assay, siRNA knockdown","pmids":["21811234"],"confidence":"High","gaps":["How WAC partitions between nuclear and Golgi functions is unclear","Ubiquitin substrates of VCIP135 at the Golgi not identified"]},{"year":2012,"claim":"A genome-wide siRNA screen identified WAC as required for starvation-induced autophagosome formation, opening a third functional axis beyond chromatin and Golgi membrane dynamics.","evidence":"Genome-wide siRNA screen with GFP-LC3 reporter validation","pmids":["22354037"],"confidence":"High","gaps":["Molecular mechanism connecting WAC to autophagy not yet defined","Potential indirect effects through H2B ubiquitination not excluded"]},{"year":2015,"claim":"The autophagy mechanism was resolved: WAC is tethered to the Golgi by GM130 and competes with GM130 for GABARAP binding, enabling starvation-triggered release of centrosomal GABARAP to phagophores where it activates ULK kinase.","evidence":"Co-IP, pulldown, live-cell imaging, subcellular fractionation, ULK kinase activation assay","pmids":["26687599"],"confidence":"High","gaps":["Signal that switches WAC from GM130-bound to GABARAP-releasing state not identified","Whether WAC regulation of GABARAP is tissue-specific is unknown"]},{"year":2018,"claim":"WAC was shown to promote mitotic entry by acting as a Cdk1-phosphorylated scaffold that recruits Plk1 via its polo-box domain and enhances Plk1 phosphorylation by Aurora A, revealing a cell-cycle regulatory function.","evidence":"In vitro kinase assays, Co-IP, mutagenesis with rescue experiments, mitotic entry assays","pmids":["30021153"],"confidence":"High","gaps":["Whether WAC's mitotic role is coordinated with its chromatin or autophagy functions is unknown","Structural details of the WAC-Plk1-AurkA ternary complex not resolved"]},{"year":2023,"claim":"Conditional B cell-specific Wac knockout demonstrated that WAC-dependent H2B ubiquitination is essential in vivo for plasma cell differentiation and antibody responses, while germinal center formation is spared.","evidence":"Conditional knockout mouse model, flow cytometry, antibody response assays, ubH2B western blotting","pmids":["37171241"],"confidence":"High","gaps":["Transcriptional targets downstream of WAC-dependent ubH2B in plasma cells not identified","Whether other H2B ubiquitination-independent WAC functions contribute to the B cell phenotype is untested"]},{"year":2023,"claim":"Characterization of WAC protein expression in developing mouse brain showed stage-dependent localization from perinuclear (embryonic) to nuclear (postnatal) in cortical neurons, with a nuclear localization domain identified by deletion analysis.","evidence":"Immunohistochemistry, immunofluorescence, domain deletion constructs in neurons","pmids":["37402055","37106788"],"confidence":"Medium","gaps":["Functional consequence of developmental relocalization not demonstrated","Domain deletion studies performed only in GABAergic neurons","No loss-of-function neuronal phenotype directly shown"]},{"year":2024,"claim":"WAC was found to protect PINK1 from ubiquitination-dependent degradation by binding its transmembrane domain and blocking K137 ubiquitination, thereby activating mitophagy to promote MSC osteogenesis.","evidence":"Co-IP, ubiquitination assay with K137 mutagenesis, in vitro and in vivo osteogenesis assays","pmids":["39555688"],"confidence":"Medium","gaps":["Single-lab finding not independently replicated","Whether WAC-PINK1 interaction occurs broadly or is specific to MSC context is unclear","E3 ligase targeting K137 not identified"]},{"year":2025,"claim":"Direct binding assays with purified proteins established that WAC interacts with mTOR-mLST8, R2TP, and TELO2 in a nutrient-responsive manner, with associations strengthened during glucose/glutamine deprivation and weakened upon refeeding, linking WAC to mTORC1 regulation.","evidence":"Purified protein binding assays, Co-IP under nutrient stress, proteomics","pmids":["40653822"],"confidence":"Medium","gaps":["Single study; independent replication needed","Functional consequence of WAC on mTORC1 signaling output not directly demonstrated","Relationship to WAC's autophagy function via GABARAP/ULK pathway not resolved"]},{"year":2025,"claim":"Cartilage-specific WAC knockout revealed that WAC-dependent H2BK120ub1 regulates H3K27me3 levels by controlling nuclear entry of the demethylase KDM6B, establishing WAC as a histone mark crosstalk regulator with consequences for inflammatory cartilage degradation.","evidence":"Conditional knockout mice, ChIP, western blotting, KDM6B nuclear entry assays, arthritis mouse models","pmids":["40893665"],"confidence":"Medium","gaps":["Mechanism by which H2BK120ub1 controls KDM6B nuclear entry not molecularly defined","Single-lab finding","Whether this crosstalk operates in other tissues unknown"]},{"year":2026,"claim":"Crystal structure of the yeast Bre1-Lge1 complex and AlphaFold-predicted structure of human RNF20/RNF40-WAC resolved the interaction interfaces and identified electrostatic contacts critical for binding specificity and H2B ubiquitination, providing the first structural framework for this complex.","evidence":"X-ray crystallography of yeast orthologs, AlphaFold prediction for human complex, structure-guided mutagenesis with in vitro and in vivo validation","pmids":["41533567"],"confidence":"High","gaps":["Human complex structure predicted rather than experimentally determined","No structure of WAC WW domain bound to RNA Pol II CTD","Structural basis of WAC's non-chromatin functions unexplored"]},{"year":null,"claim":"How WAC coordinates its multiple scaffold functions—chromatin modification, autophagy, mitotic entry, Golgi membrane fusion, mTORC1 signaling, and mitophagy—in a cell-state-dependent manner remains an open question, as does the upstream regulatory logic that partitions WAC among these pathways.","evidence":"","pmids":[],"confidence":"Low","gaps":["No integrative study examining how WAC's multiple functions are coordinated or mutually exclusive","Full post-translational modification map of WAC not established","No disease-causative mutations in WAC identified through human genetic studies in the timeline"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,4,6,10]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[2,9]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,5,13,14]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[2,4]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[6,10]}],"pathway":[{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[0,1,7,8,11,12]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[3,4]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[6]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[0,1,12]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[10]}],"complexes":["RNF20/RNF40/WAC E3 ligase complex","mTORC1-R2TP-TTT complex"],"partners":["RNF20","RNF40","GM130","GABARAP","PLK1","VCIP135","PINK1","TELO2"],"other_free_text":[]},"mechanistic_narrative":"WAC is a WW domain- and coiled-coil-containing adaptor protein that functions as a central scaffold linking chromatin modification, autophagy, cell-cycle progression, and nutrient signaling. Its best-characterized role is promoting histone H2B monoubiquitination by binding the RNF20/RNF40 E3 ligase complex through its coiled-coil domain and recruiting it to RNA polymerase II at active transcription sites via its WW domain, a function critical for transcription-coupled checkpoint activation, plasma cell differentiation, and histone mark crosstalk with H3K27me3 through regulation of KDM6B nuclear entry [PMID:21329877, PMID:37171241, PMID:40893665, PMID:41533567]. WAC also regulates starvation-induced autophagy by competing with GM130 for GABARAP binding at the Golgi, thereby enabling delivery of centrosomal GABARAP to the phagophore to activate ULK kinase, and separately promotes mitophagy by protecting PINK1 from ubiquitination-dependent degradation [PMID:26687599, PMID:39555688]. Additionally, WAC facilitates timely mitotic entry by serving as a Cdk1-phosphorylated scaffold that bridges Aurora A and Plk1, participates in p97/p47-mediated Golgi membrane fusion by binding and activating the deubiquitinase VCIP135, and associates with mTOR-mLST8/R2TP/TELO2 complexes in a nutrient-responsive manner to modulate mTORC1 activity [PMID:30021153, PMID:21811234, PMID:40653822]."},"prefetch_data":{"uniprot":{"accession":"Q9BTA9","full_name":"WW domain-containing adapter protein with coiled-coil","aliases":[],"length_aa":647,"mass_kda":70.7,"function":"Acts as a linker between gene transcription and histone H2B monoubiquitination at 'Lys-120' (H2BK120ub1) (PubMed:21329877). Interacts with the RNA polymerase II transcriptional machinery via its WW domain and with RNF20-RNF40 via its coiled coil region, thereby linking and regulating H2BK120ub1 and gene transcription (PubMed:21329877). Regulates the cell-cycle checkpoint activation in response to DNA damage (PubMed:21329877). Positive regulator of amino acid starvation-induced autophagy (PubMed:22354037). Also acts as a negative regulator of basal autophagy (PubMed:26812014). Positively regulates MTOR activity by promoting, in an energy-dependent manner, the assembly of the TTT complex composed of TELO2, TTI1 and TTI2 and the RUVBL complex composed of RUVBL1 and RUVBL2 into the TTT-RUVBL complex. This leads to the dimerization of the mTORC1 complex and its subsequent activation (PubMed:26812014). May negatively regulate the ubiquitin proteasome pathway (PubMed:21329877)","subcellular_location":"Nucleus speckle; Nucleus","url":"https://www.uniprot.org/uniprotkb/Q9BTA9/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/WAC","classification":"Not Classified","n_dependent_lines":474,"n_total_lines":1208,"dependency_fraction":0.3923841059602649},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"RNF40","stoichiometry":10.0},{"gene":"RBM12","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/WAC","total_profiled":1310},"omim":[{"mim_id":"616708","title":"DESANTO-SHINAWI SYNDROME; DESSH","url":"https://www.omim.org/entry/616708"},{"mim_id":"615049","title":"WW DOMAIN-CONTAINING ADAPTOR WITH COILED-COIL REGION; WAC","url":"https://www.omim.org/entry/615049"},{"mim_id":"605681","title":"BROMODOMAIN ADJACENT TO ZINC FINGER DOMAIN, 1B; BAZ1B","url":"https://www.omim.org/entry/605681"},{"mim_id":"605680","title":"BROMODOMAIN ADJACENT TO ZINC FINGER DOMAIN, 1A; BAZ1A","url":"https://www.omim.org/entry/605680"},{"mim_id":"602468","title":"PROTEIN PHOSPHATASE 1, REGULATORY SUBUNIT 9A; PPP1R9A","url":"https://www.omim.org/entry/602468"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Nucleoplasm","reliability":"Enhanced"},{"location":"Centrosome","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/WAC"},"hgnc":{"alias_symbol":["Wwp4","FLJ31290","PRO1741","BM-016","MGC10753"],"prev_symbol":[]},"alphafold":{"accession":"Q9BTA9","domains":[{"cath_id":"2.20.70.10","chopping":"134-179","consensus_level":"medium","plddt":77.9983,"start":134,"end":179},{"cath_id":"-","chopping":"364-415","consensus_level":"high","plddt":68.6919,"start":364,"end":415}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BTA9","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BTA9-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BTA9-F1-predicted_aligned_error_v6.png","plddt_mean":54.84},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=WAC","jax_strain_url":"https://www.jax.org/strain/search?query=WAC"},"sequence":{"accession":"Q9BTA9","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9BTA9.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9BTA9/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BTA9"}},"corpus_meta":[{"pmid":"21329877","id":"PMC_21329877","title":"WAC, a functional partner of RNF20/40, regulates histone H2B ubiquitination and gene transcription.","date":"2011","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/21329877","citation_count":132,"is_preprint":false},{"pmid":"26687599","id":"PMC_26687599","title":"Activation of ULK Kinase and Autophagy by GABARAP Trafficking from the Centrosome Is Regulated by WAC and GM130.","date":"2015","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/26687599","citation_count":117,"is_preprint":false},{"pmid":"22354037","id":"PMC_22354037","title":"Genome-wide siRNA screen reveals amino acid starvation-induced autophagy requires SCOC and WAC.","date":"2012","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/22354037","citation_count":101,"is_preprint":false},{"pmid":"19289792","id":"PMC_19289792","title":"Wac: a new Augmin subunit required for chromosome alignment but not for acentrosomal microtubule assembly in female meiosis.","date":"2009","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/19289792","citation_count":59,"is_preprint":false},{"pmid":"12192034","id":"PMC_12192034","title":"Binding of Acf1 to DNA involves a WAC motif and is important for ACF-mediated chromatin assembly.","date":"2002","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/12192034","citation_count":52,"is_preprint":false},{"pmid":"26264232","id":"PMC_26264232","title":"WAC loss-of-function mutations cause a recognisable syndrome characterised by dysmorphic features, developmental delay and hypotonia and recapitulate 10p11.23 microdeletion syndrome.","date":"2015","source":"Journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/26264232","citation_count":48,"is_preprint":false},{"pmid":"28690313","id":"PMC_28690313","title":"The HDAC inhibitor panobinostat (LBH589) exerts in vivo anti-leukaemic activity against MLL-rearranged acute lymphoblastic leukaemia and involves the RNF20/RNF40/WAC-H2B ubiquitination axis.","date":"2017","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/28690313","citation_count":48,"is_preprint":false},{"pmid":"7932704","id":"PMC_7932704","title":"Fibritin encoded by bacteriophage T4 gene wac has a parallel triple-stranded alpha-helical coiled-coil structure.","date":"1994","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/7932704","citation_count":43,"is_preprint":false},{"pmid":"37532764","id":"PMC_37532764","title":"Small extracellular vesicles delivering lncRNA WAC-AS1 aggravate renal allograft ischemia‒reperfusion injury by inducing ferroptosis propagation.","date":"2023","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/37532764","citation_count":40,"is_preprint":false},{"pmid":"34527595","id":"PMC_34527595","title":"Identification of Glycolysis-Related lncRNAs and the Novel lncRNA WAC-AS1 Promotes Glycolysis and Tumor Progression in Hepatocellular Carcinoma.","date":"2021","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/34527595","citation_count":39,"is_preprint":false},{"pmid":"21811234","id":"PMC_21811234","title":"VCIP135 deubiquitinase and its binding protein, WAC, in p97ATPase-mediated membrane fusion.","date":"2011","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/21811234","citation_count":36,"is_preprint":false},{"pmid":"26757981","id":"PMC_26757981","title":"De novo loss-of-function mutations in WAC cause a recognizable intellectual disability syndrome and learning deficits in Drosophila.","date":"2016","source":"European journal of human genetics : EJHG","url":"https://pubmed.ncbi.nlm.nih.gov/26757981","citation_count":35,"is_preprint":false},{"pmid":"11827461","id":"PMC_11827461","title":"WAC, a novel WW domain-containing adapter with a coiled-coil region, is colocalized with splicing factor SC35.","date":"2002","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/11827461","citation_count":27,"is_preprint":false},{"pmid":"11895945","id":"PMC_11895945","title":"Transfer of the core region genes of the Yersinia enterocolitica WA-C serotype O:8 high-pathogenicity island to Y. enterocolitica MRS40, a strain with low levels of pathogenicity, confers a yersiniabactin biosynthesis phenotype and enhanced mouse virulence.","date":"2002","source":"Infection and immunity","url":"https://pubmed.ncbi.nlm.nih.gov/11895945","citation_count":22,"is_preprint":false},{"pmid":"33857290","id":"PMC_33857290","title":"Familial thrombocytopenia due to a complex structural variant resulting in a WAC-ANKRD26 fusion transcript.","date":"2021","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/33857290","citation_count":22,"is_preprint":false},{"pmid":"30021153","id":"PMC_30021153","title":"WAC Promotes Polo-like Kinase 1 Activation for Timely Mitotic 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SOX2.","date":"2023","source":"Biology direct","url":"https://pubmed.ncbi.nlm.nih.gov/37957698","citation_count":10,"is_preprint":false},{"pmid":"36053263","id":"PMC_36053263","title":"Long non-coding RNA WAC antisense RNA 1 mediates hepatitis B virus replication <em>in vitro</em> by reinforcing miR-192-5p/ATG7-induced autophagy.","date":"2022","source":"European journal of histochemistry : EJH","url":"https://pubmed.ncbi.nlm.nih.gov/36053263","citation_count":9,"is_preprint":false},{"pmid":"37106788","id":"PMC_37106788","title":"Structure-Function of the Human WAC Protein in GABAergic Neurons: Towards an Understanding of Autosomal Dominant DeSanto-Shinawi Syndrome.","date":"2023","source":"Biology","url":"https://pubmed.ncbi.nlm.nih.gov/37106788","citation_count":7,"is_preprint":false},{"pmid":"35018708","id":"PMC_35018708","title":"Clinical and molecular characterization of five new individuals with WAC-related intellectual disability: Evidence of pathogenicity for a novel splicing 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Part A","url":"https://pubmed.ncbi.nlm.nih.gov/34997803","citation_count":4,"is_preprint":false},{"pmid":"35766809","id":"PMC_35766809","title":"Phenotypic and Brain Imaging Findings Associated With a 10p Proximal Deletion Including the WAC Gene: Case Report and Literature Review.","date":"2022","source":"Cognitive and behavioral neurology : official journal of the Society for Behavioral and Cognitive Neurology","url":"https://pubmed.ncbi.nlm.nih.gov/35766809","citation_count":4,"is_preprint":false},{"pmid":"28820982","id":"PMC_28820982","title":"Direct analysis - no sample preparation - of bioavailable cortisol in human plasma by weak affinity chromatography (WAC).","date":"2017","source":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/28820982","citation_count":3,"is_preprint":false},{"pmid":"37702844","id":"PMC_37702844","title":"A novel anticancer quinolone, (R)-WAC-224, has anti-leukemia activities against acute myeloid leukemia.","date":"2023","source":"Investigational new drugs","url":"https://pubmed.ncbi.nlm.nih.gov/37702844","citation_count":2,"is_preprint":false},{"pmid":"37171241","id":"PMC_37171241","title":"The histone H2B ubiquitination regulator Wac is essential for plasma cell differentiation.","date":"2023","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/37171241","citation_count":2,"is_preprint":false},{"pmid":"37402055","id":"PMC_37402055","title":"Expression analyses of WAC, a responsible gene for neurodevelopmental disorders, during mouse brain development.","date":"2023","source":"Medical molecular morphology","url":"https://pubmed.ncbi.nlm.nih.gov/37402055","citation_count":1,"is_preprint":false},{"pmid":"40653822","id":"PMC_40653822","title":"Characterization of WAC interactions with R2TP and TTT chaperone complexes linking glucose and glutamine availability to mTORC1 activity.","date":"2025","source":"FEBS open bio","url":"https://pubmed.ncbi.nlm.nih.gov/40653822","citation_count":1,"is_preprint":false},{"pmid":"40341200","id":"PMC_40341200","title":"(R)-WAC-224, a new anticancer quinolone, combined with venetoclax and azacitidine overcomes venetoclax-resistant AML through MCL-1 downregulation.","date":"2025","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/40341200","citation_count":1,"is_preprint":false},{"pmid":"40893665","id":"PMC_40893665","title":"Inhibition of WAC alleviates the chondrocyte proinflammatory secretory phenotype and cartilage degradation via H2BK120ub1 and H3K27me3 coregulation.","date":"2025","source":"Acta pharmaceutica Sinica. 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WAC interacts with RNF20/40 via its C-terminal coiled-coil region and promotes RNF20/40's E3 ligase activity for histone H2B ubiquitination. The N-terminal WW domain of WAC recognizes RNA polymerase II, enabling WAC to target RNF20/40 to associate with the RNA polymerase II complex at active transcription sites.\",\n      \"method\": \"Protein affinity purification, Co-IP, domain mutagenesis, siRNA knockdown, H2B ubiquitination assay\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — multiple orthogonal methods (affinity purification, domain mutagenesis, functional ubiquitination assay) in a highly-cited foundational paper\",\n      \"pmids\": [\"21329877\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"WAC-dependent H2B ubiquitination and transcription regulation is important for cell-cycle checkpoint activation in response to genotoxic stress, as depletion of WAC abolished H2B ubiquitination and compromised checkpoint activation.\",\n      \"method\": \"siRNA knockdown, genotoxic stress assays, H2B ubiquitination assay\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean knockdown with defined cellular phenotype, supported by multiple assays in a highly-cited paper\",\n      \"pmids\": [\"21329877\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"WAC localizes to the Golgi and nucleus and was identified as a VCIP135-binding protein. WAC directly binds VCIP135 and increases its deubiquitinating activity, and WAC is required for p97/p47-mediated Golgi membrane fusion (but not p97/p37-mediated reassembly or ER membrane fusion).\",\n      \"method\": \"Co-IP, siRNA knockdown, in vitro Golgi reformation assay, deubiquitinase activity assay\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — direct binding shown, enzymatic activity assay performed, in vitro reconstitution of Golgi reformation, clean siRNA knockdown with specific pathway phenotype\",\n      \"pmids\": [\"21811234\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"WAC is required for starvation-induced autophagosome formation, identified in a genome-wide siRNA screen; WAC also acts as a potential negative regulator of the ubiquitin-proteasome system.\",\n      \"method\": \"Genome-wide siRNA screen, GFP-LC3 autophagosome marker assay, siRNA knockdown validation\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genome-wide screen with stringent validation criteria, multiple orthogonal validation steps, replicated in follow-up studies\",\n      \"pmids\": [\"22354037\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"WAC is bound to the Golgi by GM130 (a direct interaction required for autophagy). WAC and GM130 interact with the Atg8 homolog GABARAP and regulate its subcellular localization: GABARAP resides on the pericentriolar matrix, and WAC suppresses GM130 binding to GABARAP, enabling starvation-induced delivery of centrosomal GABARAP to the phagophore to activate ULK kinase.\",\n      \"method\": \"Co-IP, pulldown, siRNA knockdown, live-cell imaging, subcellular fractionation, ULK kinase activation assay\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — direct interaction established, localization studies with functional consequence, multiple orthogonal methods in a highly-cited paper\",\n      \"pmids\": [\"26687599\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"WAC was identified as a novel WW domain-containing adaptor protein that colocalizes with splicing factor SC35 (a marker for pre-mRNA splicing machinery) and exists mainly in a tyrosine-phosphorylated form in cells.\",\n      \"method\": \"Immunofluorescence, domain analysis, phosphorylation analysis\",\n      \"journal\": \"Genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — localization by immunofluorescence without deep functional follow-up; single method for colocalization\",\n      \"pmids\": [\"11827461\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"WAC promotes Polo-like kinase 1 (Plk1) activation for timely mitotic entry: Cdk1 phosphorylates WAC, priming its direct interaction with the polo-box domain of Plk1. WAC also binds Aurora A kinase (AurkA) and can enhance Plk1 phosphorylation by AurkA in vitro. Knockdown of WAC compromises Plk1 activity and delays mitotic entry, rescued by wild-type but not Plk1-binding-deficient WAC.\",\n      \"method\": \"Co-IP, in vitro kinase assay, mutagenesis, siRNA knockdown with rescue experiment, mitotic entry assay\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — in vitro kinase assay, direct interaction established, mutagenesis rescue experiment, clean loss-of-function phenotype\",\n      \"pmids\": [\"30021153\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"WAC functions as part of the RNF20/RNF40/WAC E3 ligase complex for H2B ubiquitination in leukemia cells; knockdown of WAC phenocopied loss of H2B ubiquitination and induced cell death in MLL-rearranged ALL.\",\n      \"method\": \"siRNA knockdown, H2B ubiquitination assay, cell viability assay\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean knockdown with specific molecular and cellular phenotype, consistent with foundational mechanistic work\",\n      \"pmids\": [\"28690313\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"B cell-specific Wac knockout mice show severely compromised T cell-dependent and -independent antibody responses, with drastically reduced plasma cell differentiation but intact germinal center B cell responses, linked to significant reduction in global H2B ubiquitination (ubH2B) in Wac-deficient B cells.\",\n      \"method\": \"Conditional B cell-specific knockout mouse model, antibody response assays, ubH2B western blotting, flow cytometry\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean conditional KO with defined cellular phenotype and molecular mechanism (ubH2B reduction)\",\n      \"pmids\": [\"37171241\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"WAC interacts with the transmembrane (TM) domains of PINK1 and prevents ubiquitination at the K137 site of PINK1, protecting PINK1 from ubiquitination-dependent degradation and thereby activating mitophagy to promote MSC osteogenesis.\",\n      \"method\": \"Co-IP, ubiquitination assay, mutagenesis (K137 site), in vitro and in vivo osteogenesis assays\",\n      \"journal\": \"Advanced science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct interaction and site-specific ubiquitination shown, functional rescue in osteogenesis; single lab\",\n      \"pmids\": [\"39555688\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"WAC directly binds mTOR-mLST8, R2TP, and TELO2 (but not TTI1 and TTI2) using purified proteins. In cells, WAC is part of complexes containing mTORC1, R2TP, and TTT components, and WAC and TELO2 strongly associate with mTOR under glucose and glutamine deprivation, with these interactions weakened upon nutrient refeeding, correlating with mTORC1 activity changes.\",\n      \"method\": \"Purified protein binding assays, Co-IP in cells under nutrient stress, transcriptomics, proteomics\",\n      \"journal\": \"FEBS open bio\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 — direct binding established with purified proteins, in-cell nutrient-responsive dynamics measured; single study\",\n      \"pmids\": [\"40653822\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Crystal structure of the yeast Bre1-Lge1 complex and AlphaFold-predicted structure of human RNF20/RNF40-WAC, combined with in vitro and in vivo experiments, revealed extensive interaction interfaces and identified key electrostatic interactions critical for binding specificity and H2B ubiquitination activity.\",\n      \"method\": \"X-ray crystallography (Bre1-Lge1), AlphaFold structure prediction (RNF20/RNF40-WAC), mutagenesis, in vitro ubiquitination assay, in vivo functional assays\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — crystal structure of orthologous complex, structure-guided mutagenesis validated in vitro and in vivo\",\n      \"pmids\": [\"41533567\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"WAC regulates H2BK120ub1 and influences H3K27me3 levels in chondrocytes by regulating nuclear entry of the H3K27 demethylase KDM6B, acting as a key crosstalk factor between H2BK120ub1 and H3K27me3 to control inflammatory mediator secretion and cartilage degradation.\",\n      \"method\": \"Cartilage-specific WAC knockout mice, H2BK120ub1 and H3K27me3 ChIP/western blotting, KDM6B nuclear entry assays, CIA and CIOA mouse models\",\n      \"journal\": \"Acta pharmaceutica sinica. B\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — conditional KO with defined molecular phenotype (crosstalk between histone marks), in vivo validation; single lab\",\n      \"pmids\": [\"40893665\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"WAC protein is expressed in a developmental stage-dependent manner in mouse brain and localizes predominantly to the perinuclear region of cortical neurons embryonically, then becomes enriched in the nucleus after birth; WAC is also present in axons and dendrites of cultured hippocampal neurons in a time-dependent manner.\",\n      \"method\": \"Anti-WAC antibody generation, western blotting, immunohistochemistry, immunofluorescence of primary cultured neurons\",\n      \"journal\": \"Medical molecular morphology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — direct localization with developmental context, but functional consequence not directly demonstrated in this study\",\n      \"pmids\": [\"37402055\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"A nuclear localization domain in WAC impacts the cellular distribution of the protein in cortical GABAergic neurons, as shown by deletion analysis of human protein domains.\",\n      \"method\": \"Domain deletion constructs, cellular distribution assay in GABAergic neurons\",\n      \"journal\": \"Biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — functional domain deletion with localization readout; single study\",\n      \"pmids\": [\"37106788\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"WAC is a WW domain- and coiled-coil-containing adaptor protein that promotes histone H2B ubiquitination by binding RNF20/40 (via its coiled-coil) and recruiting this E3 ligase complex to RNA polymerase II at active transcription sites (via its WW domain); it also regulates starvation-induced autophagy by modulating GABARAP localization between the Golgi (where GM130 sequesters it) and the centrosome/phagophore to activate ULK kinase, activates Plk1 for mitotic entry after Cdk1-mediated phosphorylation, protects PINK1 from ubiquitination-dependent degradation to facilitate mitophagy, and interacts with mTOR-mLST8, R2TP, and TELO2 to regulate mTORC1 activity in response to nutrient availability.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"WAC is a WW domain- and coiled-coil-containing adaptor protein that functions as a central scaffold linking chromatin modification, autophagy, cell-cycle progression, and nutrient signaling. Its best-characterized role is promoting histone H2B monoubiquitination by binding the RNF20/RNF40 E3 ligase complex through its coiled-coil domain and recruiting it to RNA polymerase II at active transcription sites via its WW domain, a function critical for transcription-coupled checkpoint activation, plasma cell differentiation, and histone mark crosstalk with H3K27me3 through regulation of KDM6B nuclear entry [PMID:21329877, PMID:37171241, PMID:40893665, PMID:41533567]. WAC also regulates starvation-induced autophagy by competing with GM130 for GABARAP binding at the Golgi, thereby enabling delivery of centrosomal GABARAP to the phagophore to activate ULK kinase, and separately promotes mitophagy by protecting PINK1 from ubiquitination-dependent degradation [PMID:26687599, PMID:39555688]. Additionally, WAC facilitates timely mitotic entry by serving as a Cdk1-phosphorylated scaffold that bridges Aurora A and Plk1, participates in p97/p47-mediated Golgi membrane fusion by binding and activating the deubiquitinase VCIP135, and associates with mTOR-mLST8/R2TP/TELO2 complexes in a nutrient-responsive manner to modulate mTORC1 activity [PMID:30021153, PMID:21811234, PMID:40653822].\",\n  \"teleology\": [\n    {\n      \"year\": 2002,\n      \"claim\": \"Initial identification of WAC as a novel WW domain-containing adaptor established its domain architecture and suggested a connection to nuclear RNA processing through colocalization with splicing factor SC35.\",\n      \"evidence\": \"Immunofluorescence and domain analysis in cultured cells\",\n      \"pmids\": [\"11827461\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Colocalization with SC35 shown by single method without functional consequence demonstrated\", \"No interacting partners identified\", \"Tyrosine phosphorylation significance unknown\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"The discovery that WAC bridges RNF20/RNF40 to RNA polymerase II via distinct domains to promote H2B ubiquitination established WAC's primary molecular function as a transcription-coupled chromatin modification scaffold, and linked this activity to genotoxic stress checkpoint activation.\",\n      \"evidence\": \"Affinity purification, Co-IP, domain mutagenesis, siRNA knockdown, H2B ubiquitination and checkpoint assays in human cells\",\n      \"pmids\": [\"21329877\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of WAC-RNF20/40 interaction not resolved at atomic level\", \"In vivo physiological relevance in organism not tested\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Parallel work revealed a second function for WAC at the Golgi: direct binding and activation of the deubiquitinase VCIP135 for p97/p47-dependent Golgi membrane fusion, establishing WAC as a bifunctional nuclear-Golgi protein.\",\n      \"evidence\": \"Co-IP, in vitro Golgi reformation assay, DUB activity assay, siRNA knockdown\",\n      \"pmids\": [\"21811234\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How WAC partitions between nuclear and Golgi functions is unclear\", \"Ubiquitin substrates of VCIP135 at the Golgi not identified\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"A genome-wide siRNA screen identified WAC as required for starvation-induced autophagosome formation, opening a third functional axis beyond chromatin and Golgi membrane dynamics.\",\n      \"evidence\": \"Genome-wide siRNA screen with GFP-LC3 reporter validation\",\n      \"pmids\": [\"22354037\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism connecting WAC to autophagy not yet defined\", \"Potential indirect effects through H2B ubiquitination not excluded\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"The autophagy mechanism was resolved: WAC is tethered to the Golgi by GM130 and competes with GM130 for GABARAP binding, enabling starvation-triggered release of centrosomal GABARAP to phagophores where it activates ULK kinase.\",\n      \"evidence\": \"Co-IP, pulldown, live-cell imaging, subcellular fractionation, ULK kinase activation assay\",\n      \"pmids\": [\"26687599\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Signal that switches WAC from GM130-bound to GABARAP-releasing state not identified\", \"Whether WAC regulation of GABARAP is tissue-specific is unknown\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"WAC was shown to promote mitotic entry by acting as a Cdk1-phosphorylated scaffold that recruits Plk1 via its polo-box domain and enhances Plk1 phosphorylation by Aurora A, revealing a cell-cycle regulatory function.\",\n      \"evidence\": \"In vitro kinase assays, Co-IP, mutagenesis with rescue experiments, mitotic entry assays\",\n      \"pmids\": [\"30021153\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether WAC's mitotic role is coordinated with its chromatin or autophagy functions is unknown\", \"Structural details of the WAC-Plk1-AurkA ternary complex not resolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Conditional B cell-specific Wac knockout demonstrated that WAC-dependent H2B ubiquitination is essential in vivo for plasma cell differentiation and antibody responses, while germinal center formation is spared.\",\n      \"evidence\": \"Conditional knockout mouse model, flow cytometry, antibody response assays, ubH2B western blotting\",\n      \"pmids\": [\"37171241\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Transcriptional targets downstream of WAC-dependent ubH2B in plasma cells not identified\", \"Whether other H2B ubiquitination-independent WAC functions contribute to the B cell phenotype is untested\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Characterization of WAC protein expression in developing mouse brain showed stage-dependent localization from perinuclear (embryonic) to nuclear (postnatal) in cortical neurons, with a nuclear localization domain identified by deletion analysis.\",\n      \"evidence\": \"Immunohistochemistry, immunofluorescence, domain deletion constructs in neurons\",\n      \"pmids\": [\"37402055\", \"37106788\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of developmental relocalization not demonstrated\", \"Domain deletion studies performed only in GABAergic neurons\", \"No loss-of-function neuronal phenotype directly shown\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"WAC was found to protect PINK1 from ubiquitination-dependent degradation by binding its transmembrane domain and blocking K137 ubiquitination, thereby activating mitophagy to promote MSC osteogenesis.\",\n      \"evidence\": \"Co-IP, ubiquitination assay with K137 mutagenesis, in vitro and in vivo osteogenesis assays\",\n      \"pmids\": [\"39555688\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab finding not independently replicated\", \"Whether WAC-PINK1 interaction occurs broadly or is specific to MSC context is unclear\", \"E3 ligase targeting K137 not identified\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Direct binding assays with purified proteins established that WAC interacts with mTOR-mLST8, R2TP, and TELO2 in a nutrient-responsive manner, with associations strengthened during glucose/glutamine deprivation and weakened upon refeeding, linking WAC to mTORC1 regulation.\",\n      \"evidence\": \"Purified protein binding assays, Co-IP under nutrient stress, proteomics\",\n      \"pmids\": [\"40653822\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single study; independent replication needed\", \"Functional consequence of WAC on mTORC1 signaling output not directly demonstrated\", \"Relationship to WAC's autophagy function via GABARAP/ULK pathway not resolved\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Cartilage-specific WAC knockout revealed that WAC-dependent H2BK120ub1 regulates H3K27me3 levels by controlling nuclear entry of the demethylase KDM6B, establishing WAC as a histone mark crosstalk regulator with consequences for inflammatory cartilage degradation.\",\n      \"evidence\": \"Conditional knockout mice, ChIP, western blotting, KDM6B nuclear entry assays, arthritis mouse models\",\n      \"pmids\": [\"40893665\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which H2BK120ub1 controls KDM6B nuclear entry not molecularly defined\", \"Single-lab finding\", \"Whether this crosstalk operates in other tissues unknown\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Crystal structure of the yeast Bre1-Lge1 complex and AlphaFold-predicted structure of human RNF20/RNF40-WAC resolved the interaction interfaces and identified electrostatic contacts critical for binding specificity and H2B ubiquitination, providing the first structural framework for this complex.\",\n      \"evidence\": \"X-ray crystallography of yeast orthologs, AlphaFold prediction for human complex, structure-guided mutagenesis with in vitro and in vivo validation\",\n      \"pmids\": [\"41533567\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Human complex structure predicted rather than experimentally determined\", \"No structure of WAC WW domain bound to RNA Pol II CTD\", \"Structural basis of WAC's non-chromatin functions unexplored\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How WAC coordinates its multiple scaffold functions—chromatin modification, autophagy, mitotic entry, Golgi membrane fusion, mTORC1 signaling, and mitophagy—in a cell-state-dependent manner remains an open question, as does the upstream regulatory logic that partitions WAC among these pathways.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No integrative study examining how WAC's multiple functions are coordinated or mutually exclusive\", \"Full post-translational modification map of WAC not established\", \"No disease-causative mutations in WAC identified through human genetic studies in the timeline\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 4, 6, 10]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [2, 9]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 5, 13, 14]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [2, 4]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [6, 10]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [0, 1, 7, 8, 11, 12]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [3, 4]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [6]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [0, 1, 12]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [10]}\n    ],\n    \"complexes\": [\n      \"RNF20/RNF40/WAC E3 ligase complex\",\n      \"mTORC1-R2TP-TTT complex\"\n    ],\n    \"partners\": [\n      \"RNF20\",\n      \"RNF40\",\n      \"GM130\",\n      \"GABARAP\",\n      \"PLK1\",\n      \"VCIP135\",\n      \"PINK1\",\n      \"TELO2\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}