{"gene":"WAC","run_date":"2026-06-11T09:02:06","timeline":{"discoveries":[{"year":2011,"finding":"WAC was identified as a functional partner of RNF20/40 E3 ligase complex. WAC interacts with RNF20/40 through 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, and WAC targets RNF20/40 to associate with the RNA polymerase II complex at active transcription sites. Depletion of WAC abolishes H2B ubiquitination.","method":"Protein affinity purification, Co-IP, domain mutational analysis, siRNA depletion with H2B ubiquitination readout","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — affinity purification, domain mapping, functional rescue, replicated in multiple downstream studies","pmids":["21329877"],"is_preprint":false},{"year":2015,"finding":"WAC and GM130 are both required for starvation-induced autophagy. GM130 directly interacts with WAC and tethers it to the Golgi. WAC and GM130 interact with the Atg8 homolog GABARAP and regulate its subcellular localization; GABARAP resides on the pericentriolar matrix and this pool contributes to autophagosome formation. WAC suppresses GM130 binding to GABARAP, enabling starvation-induced centrosomal GABARAP delivery to the phagophore to activate ULK kinase via the ULK1 LIR motif.","method":"Co-IP, siRNA knockdown, live-cell imaging, subcellular fractionation, in vitro ULK kinase assay, LIR motif mutagenesis","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal methods including in vitro kinase assay, mutagenesis, imaging, and Co-IP in a single rigorous study","pmids":["26687599"],"is_preprint":false},{"year":2012,"finding":"WAC is required for amino acid starvation-induced autophagy (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 using GFP-LC3 reporter cell line, validation by siRNA knockdown","journal":"The EMBO journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genome-wide screen with stringent validation criteria, single lab, autophagy phenotype confirmed but molecular mechanism not fully resolved","pmids":["22354037"],"is_preprint":false},{"year":2011,"finding":"WAC localizes to both the Golgi and nucleus. At the Golgi, WAC is part of a complex containing VCIP135 (a deubiquitinase) and p97 ATPase. WAC directly binds VCIP135 and increases its deubiquitinating activity. WAC is required for p97/p47-mediated Golgi membrane fusion (Golgi biogenesis) but not for p97/p37-mediated reassembly, and is dispensable for p97-mediated ER membrane fusion.","method":"Co-IP, siRNA knockdown, in vitro Golgi reformation assay, in vitro deubiquitinase activity assay, immunofluorescence","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro biochemical activity assay combined with siRNA knockdown and reconstitution assay, multiple orthogonal methods","pmids":["21811234"],"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 enhances Plk1 phosphorylation by AurkA in vitro. Knockdown of WAC compromises Plk1 activity and delays mitotic entry; these defects are rescued by wild-type WAC but not by a Plk1-binding-deficient mutant.","method":"Co-IP, in vitro kinase assay, domain mutational analysis (Plk1-binding-deficient mutant), siRNA knockdown with mitotic entry/Plk1 activity readout, rescue experiment","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro kinase assay, structure-function mutagenesis, rescue experiment with binding-deficient mutant, single lab but multiple orthogonal methods","pmids":["30021153"],"is_preprint":false},{"year":2002,"finding":"WAC (identified initially as a WW domain-containing adaptor) colocalizes with splicing factor SC35 by immunofluorescence, suggesting a role in pre-mRNA splicing. WAC existed mainly in a tyrosine-phosphorylated form.","method":"Immunofluorescence colocalization, domain analysis (Rosetta stone/domain fusion bioinformatics), phosphorylation detection","journal":"Genomics","confidence":"Low","confidence_rationale":"Tier 3 / Weak — colocalization by immunofluorescence only, no functional consequence demonstrated, single lab, single method","pmids":["11827461"],"is_preprint":false},{"year":2017,"finding":"Knockdown of WAC phenocopies loss of H2B ubiquitination in MLL-rearranged ALL cells and causes cell death, demonstrating that the RNF20/RNF40/WAC E3 ligase complex is a pivotal pathway for MLL-rearranged leukaemic cell maintenance. HDAC inhibitor panobinostat suppresses this complex activity.","method":"siRNA knockdown of WAC with H2B ubiquitination and cell viability readout, pharmacological inhibition","journal":"Leukemia","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — WAC knockdown with defined molecular and cellular phenotype corroborating the RNF20/40/WAC pathway, single lab","pmids":["28690313"],"is_preprint":false},{"year":2024,"finding":"WAC protects PINK1 (a key initiator of mitophagy) from ubiquitination-dependent degradation by interacting with the transmembrane (TM) domains of PINK1 and preventing ubiquitination at PINK1's K137 site. This stabilization of PINK1 activates mitophagy and promotes MSC osteogenic differentiation.","method":"Co-IP, ubiquitination assay, site-directed mutagenesis (K137 site), knockdown/overexpression with mitophagy and osteogenesis readout, in vitro and in vivo bone formation assays","journal":"Advanced science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, site-specific ubiquitination mutagenesis, in vitro and in vivo functional assays, single lab","pmids":["39555688"],"is_preprint":false},{"year":2023,"finding":"WAC is essential for plasma cell (PC) differentiation in B cells. B cell-specific Wac knockout mice show severely compromised antibody responses and drastically reduced PC differentiation. Wac deficiency leads to a significant reduction in global histone H2B ubiquitination (ubH2B) in B cells, correlated with downregulated expression of genes critical for cell metabolism.","method":"B cell-specific conditional knockout mouse, flow cytometry for PC differentiation, H2B ubiquitination western blot, antibody response assay","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean conditional KO with defined cellular and molecular phenotype (H2BUb1 reduction), single lab","pmids":["37171241"],"is_preprint":false},{"year":2025,"finding":"WAC directly binds mTOR-mLST8, R2TP chaperone complex, and TELO2 (component of TTT chaperone), but not TTI1 or TTI2, as established with purified proteins. In cells, WAC forms complexes containing mTORC1, R2TP, and TTT components that are modulated by nutrient availability. WAC and TELO2 strongly associate with mTOR under glucose and glutamine deprivation, and these interactions are weakened after nutrient refeeding, correlating with changes in mTORC1 activity.","method":"In vitro binding assay with purified proteins, Co-IP in cells under nutrient-defined conditions, transcriptomic and proteomic analysis","journal":"FEBS open bio","confidence":"Medium","confidence_rationale":"Tier 1–2 / Moderate — direct binding established with purified proteins and Co-IP in cells under defined nutrient conditions, single lab, multiple methods","pmids":["40653822"],"is_preprint":false},{"year":2026,"finding":"Crystal structure of the yeast Bre1-Lge1 complex and AlphaFold-predicted structure of the human RNF20/RNF40-WAC complex were determined and validated by in vitro and in vivo experiments. Extensive RNF20/RNF40-WAC interfaces were identified with key electrostatic interactions encoding binding specificity. These interactions are critical for H2BUb1 catalysis and the processes it regulates. The RNF20/RNF40-WAC and Bre1-Lge1 interfaces share structural homology but use different electrostatic interactions.","method":"X-ray crystallography (Bre1-Lge1), AlphaFold structural modeling (RNF20/RNF40-WAC), in vitro and in vivo mutagenesis and functional assays","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure plus AlphaFold with experimental validation by mutagenesis and functional assays, single lab but multiple orthogonal methods","pmids":["41533567"],"is_preprint":false},{"year":2025,"finding":"WAC regulates cartilage degradation in arthritis by controlling H2BK120ub1 levels and influencing H3K27me3 through regulation of nuclear entry of the H3K27 demethylase KDM6B. WAC acts as a key factor in crosstalk between H2BK120ub1 and H3K27me3. Cartilage-specific knockout of WAC alleviates cartilage degradation in collagen-induced arthritis and collagenase-induced osteoarthritis mouse models.","method":"Cartilage-specific conditional knockout mouse, ChIP for H2BK120ub1 and H3K27me3, nuclear fractionation for KDM6B localization, in vitro and in vivo arthritis models","journal":"Acta pharmaceutica sinica. B","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — conditional KO with defined epigenetic and cellular phenotypes, mechanistic link to KDM6B nuclear entry, single lab","pmids":["40893665"],"is_preprint":false},{"year":2016,"finding":"Loss-of-function of the Drosophila WAC orthologue (CG8949) by neuronal knockdown impairs habituation learning in Drosophila, establishing that WAC is required in neurons for normal cognitive performance.","method":"Neuronal RNAi knockdown in Drosophila, habituation learning behavioral assay","journal":"European journal of human genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean neuronal-specific knockdown with defined behavioral phenotype in a well-established learning paradigm, single lab","pmids":["26757981"],"is_preprint":false},{"year":2023,"finding":"WAC protein localizes predominantly in the nucleus of cortical neurons after birth and in hippocampal neurons, but is found in the perinuclear region at earlier developmental stages. WAC is also detected in axons and dendrites in a time-dependent manner in primary cultured hippocampal neurons. Nuclear localization is enriched postnatally in cerebral cortex, hippocampus, and cerebellum.","method":"Immunohistochemistry, Western blotting with stage-specific brain samples, immunofluorescence in primary cultured neurons","journal":"Medical molecular morphology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — direct localization experiment across developmental stages, multiple brain regions, independent antibody preparation; no functional consequence directly linked","pmids":["37402055"],"is_preprint":false},{"year":2023,"finding":"A nuclear localization domain within WAC was identified and experimentally tested; deletion of this domain impacts the cellular distribution of the WAC protein in GABAergic neurons.","method":"Domain deletion constructs with fluorescence-based localization assay in cortical GABAergic neurons","journal":"Biology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, single method (fluorescence localization), no functional consequence beyond distribution change","pmids":["37106788"],"is_preprint":false}],"current_model":"WAC is a WW domain- and coiled-coil-containing adaptor protein that (1) promotes transcription-coupled histone H2B ubiquitination by binding RNA polymerase II via its WW domain and recruiting the RNF20/RNF40 E3 ligase complex via its coiled-coil region; (2) regulates starvation-induced autophagy by suppressing GM130-mediated Golgi tethering of GABARAP, enabling centrosomal GABARAP delivery to the phagophore to activate ULK kinase; (3) activates Polo-like kinase 1 for timely mitotic entry by bridging Cdk1-phosphorylated WAC with Plk1's polo-box domain and Aurora A; (4) enhances VCIP135 deubiquitinase activity within a p97/p47 Golgi membrane fusion pathway; (5) protects PINK1 from ubiquitin-dependent degradation to promote mitophagy; and (6) modulates mTORC1 activity through nutrient-sensitive interactions with mTOR-mLST8, R2TP, and TELO2."},"narrative":{"mechanistic_narrative":"WAC is a WW domain- and coiled-coil-containing adaptor protein that couples active transcription to chromatin modification and serves as a multifunctional scaffold across nuclear, Golgi, and autophagy compartments. Its best-characterized role is to drive transcription-coupled histone H2B monoubiquitination: its N-terminal WW domain engages RNA polymerase II while its C-terminal coiled-coil region binds and stimulates the RNF20/RNF40 E3 ligase, targeting the ligase to active transcription sites, and WAC loss abolishes H2B ubiquitination [PMID:21329877]. Structural modeling of the human RNF20/RNF40-WAC interface, validated by mutagenesis, defines electrostatic contacts that encode binding specificity essential for H2BUb1 catalysis [PMID:41533567]. This activity has physiological consequences: WAC is required for plasma cell differentiation and antibody responses through global H2B ubiquitination [PMID:37171241], sustains MLL-rearranged leukaemic cells [PMID:28690313], and controls cartilage degradation by linking H2BK120ub1 to H3K27me3 via regulation of KDM6B nuclear entry [PMID:40893665]. Independently of transcription, WAC regulates starvation-induced autophagy by suppressing GM130-mediated Golgi tethering of GABARAP, releasing a pericentriolar GABARAP pool to the phagophore to activate ULK kinase [PMID:26687599, PMID:22354037], and at the Golgi it binds the deubiquitinase VCIP135 and enhances its activity within the p97/p47 membrane fusion pathway [PMID:21811234]. WAC additionally promotes timely mitotic entry by bridging Cdk1-phosphorylated WAC to the Plk1 polo-box domain and Aurora A to activate Plk1 [PMID:30021153], protects PINK1 from ubiquitin-dependent degradation to promote mitophagy [PMID:39555688], and modulates mTORC1 through nutrient-sensitive interactions with mTOR-mLST8, R2TP, and TELO2 [PMID:40653822]. Neuronal WAC is required for normal habituation learning [PMID:26757981].","teleology":[{"year":2002,"claim":"An initial question was what cellular compartment and process this WW-domain adaptor belongs to; nuclear speckle colocalization first hinted at an RNA-processing connection.","evidence":"Immunofluorescence colocalization with splicing factor SC35 and phosphorylation detection","pmids":["11827461"],"confidence":"Low","gaps":["colocalization only, no functional consequence demonstrated","splicing role never mechanistically established in later work","tyrosine-phosphorylation significance unresolved"]},{"year":2011,"claim":"The central mechanistic role was established by showing WAC bridges RNA polymerase II to the RNF20/RNF40 ligase, defining it as the adaptor that couples transcription to H2B ubiquitination.","evidence":"Affinity purification, Co-IP, domain mutational analysis and siRNA depletion with H2B ubiquitination readout","pmids":["21329877"],"confidence":"High","gaps":["structural basis of the RNF20/40 interface not resolved at this stage","downstream transcriptional targets undefined"]},{"year":2011,"claim":"A distinct cytoplasmic function was revealed at the Golgi, where WAC was shown to enhance VCIP135 deubiquitinase activity and is selectively required for p97/p47-mediated Golgi membrane fusion.","evidence":"Co-IP, in vitro Golgi reformation and deubiquitinase activity assays, immunofluorescence","pmids":["21811234"],"confidence":"High","gaps":["how nuclear and Golgi pools are partitioned unknown","mechanism by which WAC stimulates VCIP135 unresolved"]},{"year":2012,"claim":"A genome-wide screen established WAC as a positive regulator of starvation-induced autophagosome formation, broadening its role beyond transcription.","evidence":"Genome-wide siRNA screen with GFP-LC3 reporter and knockdown validation","pmids":["22354037"],"confidence":"Medium","gaps":["molecular mechanism not resolved in the screen","link between autophagy role and UPS regulation undefined"]},{"year":2015,"claim":"The autophagy mechanism was resolved: WAC suppresses GM130 tethering of GABARAP to free a pericentriolar GABARAP pool for phagophore delivery and ULK activation.","evidence":"Co-IP, knockdown, live-cell imaging, fractionation, in vitro ULK kinase assay and LIR mutagenesis","pmids":["26687599"],"confidence":"High","gaps":["signal that triggers WAC-GM130 competition upon starvation unclear","relationship to WAC's nuclear function not addressed"]},{"year":2016,"claim":"Neuronal requirement was demonstrated by showing the Drosophila orthologue is needed for habituation learning, linking WAC to cognitive function.","evidence":"Neuronal RNAi knockdown in Drosophila with habituation behavioral assay","pmids":["26757981"],"confidence":"Medium","gaps":["molecular pathway in neurons not defined","which WAC activity underlies the learning phenotype unknown"]},{"year":2017,"claim":"The transcriptional ligase pathway was given disease relevance by showing WAC is required to maintain MLL-rearranged leukaemic cells via H2B ubiquitination.","evidence":"siRNA knockdown with H2B ubiquitination and viability readout plus pharmacological inhibition","pmids":["28690313"],"confidence":"Medium","gaps":["specific target genes driving leukaemic dependency undefined","selectivity over normal cells unaddressed"]},{"year":2018,"claim":"A cell-cycle function was established: Cdk1-phosphorylated WAC binds the Plk1 polo-box domain and Aurora A to activate Plk1 for timely mitotic entry.","evidence":"Co-IP, in vitro kinase assay, Plk1-binding-deficient mutant and rescue with mitotic entry readout","pmids":["30021153"],"confidence":"High","gaps":["how this mitotic role integrates with WAC's chromatin function unclear","structural basis of WAC-PBD interaction not solved"]},{"year":2023,"claim":"In vivo significance of the H2B ubiquitination role was shown via a B cell-specific knockout that abolishes plasma cell differentiation and reduces global ubH2B.","evidence":"B cell-specific conditional knockout mouse, flow cytometry, ubH2B western blot, antibody response assay","pmids":["37171241"],"confidence":"Medium","gaps":["specific metabolic target genes regulated not fully mapped","whether non-transcriptional WAC roles contribute unaddressed"]},{"year":2023,"claim":"Neuronal localization was characterized as developmentally dynamic, predominantly nuclear postnatally and perinuclear/axodendritic earlier, with a defined nuclear localization domain.","evidence":"Immunohistochemistry, Western blotting across brain stages, and domain-deletion fluorescence assays in neurons","pmids":["37402055","37106788"],"confidence":"Medium","gaps":["functional consequence of subcellular redistribution not established","regulation of the nuclear localization domain unknown"]},{"year":2024,"claim":"A mitophagy function was identified: WAC binds PINK1 transmembrane domains and blocks K137 ubiquitination, stabilizing PINK1 to promote mitophagy and osteogenic differentiation.","evidence":"Co-IP, ubiquitination assay, K137 site mutagenesis, in vitro and in vivo bone formation assays","pmids":["39555688"],"confidence":"Medium","gaps":["E3 ligase WAC competes with not identified","relationship to GABARAP-dependent autophagy role unclear"]},{"year":2025,"claim":"A nutrient-sensing function was proposed by showing direct WAC binding to mTOR-mLST8, R2TP, and TELO2 in nutrient-modulated complexes correlating with mTORC1 activity.","evidence":"In vitro binding with purified proteins and Co-IP under defined nutrient conditions plus omics","pmids":["40653822"],"confidence":"Medium","gaps":["causal effect of WAC on mTORC1 output not demonstrated by loss-of-function","structural basis of binding unresolved"]},{"year":2025,"claim":"The epigenetic crosstalk role was extended by showing WAC controls H2BK120ub1 and influences H3K27me3 via KDM6B nuclear entry, with cartilage-specific knockout protecting against arthritis.","evidence":"Cartilage-specific conditional KO, ChIP for histone marks, nuclear fractionation for KDM6B, in vivo arthritis models","pmids":["40893665"],"confidence":"Medium","gaps":["mechanism by which WAC regulates KDM6B import undefined","direct gene targets of the ub-methyl crosstalk not enumerated"]},{"year":2026,"claim":"The structural basis of the core adaptor function was defined, mapping the RNF20/RNF40-WAC interface and its electrostatic specificity determinants required for H2BUb1 catalysis.","evidence":"X-ray crystallography of yeast Bre1-Lge1, AlphaFold modeling of human RNF20/40-WAC, and mutagenesis with functional validation","pmids":["41533567"],"confidence":"High","gaps":["full-length human complex not crystallized","how WW-domain Pol II engagement couples to ligase geometry unresolved"]},{"year":null,"claim":"It remains unknown how WAC's distinct nuclear (H2B ubiquitination), Golgi/autophagy, mitotic, and nutrient-sensing activities are coordinated within a single cell and which are regulated by the same post-translational or localization switches.","evidence":"","pmids":[],"confidence":"Medium","gaps":["no unifying regulatory model integrating compartment-specific functions","signals partitioning WAC between nucleus, Golgi, and centrosome undefined","in vivo loss-of-function for mTORC1 and mitophagy roles lacking"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,1,3,4]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,3,4,7]},{"term_id":"GO:0042393","term_label":"histone binding","supporting_discovery_ids":[0,10]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,3,13]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[1,3]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[1]}],"pathway":[{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[0,10,11]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[0,8]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[1,2,7]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[4]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[3]}],"complexes":["RNF20/RNF40-WAC H2B ubiquitination complex","p97/p47-VCIP135 Golgi membrane fusion complex"],"partners":["RNF20","RNF40","GM130","GABARAP","VCIP135","PLK1","PINK1","MTOR"],"other_free_text":[]}},"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":136,"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":118,"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":"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":51,"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":"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":42,"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 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WAC interacts with RNF20/40 through 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, and WAC targets RNF20/40 to associate with the RNA polymerase II complex at active transcription sites. Depletion of WAC abolishes H2B ubiquitination.\",\n      \"method\": \"Protein affinity purification, Co-IP, domain mutational analysis, siRNA depletion with H2B ubiquitination readout\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — affinity purification, domain mapping, functional rescue, replicated in multiple downstream studies\",\n      \"pmids\": [\"21329877\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"WAC and GM130 are both required for starvation-induced autophagy. GM130 directly interacts with WAC and tethers it to the Golgi. WAC and GM130 interact with the Atg8 homolog GABARAP and regulate its subcellular localization; GABARAP resides on the pericentriolar matrix and this pool contributes to autophagosome formation. WAC suppresses GM130 binding to GABARAP, enabling starvation-induced centrosomal GABARAP delivery to the phagophore to activate ULK kinase via the ULK1 LIR motif.\",\n      \"method\": \"Co-IP, siRNA knockdown, live-cell imaging, subcellular fractionation, in vitro ULK kinase assay, LIR motif mutagenesis\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal methods including in vitro kinase assay, mutagenesis, imaging, and Co-IP in a single rigorous study\",\n      \"pmids\": [\"26687599\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"WAC is required for amino acid starvation-induced autophagy (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 using GFP-LC3 reporter cell line, validation by siRNA knockdown\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genome-wide screen with stringent validation criteria, single lab, autophagy phenotype confirmed but molecular mechanism not fully resolved\",\n      \"pmids\": [\"22354037\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"WAC localizes to both the Golgi and nucleus. At the Golgi, WAC is part of a complex containing VCIP135 (a deubiquitinase) and p97 ATPase. WAC directly binds VCIP135 and increases its deubiquitinating activity. WAC is required for p97/p47-mediated Golgi membrane fusion (Golgi biogenesis) but not for p97/p37-mediated reassembly, and is dispensable for p97-mediated ER membrane fusion.\",\n      \"method\": \"Co-IP, siRNA knockdown, in vitro Golgi reformation assay, in vitro deubiquitinase activity assay, immunofluorescence\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro biochemical activity assay combined with siRNA knockdown and reconstitution assay, multiple orthogonal methods\",\n      \"pmids\": [\"21811234\"],\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 enhances Plk1 phosphorylation by AurkA in vitro. Knockdown of WAC compromises Plk1 activity and delays mitotic entry; these defects are rescued by wild-type WAC but not by a Plk1-binding-deficient mutant.\",\n      \"method\": \"Co-IP, in vitro kinase assay, domain mutational analysis (Plk1-binding-deficient mutant), siRNA knockdown with mitotic entry/Plk1 activity readout, rescue experiment\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro kinase assay, structure-function mutagenesis, rescue experiment with binding-deficient mutant, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"30021153\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"WAC (identified initially as a WW domain-containing adaptor) colocalizes with splicing factor SC35 by immunofluorescence, suggesting a role in pre-mRNA splicing. WAC existed mainly in a tyrosine-phosphorylated form.\",\n      \"method\": \"Immunofluorescence colocalization, domain analysis (Rosetta stone/domain fusion bioinformatics), phosphorylation detection\",\n      \"journal\": \"Genomics\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — colocalization by immunofluorescence only, no functional consequence demonstrated, single lab, single method\",\n      \"pmids\": [\"11827461\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Knockdown of WAC phenocopies loss of H2B ubiquitination in MLL-rearranged ALL cells and causes cell death, demonstrating that the RNF20/RNF40/WAC E3 ligase complex is a pivotal pathway for MLL-rearranged leukaemic cell maintenance. HDAC inhibitor panobinostat suppresses this complex activity.\",\n      \"method\": \"siRNA knockdown of WAC with H2B ubiquitination and cell viability readout, pharmacological inhibition\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — WAC knockdown with defined molecular and cellular phenotype corroborating the RNF20/40/WAC pathway, single lab\",\n      \"pmids\": [\"28690313\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"WAC protects PINK1 (a key initiator of mitophagy) from ubiquitination-dependent degradation by interacting with the transmembrane (TM) domains of PINK1 and preventing ubiquitination at PINK1's K137 site. This stabilization of PINK1 activates mitophagy and promotes MSC osteogenic differentiation.\",\n      \"method\": \"Co-IP, ubiquitination assay, site-directed mutagenesis (K137 site), knockdown/overexpression with mitophagy and osteogenesis readout, in vitro and in vivo bone formation assays\",\n      \"journal\": \"Advanced science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, site-specific ubiquitination mutagenesis, in vitro and in vivo functional assays, single lab\",\n      \"pmids\": [\"39555688\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"WAC is essential for plasma cell (PC) differentiation in B cells. B cell-specific Wac knockout mice show severely compromised antibody responses and drastically reduced PC differentiation. Wac deficiency leads to a significant reduction in global histone H2B ubiquitination (ubH2B) in B cells, correlated with downregulated expression of genes critical for cell metabolism.\",\n      \"method\": \"B cell-specific conditional knockout mouse, flow cytometry for PC differentiation, H2B ubiquitination western blot, antibody response assay\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean conditional KO with defined cellular and molecular phenotype (H2BUb1 reduction), single lab\",\n      \"pmids\": [\"37171241\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"WAC directly binds mTOR-mLST8, R2TP chaperone complex, and TELO2 (component of TTT chaperone), but not TTI1 or TTI2, as established with purified proteins. In cells, WAC forms complexes containing mTORC1, R2TP, and TTT components that are modulated by nutrient availability. WAC and TELO2 strongly associate with mTOR under glucose and glutamine deprivation, and these interactions are weakened after nutrient refeeding, correlating with changes in mTORC1 activity.\",\n      \"method\": \"In vitro binding assay with purified proteins, Co-IP in cells under nutrient-defined conditions, transcriptomic and proteomic analysis\",\n      \"journal\": \"FEBS open bio\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — direct binding established with purified proteins and Co-IP in cells under defined nutrient conditions, single lab, multiple methods\",\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 the human RNF20/RNF40-WAC complex were determined and validated by in vitro and in vivo experiments. Extensive RNF20/RNF40-WAC interfaces were identified with key electrostatic interactions encoding binding specificity. These interactions are critical for H2BUb1 catalysis and the processes it regulates. The RNF20/RNF40-WAC and Bre1-Lge1 interfaces share structural homology but use different electrostatic interactions.\",\n      \"method\": \"X-ray crystallography (Bre1-Lge1), AlphaFold structural modeling (RNF20/RNF40-WAC), in vitro and in vivo mutagenesis and functional assays\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure plus AlphaFold with experimental validation by mutagenesis and functional assays, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"41533567\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"WAC regulates cartilage degradation in arthritis by controlling H2BK120ub1 levels and influencing H3K27me3 through regulation of nuclear entry of the H3K27 demethylase KDM6B. WAC acts as a key factor in crosstalk between H2BK120ub1 and H3K27me3. Cartilage-specific knockout of WAC alleviates cartilage degradation in collagen-induced arthritis and collagenase-induced osteoarthritis mouse models.\",\n      \"method\": \"Cartilage-specific conditional knockout mouse, ChIP for H2BK120ub1 and H3K27me3, nuclear fractionation for KDM6B localization, in vitro and in vivo arthritis models\",\n      \"journal\": \"Acta pharmaceutica sinica. B\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — conditional KO with defined epigenetic and cellular phenotypes, mechanistic link to KDM6B nuclear entry, single lab\",\n      \"pmids\": [\"40893665\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Loss-of-function of the Drosophila WAC orthologue (CG8949) by neuronal knockdown impairs habituation learning in Drosophila, establishing that WAC is required in neurons for normal cognitive performance.\",\n      \"method\": \"Neuronal RNAi knockdown in Drosophila, habituation learning behavioral assay\",\n      \"journal\": \"European journal of human genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean neuronal-specific knockdown with defined behavioral phenotype in a well-established learning paradigm, single lab\",\n      \"pmids\": [\"26757981\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"WAC protein localizes predominantly in the nucleus of cortical neurons after birth and in hippocampal neurons, but is found in the perinuclear region at earlier developmental stages. WAC is also detected in axons and dendrites in a time-dependent manner in primary cultured hippocampal neurons. Nuclear localization is enriched postnatally in cerebral cortex, hippocampus, and cerebellum.\",\n      \"method\": \"Immunohistochemistry, Western blotting with stage-specific brain samples, immunofluorescence in primary cultured neurons\",\n      \"journal\": \"Medical molecular morphology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — direct localization experiment across developmental stages, multiple brain regions, independent antibody preparation; no functional consequence directly linked\",\n      \"pmids\": [\"37402055\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"A nuclear localization domain within WAC was identified and experimentally tested; deletion of this domain impacts the cellular distribution of the WAC protein in GABAergic neurons.\",\n      \"method\": \"Domain deletion constructs with fluorescence-based localization assay in cortical GABAergic neurons\",\n      \"journal\": \"Biology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, single method (fluorescence localization), no functional consequence beyond distribution change\",\n      \"pmids\": [\"37106788\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"WAC is a WW domain- and coiled-coil-containing adaptor protein that (1) promotes transcription-coupled histone H2B ubiquitination by binding RNA polymerase II via its WW domain and recruiting the RNF20/RNF40 E3 ligase complex via its coiled-coil region; (2) regulates starvation-induced autophagy by suppressing GM130-mediated Golgi tethering of GABARAP, enabling centrosomal GABARAP delivery to the phagophore to activate ULK kinase; (3) activates Polo-like kinase 1 for timely mitotic entry by bridging Cdk1-phosphorylated WAC with Plk1's polo-box domain and Aurora A; (4) enhances VCIP135 deubiquitinase activity within a p97/p47 Golgi membrane fusion pathway; (5) protects PINK1 from ubiquitin-dependent degradation to promote mitophagy; and (6) modulates mTORC1 activity through nutrient-sensitive interactions with mTOR-mLST8, R2TP, and TELO2.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"WAC is a WW domain- and coiled-coil-containing adaptor protein that couples active transcription to chromatin modification and serves as a multifunctional scaffold across nuclear, Golgi, and autophagy compartments. Its best-characterized role is to drive transcription-coupled histone H2B monoubiquitination: its N-terminal WW domain engages RNA polymerase II while its C-terminal coiled-coil region binds and stimulates the RNF20/RNF40 E3 ligase, targeting the ligase to active transcription sites, and WAC loss abolishes H2B ubiquitination [#0]. Structural modeling of the human RNF20/RNF40-WAC interface, validated by mutagenesis, defines electrostatic contacts that encode binding specificity essential for H2BUb1 catalysis [#10]. This activity has physiological consequences: WAC is required for plasma cell differentiation and antibody responses through global H2B ubiquitination [#8], sustains MLL-rearranged leukaemic cells [#6], and controls cartilage degradation by linking H2BK120ub1 to H3K27me3 via regulation of KDM6B nuclear entry [#11]. Independently of transcription, WAC regulates starvation-induced autophagy by suppressing GM130-mediated Golgi tethering of GABARAP, releasing a pericentriolar GABARAP pool to the phagophore to activate ULK kinase [#1, #2], and at the Golgi it binds the deubiquitinase VCIP135 and enhances its activity within the p97/p47 membrane fusion pathway [#3]. WAC additionally promotes timely mitotic entry by bridging Cdk1-phosphorylated WAC to the Plk1 polo-box domain and Aurora A to activate Plk1 [#4], protects PINK1 from ubiquitin-dependent degradation to promote mitophagy [#7], and modulates mTORC1 through nutrient-sensitive interactions with mTOR-mLST8, R2TP, and TELO2 [#9]. Neuronal WAC is required for normal habituation learning [#12].\",\n  \"teleology\": [\n    {\n      \"year\": 2002,\n      \"claim\": \"An initial question was what cellular compartment and process this WW-domain adaptor belongs to; nuclear speckle colocalization first hinted at an RNA-processing connection.\",\n      \"evidence\": \"Immunofluorescence colocalization with splicing factor SC35 and phosphorylation detection\",\n      \"pmids\": [\"11827461\"],\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"colocalization only, no functional consequence demonstrated\", \"splicing role never mechanistically established in later work\", \"tyrosine-phosphorylation significance unresolved\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"The central mechanistic role was established by showing WAC bridges RNA polymerase II to the RNF20/RNF40 ligase, defining it as the adaptor that couples transcription to H2B ubiquitination.\",\n      \"evidence\": \"Affinity purification, Co-IP, domain mutational analysis and siRNA depletion with H2B ubiquitination readout\",\n      \"pmids\": [\"21329877\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"structural basis of the RNF20/40 interface not resolved at this stage\", \"downstream transcriptional targets undefined\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"A distinct cytoplasmic function was revealed at the Golgi, where WAC was shown to enhance VCIP135 deubiquitinase activity and is selectively required for p97/p47-mediated Golgi membrane fusion.\",\n      \"evidence\": \"Co-IP, in vitro Golgi reformation and deubiquitinase activity assays, immunofluorescence\",\n      \"pmids\": [\"21811234\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"how nuclear and Golgi pools are partitioned unknown\", \"mechanism by which WAC stimulates VCIP135 unresolved\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"A genome-wide screen established WAC as a positive regulator of starvation-induced autophagosome formation, broadening its role beyond transcription.\",\n      \"evidence\": \"Genome-wide siRNA screen with GFP-LC3 reporter and knockdown validation\",\n      \"pmids\": [\"22354037\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"molecular mechanism not resolved in the screen\", \"link between autophagy role and UPS regulation undefined\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"The autophagy mechanism was resolved: WAC suppresses GM130 tethering of GABARAP to free a pericentriolar GABARAP pool for phagophore delivery and ULK activation.\",\n      \"evidence\": \"Co-IP, knockdown, live-cell imaging, fractionation, in vitro ULK kinase assay and LIR mutagenesis\",\n      \"pmids\": [\"26687599\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"signal that triggers WAC-GM130 competition upon starvation unclear\", \"relationship to WAC's nuclear function not addressed\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Neuronal requirement was demonstrated by showing the Drosophila orthologue is needed for habituation learning, linking WAC to cognitive function.\",\n      \"evidence\": \"Neuronal RNAi knockdown in Drosophila with habituation behavioral assay\",\n      \"pmids\": [\"26757981\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"molecular pathway in neurons not defined\", \"which WAC activity underlies the learning phenotype unknown\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"The transcriptional ligase pathway was given disease relevance by showing WAC is required to maintain MLL-rearranged leukaemic cells via H2B ubiquitination.\",\n      \"evidence\": \"siRNA knockdown with H2B ubiquitination and viability readout plus pharmacological inhibition\",\n      \"pmids\": [\"28690313\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"specific target genes driving leukaemic dependency undefined\", \"selectivity over normal cells unaddressed\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"A cell-cycle function was established: Cdk1-phosphorylated WAC binds the Plk1 polo-box domain and Aurora A to activate Plk1 for timely mitotic entry.\",\n      \"evidence\": \"Co-IP, in vitro kinase assay, Plk1-binding-deficient mutant and rescue with mitotic entry readout\",\n      \"pmids\": [\"30021153\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"how this mitotic role integrates with WAC's chromatin function unclear\", \"structural basis of WAC-PBD interaction not solved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"In vivo significance of the H2B ubiquitination role was shown via a B cell-specific knockout that abolishes plasma cell differentiation and reduces global ubH2B.\",\n      \"evidence\": \"B cell-specific conditional knockout mouse, flow cytometry, ubH2B western blot, antibody response assay\",\n      \"pmids\": [\"37171241\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"specific metabolic target genes regulated not fully mapped\", \"whether non-transcriptional WAC roles contribute unaddressed\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Neuronal localization was characterized as developmentally dynamic, predominantly nuclear postnatally and perinuclear/axodendritic earlier, with a defined nuclear localization domain.\",\n      \"evidence\": \"Immunohistochemistry, Western blotting across brain stages, and domain-deletion fluorescence assays in neurons\",\n      \"pmids\": [\"37402055\", \"37106788\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"functional consequence of subcellular redistribution not established\", \"regulation of the nuclear localization domain unknown\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"A mitophagy function was identified: WAC binds PINK1 transmembrane domains and blocks K137 ubiquitination, stabilizing PINK1 to promote mitophagy and osteogenic differentiation.\",\n      \"evidence\": \"Co-IP, ubiquitination assay, K137 site mutagenesis, in vitro and in vivo bone formation assays\",\n      \"pmids\": [\"39555688\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"E3 ligase WAC competes with not identified\", \"relationship to GABARAP-dependent autophagy role unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"A nutrient-sensing function was proposed by showing direct WAC binding to mTOR-mLST8, R2TP, and TELO2 in nutrient-modulated complexes correlating with mTORC1 activity.\",\n      \"evidence\": \"In vitro binding with purified proteins and Co-IP under defined nutrient conditions plus omics\",\n      \"pmids\": [\"40653822\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"causal effect of WAC on mTORC1 output not demonstrated by loss-of-function\", \"structural basis of binding unresolved\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"The epigenetic crosstalk role was extended by showing WAC controls H2BK120ub1 and influences H3K27me3 via KDM6B nuclear entry, with cartilage-specific knockout protecting against arthritis.\",\n      \"evidence\": \"Cartilage-specific conditional KO, ChIP for histone marks, nuclear fractionation for KDM6B, in vivo arthritis models\",\n      \"pmids\": [\"40893665\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"mechanism by which WAC regulates KDM6B import undefined\", \"direct gene targets of the ub-methyl crosstalk not enumerated\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"The structural basis of the core adaptor function was defined, mapping the RNF20/RNF40-WAC interface and its electrostatic specificity determinants required for H2BUb1 catalysis.\",\n      \"evidence\": \"X-ray crystallography of yeast Bre1-Lge1, AlphaFold modeling of human RNF20/40-WAC, and mutagenesis with functional validation\",\n      \"pmids\": [\"41533567\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"full-length human complex not crystallized\", \"how WW-domain Pol II engagement couples to ligase geometry unresolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unknown how WAC's distinct nuclear (H2B ubiquitination), Golgi/autophagy, mitotic, and nutrient-sensing activities are coordinated within a single cell and which are regulated by the same post-translational or localization switches.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"no unifying regulatory model integrating compartment-specific functions\", \"signals partitioning WAC between nucleus, Golgi, and centrosome undefined\", \"in vivo loss-of-function for mTORC1 and mitophagy roles lacking\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1, 3, 4]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 3, 4, 7]},\n      {\"term_id\": \"GO:0042393\", \"supporting_discovery_ids\": [0, 10]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 3, 13]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [1, 3]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [0, 10, 11]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [0, 8]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [1, 2, 7]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"complexes\": [\n      \"RNF20/RNF40-WAC H2B ubiquitination complex\",\n      \"p97/p47-VCIP135 Golgi membrane fusion complex\"\n    ],\n    \"partners\": [\n      \"RNF20\",\n      \"RNF40\",\n      \"GM130\",\n      \"GABARAP\",\n      \"VCIP135\",\n      \"PLK1\",\n      \"PINK1\",\n      \"MTOR\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}