{"gene":"KANSL3","run_date":"2026-06-10T01:55:23","timeline":{"discoveries":[{"year":2010,"finding":"KANSL3 (NSL3) is a subunit of the NSL complex (NSL1, NSL2, NSL3, MCRS2, MBD-R2, WDS) that associates with the histone acetyltransferase MOF in both Drosophila and mammals. Depletion of NSL3 severely affects gene expression genome-wide. Tethering of NSL3 to a heterologous promoter leads to robust transcription activation dependent on NSL1, MCRS2, and MOF levels. NSL complex members bind target promoters independently of MOF, but depletion of MCRS2 affects MOF recruitment.","method":"Biochemical purification, ChIP-Seq, RNAi depletion, heterologous promoter tethering assay","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal biochemical characterization, genome-wide ChIP-Seq, functional depletion, and tethering assay; replicated across Drosophila and mammals","pmids":["20620954"],"is_preprint":false},{"year":2012,"finding":"NSL3 (KANSL3) binds promoters of housekeeping genes in Drosophila and is required for efficient recruitment of RNA Polymerase II to NSL target gene promoters. NSL3 depletion reduces TBP and TFIIB binding at target promoters, indicating the NSL complex is required for optimal pre-initiation complex recruitment. NSL-bound promoters are associated with H4K16ac, H3K4me2, H3K4me3, and H3K9ac histone modifications.","method":"ChIP-seq (NSL1, NSL3, MBD-R2, MCRS2), RNA Pol II ChIP-seq in NSL3-depleted cells, RNAi knockdown","journal":"PLoS genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — genome-wide ChIP-seq with functional depletion and multiple orthogonal chromatin readouts; replicated across multiple NSL subunits","pmids":["22723752"],"is_preprint":false},{"year":2015,"finding":"During mitosis, KANSL3 relocalizes from chromatin to the mitotic spindle in a RanGTP-dependent manner. KANSL3 is identified as a microtubule minus-end-binding protein that stabilizes microtubule minus ends, and is essential for spindle assembly and chromosome segregation.","method":"Live-cell imaging, immunofluorescence localization, RanGTP-dependent spindle assembly assay, loss-of-function with mitotic phenotype readout","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — direct localization experiments with functional consequence, biochemical spindle assays, and RanGTP dependence demonstrated with multiple orthogonal approaches","pmids":["26243146"],"is_preprint":false},{"year":2016,"finding":"MOF and a subset of NSL complex partners, including KANSL3, reside in mitochondria. MOF binds mtDNA, and this binding is dependent on KANSL3. MOF regulates oxidative phosphorylation by controlling expression of respiratory genes from both nuclear and mtDNA.","method":"Subcellular fractionation, immunofluorescence, mtDNA ChIP, conditional knockout mouse model with cardiac phenotype","journal":"Cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — fractionation and ChIP establish mitochondrial localization and KANSL3-dependent mtDNA binding; in vivo mouse KO validates functional consequence","pmids":["27768893"],"is_preprint":false},{"year":2017,"finding":"OGT1 physically interacts with NSL3 (KANSL3) and O-GlcNAcylates it, stabilizing NSL3 protein. This stabilization promotes NSL complex histone acetyltransferase activity, leading to increased global acetylation of histone H4 at K5, K8, and K16. Knockdown or overexpression of OGT1 markedly affects these H4 acetylation levels.","method":"Co-immunoprecipitation, in vitro O-GlcNAc transferase assay, wheat germ agglutinin affinity purification, OGT1 catalytic mutant (C964A) co-transfection","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — in vitro transferase assay combined with Co-IP, affinity purification, and catalytic mutant, multiple orthogonal methods in a single study","pmids":["28450392"],"is_preprint":false},{"year":2019,"finding":"O-GlcNAcylation of KANSL3 at Thr755 by OGT1 is required for NSL complex integrity and holoenzyme activity. Mutation T755A promotes ubiquitin-mediated proteasomal degradation of NSL3. UBE2S (ubiquitin-conjugating enzyme E2 S) directly binds NSL3 and accelerates its degradation. OGT1 and UBE2S competitively bind to NSL3, coordinating its stability.","method":"In vitro O-GlcNAc transferase assay combined with mass spectrometry, site-directed mutagenesis (T755A), co-immunoprecipitation, ubiquitination assay","journal":"International journal of molecular sciences","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro transferase assay with mass spectrometry site identification, mutagenesis, and competitive binding Co-IP in one study","pmids":["31881804"],"is_preprint":false},{"year":2019,"finding":"In Drosophila S2 cells, depletion of Rcd1 (KANSL3 ortholog) by RNAi leads to defects in chromosome segregation, reduced levels of centromere component Cid/CENP-A and kinetochore component Ndc80, and negatively affects centriole duplication. Rcd1-GFP accumulates at centrosomes and the telophase midbody during mitosis. RT-qPCR showed that transcription of centromere/kinetochore genes (cid, Mis12, Nnf1b) and centriole duplication genes is substantially reduced in Rcd1 RNAi cells, suggesting mitotic phenotypes are primarily due to transcriptional downregulation of these genes.","method":"RNAi depletion, immunofluorescence, GFP live-cell localization, RT-qPCR","journal":"PLoS genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RNAi with defined cellular phenotype and GFP localization; mechanism attributed primarily to transcriptional effects; single study","pmids":["31527906"],"is_preprint":false},{"year":2019,"finding":"NSL3 (KANSL3) binds to TATA-less promoters in a sequence-dependent manner. The NSL complex interacts with the NURF chromatin remodeling complex and is necessary and sufficient to recruit NURF to target promoters, thereby maintaining nucleosome-depleted regions at transcription start sites and enabling accurate TSS selection.","method":"ChIP-seq, NSL1 depletion with nucleosome positioning analysis, biochemical interaction with NURF complex","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 2 / Strong — ChIP-seq, nucleosome positioning, biochemical NURF interaction, and rescue experiments across multiple orthogonal approaches","pmids":["30819819"],"is_preprint":false},{"year":2020,"finding":"Neural-specific depletion of Kansl3 (along with Mof or Kansl2) causes widespread metabolic defects including accumulation of free long-chain fatty acids (LCFAs). LCFAs trigger TLR4-NFκB-dependent pro-inflammatory signaling in neighboring vascular pericytes, leading to pericyte activation and vascular breakdown. This establishes KANSL3 as part of a pathway linking epigenetic regulation to neurovascular homeostasis via metabolic intermediates.","method":"Conditional neural-specific knockout in mice, metabolomics, TLR4 inhibitor rescue, immunofluorescence of pericyte markers","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional KO in mice with defined molecular pathway (LCFA → TLR4-NFκB), metabolomic readout, and pharmacological rescue","pmids":["32541879"],"is_preprint":false},{"year":2022,"finding":"NSL3 (KANSL3) knockout in human 293T cells, identified by CRISPR/Cas9 and ChIP-seq, reveals over 100 transcriptional targets including YY1. MOF and NSL3 co-localize with H4K16ac, H3K4me2, and H3K4me3 at the YY1 TSS. NSL3 silencing reduces YY1 expression; NSL3 knockout suppresses CDC6 (a YY1 target) expression. NSL3 regulates clonogenic ability of HepG2 cells, which is rescued by YY1 overexpression, placing YY1 downstream of KANSL3.","method":"CRISPR/Cas9 NSL3-KO, ChIP-seq, siRNA knockdown, overexpression, colony formation assay","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CRISPR KO with ChIP-seq and epistasis rescue in a single lab study","pmids":["35409160"],"is_preprint":false},{"year":2022,"finding":"NSL3 (KANSL3) knockout promotes cell invasion in cancer cells and positively correlates with mesenchymal biomarkers (N-cadherin, vimentin, snail). NSL3-KO causes lumen-like cell morphology. ChIP-seq indicates NSL complex may be involved in phosphoinositide-mediated signaling pathways. Unlike MSL1, NSL3 does not bind the E-box-containing Snail promoter.","method":"CRISPR/Cas9 KO, Transwell invasion assay, immunostaining, ChIP-seq","journal":"Cellular and molecular life sciences : CMLS","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CRISPR KO with defined cellular phenotype and ChIP-seq, but single lab study","pmids":["35416545"],"is_preprint":false},{"year":2023,"finding":"Germline-specific knockdown of NSL3 (KANSL3) in Drosophila results in reduced piRNA production from a subset of bidirectional piRNA clusters, with widespread transposon derepression. This establishes a role for the NSL complex in promoting transcription of piRNA precursors from telomeric piRNA clusters.","method":"RNAi germline-specific knockdown, small RNA sequencing, ChIP-seq","journal":"Life science alliance","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RNAi with genome-wide sequencing readouts in a single study; NSL3 effect described alongside NSL1 and NSL2","pmids":["37399316"],"is_preprint":false},{"year":2023,"finding":"Deletion of KANSL3 in postmitotic podocytes leads to catastrophic kidney dysfunction. KANSL3 ablation disrupts microtubule dynamics and podocyte functions in nonciliated cells, while in ciliated fibroblasts it leads to loss of cilia and impaired sonic hedgehog pathway. The NSL complex is a master regulator of intraciliary transport genes in both dividing and nondividing cells.","method":"Kidney-specific conditional knockout, single-cell RNA sequencing, cilia immunofluorescence, microtubule dynamics assay","journal":"Science advances","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional KO in vivo with multiple defined cellular phenotypes and mechanistic pathway analysis (IFT, Shh, microtubule dynamics)","pmids":["37624894"],"is_preprint":false},{"year":2024,"finding":"Homozygous knockout of Kansl3 in mice leads to embryonic lethality at peri-implantation stages; embryos fail to hatch from the zona pellucida. Kansl3-null embryos have a significantly reduced inner cell mass (ICM) cell number with normal trophectoderm cell numbers, and both epiblast and primitive endoderm lineages show reduced cell numbers.","method":"Conditional/homozygous KO mouse, zona pellucida removal in vitro, immunofluorescence lineage marker analysis","journal":"Molecular reproduction and development","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KO with defined lineage-specific cellular phenotype, single study","pmids":["38769918"],"is_preprint":false},{"year":2025,"finding":"TRAP1 (molecular chaperone) directly interacts with KANSL3, as demonstrated by co-immunoprecipitation and LC-MS/MS. Under diabetic/high glucose-palmitate conditions, TRAP1-KANSL3 interaction decreases, and KANSL3 acetylation increases. TRAP1 inhibits KANSL3 acetylation under normal conditions to preserve mitophagy; loss of TRAP1 under diabetic conditions impairs mitophagy and mitochondrial function.","method":"Co-immunoprecipitation, LC-MS/MS, lentiviral TRAP1 overexpression and KANSL3 knockdown, mitophagy flux assay (mKeima), transmission electron microscopy","journal":"Cell communication and signaling : CCS","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP with MS confirmation of interaction, functional rescue experiments; single lab study","pmids":["41039555"],"is_preprint":false},{"year":2025,"finding":"Hepatocyte-specific deletion of KANSL3 in mice results in early-onset liver disease marked by biliary hyperplasia and hepatic fibrosis. KANSL3 regulates hepatocyte transcriptional networks for hepatic steroid and lipid metabolism through histone acetylation. Loss of KANSL3 disrupts hepatocyte differentiation in vivo and impairs transcriptional programs for hepatocyte differentiation in ductal and fetal liver organoids.","method":"Hepatocyte-specific conditional KO, single-cell RNA sequencing, organoid differentiation assay, histone acetylation analysis","journal":"Life science alliance","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional KO in vivo with scRNA-seq, organoid functional rescue assays, and histone acetylation readouts across multiple orthogonal methods","pmids":["41044006"],"is_preprint":false}],"current_model":"KANSL3 is a core subunit of the evolutionarily conserved NSL histone acetyltransferase complex that associates with MOF/KAT8; during interphase it binds to gene promoters in a sequence-dependent manner to recruit the NURF remodeling complex, maintain nucleosome-depleted regions, and activate transcription of thousands of housekeeping and tissue-specific genes via H4K5/K8/K16 acetylation, while during mitosis it relocalizes to the spindle as a microtubule minus-end-binding and stabilizing factor essential for chromosome segregation; its protein stability is regulated by OGT1-mediated O-GlcNAcylation at Thr755 (opposing UBE2S-mediated ubiquitin degradation) and by TRAP1-mediated inhibition of its acetylation; loss of KANSL3 in vivo causes tissue-specific phenotypes including peri-implantation lethality, neurovascular breakdown through LCFA-TLR4-NFκB signaling, kidney dysfunction with impaired intraciliary transport, and liver disease through defective hepatocyte differentiation and lipid metabolism."},"narrative":{"mechanistic_narrative":"KANSL3 is a core subunit of the evolutionarily conserved NSL histone acetyltransferase complex that associates with the acetyltransferase MOF and other NSL members to activate transcription genome-wide [PMID:20620954]. At target promoters—predominantly TATA-less, housekeeping-gene promoters bound in a sequence-dependent manner—the complex maintains nucleosome-depleted regions and ensures accurate transcription start-site selection by recruiting the NURF chromatin remodeler, and is required for efficient assembly of the RNA Polymerase II pre-initiation complex (TBP, TFIIB) coincident with active chromatin marks including H4K16ac, H3K4me2/3, and H3K9ac [PMID:22723752, PMID:30819819]. KANSL3 carries out a moonlighting structural role outside chromatin: during mitosis it relocalizes to the spindle in a RanGTP-dependent manner, where it binds and stabilizes microtubule minus ends to enable spindle assembly and chromosome segregation [PMID:26243146]. KANSL3 also partitions to mitochondria, where it is required for MOF binding to mtDNA and the consequent expression of respiratory genes [PMID:27768893]. Its abundance and activity are post-translationally controlled: OGT1-mediated O-GlcNAcylation at Thr755 stabilizes KANSL3 and sustains NSL complex integrity and H4K5/K8/K16 acetylation, opposing UBE2S-driven ubiquitin-proteasomal degradation, with OGT1 and UBE2S competing for binding [PMID:28450392, PMID:31881804], while TRAP1 binding restrains KANSL3 acetylation to preserve mitophagy under metabolic stress [PMID:41039555]. Through these transcriptional and structural roles, KANSL3 loss in vivo produces tissue-specific pathology—peri-implantation lethality with reduced inner cell mass [PMID:38769918], neurovascular breakdown via LCFA-driven TLR4-NFκB signaling [PMID:32541879], kidney dysfunction through disrupted intraciliary-transport gene programs and microtubule dynamics [PMID:37624894], and liver disease through defective hepatocyte differentiation and lipid metabolism [PMID:41044006].","teleology":[{"year":2010,"claim":"Established that KANSL3 is a bona fide subunit of the MOF-associated NSL complex and a positive regulator of genome-wide transcription, answering whether KANSL3 functions in chromatin-based gene activation.","evidence":"Biochemical purification, ChIP-Seq, RNAi depletion, and heterologous promoter tethering in Drosophila and mammals","pmids":["20620954"],"confidence":"High","gaps":["Did not define which promoter features dictate NSL targeting","Direct enzymatic contribution of KANSL3 to acetylation not isolated from MOF"]},{"year":2012,"claim":"Showed the NSL complex acts at the pre-initiation step by promoting RNA Pol II, TBP, and TFIIB recruitment at housekeeping-gene promoters, defining the transcriptional mechanism.","evidence":"ChIP-seq of NSL subunits and Pol II in NSL3-depleted Drosophila cells with RNAi knockdown","pmids":["22723752"],"confidence":"High","gaps":["Mechanism of PIC stabilization not resolved","Did not address non-chromatin roles"]},{"year":2015,"claim":"Revealed a non-transcriptional structural function: KANSL3 relocalizes to the mitotic spindle and acts as a microtubule minus-end-binding stabilizer essential for chromosome segregation.","evidence":"Live-cell imaging, immunofluorescence, RanGTP-dependent spindle assembly assay, and loss-of-function mitotic readouts","pmids":["26243146"],"confidence":"High","gaps":["Structural basis of minus-end binding not defined","Relationship between chromatin and spindle pools unclear"]},{"year":2016,"claim":"Demonstrated a mitochondrial pool of KANSL3 required for MOF–mtDNA association and respiratory gene expression, extending NSL function to oxidative phosphorylation.","evidence":"Subcellular fractionation, mtDNA ChIP, and conditional cardiac knockout mouse","pmids":["27768893"],"confidence":"High","gaps":["Direct mtDNA-binding activity of KANSL3 itself not shown","How nuclear vs mitochondrial partitioning is controlled is unknown"]},{"year":2017,"claim":"Identified OGT1-mediated O-GlcNAcylation as a stabilizing modification of KANSL3 that boosts NSL acetyltransferase output on H4K5/K8/K16.","evidence":"Co-IP, in vitro O-GlcNAc transferase assay, WGA affinity purification, and OGT1 catalytic mutant in cells","pmids":["28450392"],"confidence":"High","gaps":["Modified residue not yet mapped at this stage","In vivo physiological trigger of O-GlcNAcylation unknown"]},{"year":2019,"claim":"Mapped the stabilizing O-GlcNAc site to Thr755 and showed OGT1 and UBE2S compete to set KANSL3 stability and NSL holoenzyme integrity, defining the degradation control axis.","evidence":"In vitro transferase assay with mass spectrometry, T755A mutagenesis, Co-IP, and ubiquitination assay","pmids":["31881804"],"confidence":"High","gaps":["UBE2S substrate-recognition determinants not defined","Upstream signals shifting the OGT1/UBE2S balance unknown"]},{"year":2019,"claim":"Defined how NSL achieves nucleosome-free promoters and accurate TSS selection: KANSL3 binds TATA-less promoters sequence-dependently and recruits NURF to maintain nucleosome-depleted regions.","evidence":"ChIP-seq, nucleosome positioning after NSL1 depletion, and biochemical NSL–NURF interaction","pmids":["30819819"],"confidence":"High","gaps":["Sequence motif recognized by KANSL3 not specified","Domain mediating NURF contact not mapped"]},{"year":2019,"claim":"Indicated that mitotic defects after KANSL3-ortholog loss in Drosophila arise largely from transcriptional downregulation of centromere/kinetochore and centriole genes, linking transcriptional and mitotic phenotypes.","evidence":"RNAi depletion of Rcd1, immunofluorescence, GFP localization, and RT-qPCR in S2 cells","pmids":["31527906"],"confidence":"Medium","gaps":["Single study; transcriptional vs direct spindle contributions not fully separated","Centrosome/midbody localization function not mechanistically dissected"]},{"year":2020,"claim":"Connected KANSL3-dependent epigenetic regulation to organ physiology by showing neural loss causes LCFA accumulation that drives TLR4-NFκB pericyte activation and neurovascular breakdown.","evidence":"Neural-specific conditional KO mice, metabolomics, TLR4 inhibitor rescue, and pericyte marker immunofluorescence","pmids":["32541879"],"confidence":"High","gaps":["Direct transcriptional targets governing LCFA metabolism not enumerated","Cell-autonomous vs paracrine steps only partly separated"]},{"year":2022,"claim":"Identified YY1 as a key downstream transcriptional target in human cells, with KANSL3-driven clonogenicity rescued by YY1, placing YY1 epistatically below KANSL3.","evidence":"CRISPR/Cas9 KO, ChIP-seq, siRNA, overexpression, and colony formation in 293T/HepG2 cells","pmids":["35409160"],"confidence":"Medium","gaps":["Single-lab study","Generalizability of YY1 axis to other cell types untested"]},{"year":2022,"claim":"Showed KANSL3 loss promotes mesenchymal/invasive phenotypes, implicating NSL transcriptional regulation in epithelial cell identity.","evidence":"CRISPR/Cas9 KO, Transwell invasion assay, immunostaining, and ChIP-seq","pmids":["35416545"],"confidence":"Medium","gaps":["Direct vs indirect control of mesenchymal markers unresolved","Phosphoinositide-signaling link only correlative"]},{"year":2023,"claim":"Extended NSL transcriptional function to small-RNA biology, showing KANSL3 promotes transcription of piRNA precursors and restrains transposons in the Drosophila germline.","evidence":"Germline-specific RNAi, small RNA sequencing, and ChIP-seq","pmids":["37399316"],"confidence":"Medium","gaps":["Single study with effect described alongside other NSL subunits","Conservation of piRNA role in mammals untested"]},{"year":2023,"claim":"Established the NSL complex as a master regulator of intraciliary-transport genes and microtubule dynamics in both dividing and postmitotic cells, explaining kidney pathology on KANSL3 loss.","evidence":"Kidney-specific conditional KO, single-cell RNA-seq, cilia immunofluorescence, and microtubule dynamics assays","pmids":["37624894"],"confidence":"High","gaps":["Whether ciliary defect is purely transcriptional or also reflects direct microtubule role not fully separated","Specific IFT target genes driving phenotype not pinpointed"]},{"year":2024,"claim":"Defined the earliest developmental requirement, showing Kansl3 is essential at peri-implantation with selective reduction of inner cell mass lineages.","evidence":"Homozygous KO mice, in vitro zona pellucida removal, and lineage-marker immunofluorescence","pmids":["38769918"],"confidence":"Medium","gaps":["Molecular targets underlying ICM-specific defect not identified","Single study"]},{"year":2025,"claim":"Identified TRAP1 as a direct KANSL3 partner that restrains its acetylation to preserve mitophagy, adding an acetylation-based control layer relevant to metabolic stress.","evidence":"Co-IP, LC-MS/MS, TRAP1 overexpression and KANSL3 knockdown, mKeima mitophagy flux, and TEM","pmids":["41039555"],"confidence":"Medium","gaps":["Acetyltransferase responsible for KANSL3 acetylation not identified","Single-lab study; in vivo relevance untested"]},{"year":2025,"claim":"Demonstrated that KANSL3 governs hepatocyte differentiation and lipid/steroid metabolic transcription, with loss causing biliary hyperplasia and fibrosis.","evidence":"Hepatocyte-specific conditional KO, single-cell RNA-seq, organoid differentiation, and histone acetylation analysis","pmids":["41044006"],"confidence":"High","gaps":["Key direct target genes of hepatocyte program not isolated","Whether metabolic phenotype is cell-autonomous to hepatocytes not fully resolved"]},{"year":null,"claim":"How KANSL3 dynamically partitions among nuclear chromatin, mitotic spindle, and mitochondria, and what governs its tissue-specific target gene selection, remain open.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of KANSL3 domains for DNA, microtubule, and partner binding","Mechanism of context-dependent localization switching unknown","Sequence determinants of promoter targeting unmapped"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[7]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[0,1,7]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[2]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,4]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,1,7]},{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[3]}],"pathway":[{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[0,1,7]},{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[7]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[2]}],"complexes":["NSL (KANSL/MOF) histone acetyltransferase complex"],"partners":["KAT8","OGT1","UBE2S","TRAP1","NURF"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9P2N6","full_name":"KAT8 regulatory NSL complex subunit 3","aliases":["NSL complex protein NSL3","Non-specific lethal 3 homolog","Serum inhibited-related protein","Testis development protein PRTD"],"length_aa":904,"mass_kda":96.0,"function":"Non-catalytic component of the NSL histone acetyltransferase complex, a multiprotein complex that mediates histone H4 acetylation at 'Lys-5'- and 'Lys-8' (H4K5ac and H4K8ac) at transcription start sites and promotes transcription initiation (PubMed:20018852, PubMed:33657400). The NSL complex also acts as a regulator of gene expression in mitochondria (PubMed:27768893). Within the NSL complex, KANSL3 is required to promote KAT8 association with mitochondrial DNA (PubMed:27768893). Required for transcription of intraciliary transport genes in both ciliated and non-ciliated cells (By similarity). This is necessary for cilium assembly in ciliated cells and for organization of the microtubule cytoskeleton in non-ciliated cells (By similarity). Also required within the NSL complex to maintain nuclear architecture stability by promoting KAT8-mediated acetylation of lamin LMNA (By similarity). Plays an essential role in spindle assembly during mitosis (PubMed:26243146). Acts as a microtubule minus-end binding protein which stabilizes microtubules and promotes their assembly (PubMed:26243146). Indispensable during early embryonic development where it is required for proper lineage specification and maintenance during peri-implantation development and is essential for implantation (By similarity)","subcellular_location":"Nucleus; Mitochondrion; Cytoplasm, cytoskeleton, spindle pole","url":"https://www.uniprot.org/uniprotkb/Q9P2N6/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/KANSL3","classification":"Common Essential","n_dependent_lines":1179,"n_total_lines":1208,"dependency_fraction":0.9759933774834437},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/KANSL3","total_profiled":1310},"omim":[{"mim_id":"617742","title":"KAT8 REGULATORY NSL COMPLEX, SUBUNIT 3; KANSL3","url":"https://www.omim.org/entry/617742"},{"mim_id":"615488","title":"KAT8 REGULATORY NSL COMPLEX, SUBUNIT 2; KANSL2","url":"https://www.omim.org/entry/615488"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"},{"location":"Vesicles","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/KANSL3"},"hgnc":{"alias_symbol":["FLJ10081","Rcd1","NSL3"],"prev_symbol":["KIAA1310"]},"alphafold":{"accession":"Q9P2N6","domains":[{"cath_id":"-","chopping":"92-254","consensus_level":"high","plddt":86.9196,"start":92,"end":254},{"cath_id":"3.40.50.1820","chopping":"258-475","consensus_level":"high","plddt":89.5065,"start":258,"end":475}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9P2N6","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9P2N6-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9P2N6-F1-predicted_aligned_error_v6.png","plddt_mean":60.12},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=KANSL3","jax_strain_url":"https://www.jax.org/strain/search?query=KANSL3"},"sequence":{"accession":"Q9P2N6","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9P2N6.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9P2N6/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9P2N6"}},"corpus_meta":[{"pmid":"27768893","id":"PMC_27768893","title":"MOF Acetyl Transferase Regulates Transcription and Respiration in Mitochondria.","date":"2016","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/27768893","citation_count":150,"is_preprint":false},{"pmid":"20620954","id":"PMC_20620954","title":"The nonspecific lethal complex is a transcriptional regulator in Drosophila.","date":"2010","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/20620954","citation_count":121,"is_preprint":false},{"pmid":"22723752","id":"PMC_22723752","title":"The NSL complex regulates housekeeping genes in Drosophila.","date":"2012","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/22723752","citation_count":81,"is_preprint":false},{"pmid":"26243146","id":"PMC_26243146","title":"An epigenetic regulator emerges as microtubule minus-end binding and stabilizing factor in mitosis.","date":"2015","source":"Nature 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disease","url":"https://pubmed.ncbi.nlm.nih.gov/35973998","citation_count":14,"is_preprint":false},{"pmid":"31527906","id":"PMC_31527906","title":"RNAi-mediated depletion of the NSL complex subunits leads to abnormal chromosome segregation and defective centrosome duplication in Drosophila mitosis.","date":"2019","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/31527906","citation_count":12,"is_preprint":false},{"pmid":"30819819","id":"PMC_30819819","title":"The NSL complex-mediated nucleosome landscape is required to maintain transcription fidelity and suppression of transcription noise.","date":"2019","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/30819819","citation_count":11,"is_preprint":false},{"pmid":"35409160","id":"PMC_35409160","title":"The Non-Specific Lethal (NSL) Histone Acetyltransferase Complex Transcriptionally Regulates Yin Yang 1-Mediated Cell Proliferation in Human Cells.","date":"2022","source":"International journal of 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Depletion of NSL3 severely affects gene expression genome-wide. Tethering of NSL3 to a heterologous promoter leads to robust transcription activation dependent on NSL1, MCRS2, and MOF levels. NSL complex members bind target promoters independently of MOF, but depletion of MCRS2 affects MOF recruitment.\",\n      \"method\": \"Biochemical purification, ChIP-Seq, RNAi depletion, heterologous promoter tethering assay\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal biochemical characterization, genome-wide ChIP-Seq, functional depletion, and tethering assay; replicated across Drosophila and mammals\",\n      \"pmids\": [\"20620954\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"NSL3 (KANSL3) binds promoters of housekeeping genes in Drosophila and is required for efficient recruitment of RNA Polymerase II to NSL target gene promoters. NSL3 depletion reduces TBP and TFIIB binding at target promoters, indicating the NSL complex is required for optimal pre-initiation complex recruitment. NSL-bound promoters are associated with H4K16ac, H3K4me2, H3K4me3, and H3K9ac histone modifications.\",\n      \"method\": \"ChIP-seq (NSL1, NSL3, MBD-R2, MCRS2), RNA Pol II ChIP-seq in NSL3-depleted cells, RNAi knockdown\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genome-wide ChIP-seq with functional depletion and multiple orthogonal chromatin readouts; replicated across multiple NSL subunits\",\n      \"pmids\": [\"22723752\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"During mitosis, KANSL3 relocalizes from chromatin to the mitotic spindle in a RanGTP-dependent manner. KANSL3 is identified as a microtubule minus-end-binding protein that stabilizes microtubule minus ends, and is essential for spindle assembly and chromosome segregation.\",\n      \"method\": \"Live-cell imaging, immunofluorescence localization, RanGTP-dependent spindle assembly assay, loss-of-function with mitotic phenotype readout\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — direct localization experiments with functional consequence, biochemical spindle assays, and RanGTP dependence demonstrated with multiple orthogonal approaches\",\n      \"pmids\": [\"26243146\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"MOF and a subset of NSL complex partners, including KANSL3, reside in mitochondria. MOF binds mtDNA, and this binding is dependent on KANSL3. MOF regulates oxidative phosphorylation by controlling expression of respiratory genes from both nuclear and mtDNA.\",\n      \"method\": \"Subcellular fractionation, immunofluorescence, mtDNA ChIP, conditional knockout mouse model with cardiac phenotype\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — fractionation and ChIP establish mitochondrial localization and KANSL3-dependent mtDNA binding; in vivo mouse KO validates functional consequence\",\n      \"pmids\": [\"27768893\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"OGT1 physically interacts with NSL3 (KANSL3) and O-GlcNAcylates it, stabilizing NSL3 protein. This stabilization promotes NSL complex histone acetyltransferase activity, leading to increased global acetylation of histone H4 at K5, K8, and K16. Knockdown or overexpression of OGT1 markedly affects these H4 acetylation levels.\",\n      \"method\": \"Co-immunoprecipitation, in vitro O-GlcNAc transferase assay, wheat germ agglutinin affinity purification, OGT1 catalytic mutant (C964A) co-transfection\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — in vitro transferase assay combined with Co-IP, affinity purification, and catalytic mutant, multiple orthogonal methods in a single study\",\n      \"pmids\": [\"28450392\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"O-GlcNAcylation of KANSL3 at Thr755 by OGT1 is required for NSL complex integrity and holoenzyme activity. Mutation T755A promotes ubiquitin-mediated proteasomal degradation of NSL3. UBE2S (ubiquitin-conjugating enzyme E2 S) directly binds NSL3 and accelerates its degradation. OGT1 and UBE2S competitively bind to NSL3, coordinating its stability.\",\n      \"method\": \"In vitro O-GlcNAc transferase assay combined with mass spectrometry, site-directed mutagenesis (T755A), co-immunoprecipitation, ubiquitination assay\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro transferase assay with mass spectrometry site identification, mutagenesis, and competitive binding Co-IP in one study\",\n      \"pmids\": [\"31881804\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"In Drosophila S2 cells, depletion of Rcd1 (KANSL3 ortholog) by RNAi leads to defects in chromosome segregation, reduced levels of centromere component Cid/CENP-A and kinetochore component Ndc80, and negatively affects centriole duplication. Rcd1-GFP accumulates at centrosomes and the telophase midbody during mitosis. RT-qPCR showed that transcription of centromere/kinetochore genes (cid, Mis12, Nnf1b) and centriole duplication genes is substantially reduced in Rcd1 RNAi cells, suggesting mitotic phenotypes are primarily due to transcriptional downregulation of these genes.\",\n      \"method\": \"RNAi depletion, immunofluorescence, GFP live-cell localization, RT-qPCR\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNAi with defined cellular phenotype and GFP localization; mechanism attributed primarily to transcriptional effects; single study\",\n      \"pmids\": [\"31527906\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"NSL3 (KANSL3) binds to TATA-less promoters in a sequence-dependent manner. The NSL complex interacts with the NURF chromatin remodeling complex and is necessary and sufficient to recruit NURF to target promoters, thereby maintaining nucleosome-depleted regions at transcription start sites and enabling accurate TSS selection.\",\n      \"method\": \"ChIP-seq, NSL1 depletion with nucleosome positioning analysis, biochemical interaction with NURF complex\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — ChIP-seq, nucleosome positioning, biochemical NURF interaction, and rescue experiments across multiple orthogonal approaches\",\n      \"pmids\": [\"30819819\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Neural-specific depletion of Kansl3 (along with Mof or Kansl2) causes widespread metabolic defects including accumulation of free long-chain fatty acids (LCFAs). LCFAs trigger TLR4-NFκB-dependent pro-inflammatory signaling in neighboring vascular pericytes, leading to pericyte activation and vascular breakdown. This establishes KANSL3 as part of a pathway linking epigenetic regulation to neurovascular homeostasis via metabolic intermediates.\",\n      \"method\": \"Conditional neural-specific knockout in mice, metabolomics, TLR4 inhibitor rescue, immunofluorescence of pericyte markers\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional KO in mice with defined molecular pathway (LCFA → TLR4-NFκB), metabolomic readout, and pharmacological rescue\",\n      \"pmids\": [\"32541879\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"NSL3 (KANSL3) knockout in human 293T cells, identified by CRISPR/Cas9 and ChIP-seq, reveals over 100 transcriptional targets including YY1. MOF and NSL3 co-localize with H4K16ac, H3K4me2, and H3K4me3 at the YY1 TSS. NSL3 silencing reduces YY1 expression; NSL3 knockout suppresses CDC6 (a YY1 target) expression. NSL3 regulates clonogenic ability of HepG2 cells, which is rescued by YY1 overexpression, placing YY1 downstream of KANSL3.\",\n      \"method\": \"CRISPR/Cas9 NSL3-KO, ChIP-seq, siRNA knockdown, overexpression, colony formation assay\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CRISPR KO with ChIP-seq and epistasis rescue in a single lab study\",\n      \"pmids\": [\"35409160\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"NSL3 (KANSL3) knockout promotes cell invasion in cancer cells and positively correlates with mesenchymal biomarkers (N-cadherin, vimentin, snail). NSL3-KO causes lumen-like cell morphology. ChIP-seq indicates NSL complex may be involved in phosphoinositide-mediated signaling pathways. Unlike MSL1, NSL3 does not bind the E-box-containing Snail promoter.\",\n      \"method\": \"CRISPR/Cas9 KO, Transwell invasion assay, immunostaining, ChIP-seq\",\n      \"journal\": \"Cellular and molecular life sciences : CMLS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CRISPR KO with defined cellular phenotype and ChIP-seq, but single lab study\",\n      \"pmids\": [\"35416545\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Germline-specific knockdown of NSL3 (KANSL3) in Drosophila results in reduced piRNA production from a subset of bidirectional piRNA clusters, with widespread transposon derepression. This establishes a role for the NSL complex in promoting transcription of piRNA precursors from telomeric piRNA clusters.\",\n      \"method\": \"RNAi germline-specific knockdown, small RNA sequencing, ChIP-seq\",\n      \"journal\": \"Life science alliance\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNAi with genome-wide sequencing readouts in a single study; NSL3 effect described alongside NSL1 and NSL2\",\n      \"pmids\": [\"37399316\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Deletion of KANSL3 in postmitotic podocytes leads to catastrophic kidney dysfunction. KANSL3 ablation disrupts microtubule dynamics and podocyte functions in nonciliated cells, while in ciliated fibroblasts it leads to loss of cilia and impaired sonic hedgehog pathway. The NSL complex is a master regulator of intraciliary transport genes in both dividing and nondividing cells.\",\n      \"method\": \"Kidney-specific conditional knockout, single-cell RNA sequencing, cilia immunofluorescence, microtubule dynamics assay\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional KO in vivo with multiple defined cellular phenotypes and mechanistic pathway analysis (IFT, Shh, microtubule dynamics)\",\n      \"pmids\": [\"37624894\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Homozygous knockout of Kansl3 in mice leads to embryonic lethality at peri-implantation stages; embryos fail to hatch from the zona pellucida. Kansl3-null embryos have a significantly reduced inner cell mass (ICM) cell number with normal trophectoderm cell numbers, and both epiblast and primitive endoderm lineages show reduced cell numbers.\",\n      \"method\": \"Conditional/homozygous KO mouse, zona pellucida removal in vitro, immunofluorescence lineage marker analysis\",\n      \"journal\": \"Molecular reproduction and development\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO with defined lineage-specific cellular phenotype, single study\",\n      \"pmids\": [\"38769918\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"TRAP1 (molecular chaperone) directly interacts with KANSL3, as demonstrated by co-immunoprecipitation and LC-MS/MS. Under diabetic/high glucose-palmitate conditions, TRAP1-KANSL3 interaction decreases, and KANSL3 acetylation increases. TRAP1 inhibits KANSL3 acetylation under normal conditions to preserve mitophagy; loss of TRAP1 under diabetic conditions impairs mitophagy and mitochondrial function.\",\n      \"method\": \"Co-immunoprecipitation, LC-MS/MS, lentiviral TRAP1 overexpression and KANSL3 knockdown, mitophagy flux assay (mKeima), transmission electron microscopy\",\n      \"journal\": \"Cell communication and signaling : CCS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP with MS confirmation of interaction, functional rescue experiments; single lab study\",\n      \"pmids\": [\"41039555\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Hepatocyte-specific deletion of KANSL3 in mice results in early-onset liver disease marked by biliary hyperplasia and hepatic fibrosis. KANSL3 regulates hepatocyte transcriptional networks for hepatic steroid and lipid metabolism through histone acetylation. Loss of KANSL3 disrupts hepatocyte differentiation in vivo and impairs transcriptional programs for hepatocyte differentiation in ductal and fetal liver organoids.\",\n      \"method\": \"Hepatocyte-specific conditional KO, single-cell RNA sequencing, organoid differentiation assay, histone acetylation analysis\",\n      \"journal\": \"Life science alliance\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional KO in vivo with scRNA-seq, organoid functional rescue assays, and histone acetylation readouts across multiple orthogonal methods\",\n      \"pmids\": [\"41044006\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"KANSL3 is a core subunit of the evolutionarily conserved NSL histone acetyltransferase complex that associates with MOF/KAT8; during interphase it binds to gene promoters in a sequence-dependent manner to recruit the NURF remodeling complex, maintain nucleosome-depleted regions, and activate transcription of thousands of housekeeping and tissue-specific genes via H4K5/K8/K16 acetylation, while during mitosis it relocalizes to the spindle as a microtubule minus-end-binding and stabilizing factor essential for chromosome segregation; its protein stability is regulated by OGT1-mediated O-GlcNAcylation at Thr755 (opposing UBE2S-mediated ubiquitin degradation) and by TRAP1-mediated inhibition of its acetylation; loss of KANSL3 in vivo causes tissue-specific phenotypes including peri-implantation lethality, neurovascular breakdown through LCFA-TLR4-NFκB signaling, kidney dysfunction with impaired intraciliary transport, and liver disease through defective hepatocyte differentiation and lipid metabolism.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"KANSL3 is a core subunit of the evolutionarily conserved NSL histone acetyltransferase complex that associates with the acetyltransferase MOF and other NSL members to activate transcription genome-wide [#0]. At target promoters—predominantly TATA-less, housekeeping-gene promoters bound in a sequence-dependent manner—the complex maintains nucleosome-depleted regions and ensures accurate transcription start-site selection by recruiting the NURF chromatin remodeler, and is required for efficient assembly of the RNA Polymerase II pre-initiation complex (TBP, TFIIB) coincident with active chromatin marks including H4K16ac, H3K4me2/3, and H3K9ac [#1, #7]. KANSL3 carries out a moonlighting structural role outside chromatin: during mitosis it relocalizes to the spindle in a RanGTP-dependent manner, where it binds and stabilizes microtubule minus ends to enable spindle assembly and chromosome segregation [#2]. KANSL3 also partitions to mitochondria, where it is required for MOF binding to mtDNA and the consequent expression of respiratory genes [#3]. Its abundance and activity are post-translationally controlled: OGT1-mediated O-GlcNAcylation at Thr755 stabilizes KANSL3 and sustains NSL complex integrity and H4K5/K8/K16 acetylation, opposing UBE2S-driven ubiquitin-proteasomal degradation, with OGT1 and UBE2S competing for binding [#4, #5], while TRAP1 binding restrains KANSL3 acetylation to preserve mitophagy under metabolic stress [#14]. Through these transcriptional and structural roles, KANSL3 loss in vivo produces tissue-specific pathology—peri-implantation lethality with reduced inner cell mass [#13], neurovascular breakdown via LCFA-driven TLR4-NF\\u03baB signaling [#8], kidney dysfunction through disrupted intraciliary-transport gene programs and microtubule dynamics [#12], and liver disease through defective hepatocyte differentiation and lipid metabolism [#15].\",\n  \"teleology\": [\n    {\n      \"year\": 2010,\n      \"claim\": \"Established that KANSL3 is a bona fide subunit of the MOF-associated NSL complex and a positive regulator of genome-wide transcription, answering whether KANSL3 functions in chromatin-based gene activation.\",\n      \"evidence\": \"Biochemical purification, ChIP-Seq, RNAi depletion, and heterologous promoter tethering in Drosophila and mammals\",\n      \"pmids\": [\"20620954\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not define which promoter features dictate NSL targeting\", \"Direct enzymatic contribution of KANSL3 to acetylation not isolated from MOF\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Showed the NSL complex acts at the pre-initiation step by promoting RNA Pol II, TBP, and TFIIB recruitment at housekeeping-gene promoters, defining the transcriptional mechanism.\",\n      \"evidence\": \"ChIP-seq of NSL subunits and Pol II in NSL3-depleted Drosophila cells with RNAi knockdown\",\n      \"pmids\": [\"22723752\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of PIC stabilization not resolved\", \"Did not address non-chromatin roles\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Revealed a non-transcriptional structural function: KANSL3 relocalizes to the mitotic spindle and acts as a microtubule minus-end-binding stabilizer essential for chromosome segregation.\",\n      \"evidence\": \"Live-cell imaging, immunofluorescence, RanGTP-dependent spindle assembly assay, and loss-of-function mitotic readouts\",\n      \"pmids\": [\"26243146\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of minus-end binding not defined\", \"Relationship between chromatin and spindle pools unclear\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Demonstrated a mitochondrial pool of KANSL3 required for MOF–mtDNA association and respiratory gene expression, extending NSL function to oxidative phosphorylation.\",\n      \"evidence\": \"Subcellular fractionation, mtDNA ChIP, and conditional cardiac knockout mouse\",\n      \"pmids\": [\"27768893\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct mtDNA-binding activity of KANSL3 itself not shown\", \"How nuclear vs mitochondrial partitioning is controlled is unknown\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identified OGT1-mediated O-GlcNAcylation as a stabilizing modification of KANSL3 that boosts NSL acetyltransferase output on H4K5/K8/K16.\",\n      \"evidence\": \"Co-IP, in vitro O-GlcNAc transferase assay, WGA affinity purification, and OGT1 catalytic mutant in cells\",\n      \"pmids\": [\"28450392\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Modified residue not yet mapped at this stage\", \"In vivo physiological trigger of O-GlcNAcylation unknown\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Mapped the stabilizing O-GlcNAc site to Thr755 and showed OGT1 and UBE2S compete to set KANSL3 stability and NSL holoenzyme integrity, defining the degradation control axis.\",\n      \"evidence\": \"In vitro transferase assay with mass spectrometry, T755A mutagenesis, Co-IP, and ubiquitination assay\",\n      \"pmids\": [\"31881804\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"UBE2S substrate-recognition determinants not defined\", \"Upstream signals shifting the OGT1/UBE2S balance unknown\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Defined how NSL achieves nucleosome-free promoters and accurate TSS selection: KANSL3 binds TATA-less promoters sequence-dependently and recruits NURF to maintain nucleosome-depleted regions.\",\n      \"evidence\": \"ChIP-seq, nucleosome positioning after NSL1 depletion, and biochemical NSL–NURF interaction\",\n      \"pmids\": [\"30819819\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Sequence motif recognized by KANSL3 not specified\", \"Domain mediating NURF contact not mapped\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Indicated that mitotic defects after KANSL3-ortholog loss in Drosophila arise largely from transcriptional downregulation of centromere/kinetochore and centriole genes, linking transcriptional and mitotic phenotypes.\",\n      \"evidence\": \"RNAi depletion of Rcd1, immunofluorescence, GFP localization, and RT-qPCR in S2 cells\",\n      \"pmids\": [\"31527906\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single study; transcriptional vs direct spindle contributions not fully separated\", \"Centrosome/midbody localization function not mechanistically dissected\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Connected KANSL3-dependent epigenetic regulation to organ physiology by showing neural loss causes LCFA accumulation that drives TLR4-NF\\u03baB pericyte activation and neurovascular breakdown.\",\n      \"evidence\": \"Neural-specific conditional KO mice, metabolomics, TLR4 inhibitor rescue, and pericyte marker immunofluorescence\",\n      \"pmids\": [\"32541879\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct transcriptional targets governing LCFA metabolism not enumerated\", \"Cell-autonomous vs paracrine steps only partly separated\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identified YY1 as a key downstream transcriptional target in human cells, with KANSL3-driven clonogenicity rescued by YY1, placing YY1 epistatically below KANSL3.\",\n      \"evidence\": \"CRISPR/Cas9 KO, ChIP-seq, siRNA, overexpression, and colony formation in 293T/HepG2 cells\",\n      \"pmids\": [\"35409160\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab study\", \"Generalizability of YY1 axis to other cell types untested\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Showed KANSL3 loss promotes mesenchymal/invasive phenotypes, implicating NSL transcriptional regulation in epithelial cell identity.\",\n      \"evidence\": \"CRISPR/Cas9 KO, Transwell invasion assay, immunostaining, and ChIP-seq\",\n      \"pmids\": [\"35416545\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect control of mesenchymal markers unresolved\", \"Phosphoinositide-signaling link only correlative\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended NSL transcriptional function to small-RNA biology, showing KANSL3 promotes transcription of piRNA precursors and restrains transposons in the Drosophila germline.\",\n      \"evidence\": \"Germline-specific RNAi, small RNA sequencing, and ChIP-seq\",\n      \"pmids\": [\"37399316\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single study with effect described alongside other NSL subunits\", \"Conservation of piRNA role in mammals untested\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Established the NSL complex as a master regulator of intraciliary-transport genes and microtubule dynamics in both dividing and postmitotic cells, explaining kidney pathology on KANSL3 loss.\",\n      \"evidence\": \"Kidney-specific conditional KO, single-cell RNA-seq, cilia immunofluorescence, and microtubule dynamics assays\",\n      \"pmids\": [\"37624894\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether ciliary defect is purely transcriptional or also reflects direct microtubule role not fully separated\", \"Specific IFT target genes driving phenotype not pinpointed\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defined the earliest developmental requirement, showing Kansl3 is essential at peri-implantation with selective reduction of inner cell mass lineages.\",\n      \"evidence\": \"Homozygous KO mice, in vitro zona pellucida removal, and lineage-marker immunofluorescence\",\n      \"pmids\": [\"38769918\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular targets underlying ICM-specific defect not identified\", \"Single study\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identified TRAP1 as a direct KANSL3 partner that restrains its acetylation to preserve mitophagy, adding an acetylation-based control layer relevant to metabolic stress.\",\n      \"evidence\": \"Co-IP, LC-MS/MS, TRAP1 overexpression and KANSL3 knockdown, mKeima mitophagy flux, and TEM\",\n      \"pmids\": [\"41039555\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Acetyltransferase responsible for KANSL3 acetylation not identified\", \"Single-lab study; in vivo relevance untested\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Demonstrated that KANSL3 governs hepatocyte differentiation and lipid/steroid metabolic transcription, with loss causing biliary hyperplasia and fibrosis.\",\n      \"evidence\": \"Hepatocyte-specific conditional KO, single-cell RNA-seq, organoid differentiation, and histone acetylation analysis\",\n      \"pmids\": [\"41044006\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Key direct target genes of hepatocyte program not isolated\", \"Whether metabolic phenotype is cell-autonomous to hepatocytes not fully resolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How KANSL3 dynamically partitions among nuclear chromatin, mitotic spindle, and mitochondria, and what governs its tissue-specific target gene selection, remain open.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of KANSL3 domains for DNA, microtubule, and partner binding\", \"Mechanism of context-dependent localization switching unknown\", \"Sequence determinants of promoter targeting unmapped\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [7]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [0, 1, 7]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 1, 7]},\n      {\"term_id\": \"GO:0005819\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [0, 1, 7]},\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [7]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"complexes\": [\n      \"NSL (KANSL/MOF) histone acetyltransferase complex\"\n    ],\n    \"partners\": [\n      \"KAT8\",\n      \"OGT1\",\n      \"UBE2S\",\n      \"TRAP1\",\n      \"NURF\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}