{"gene":"MLLT10","run_date":"2026-06-10T02:59:50","timeline":{"discoveries":[{"year":1995,"finding":"AF10 protein contains a conserved cysteine-rich LAP/PHD zinc finger motif (now called LAP finger) that is proposed to be a DNA-binding domain; the LAP domain consensus sequence was defined by homology across 25+ proteins and the domain is disrupted in leukemia-associated chromosomal translocations.","method":"Sequence homology analysis and structural characterization of zinc finger motif","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Low","confidence_rationale":"Tier 4 / Weak — computational/sequence-based prediction only; no direct binding experiment performed on AF10 specifically","pmids":["7568208"],"is_preprint":false},{"year":1995,"finding":"In the t(10;11)(p12;q23) translocation causing AML, the leucine zipper motif of AF10 is consistently fused onto the N-terminal region of MLL/HRX, establishing that the leucine zipper of AF10 is a critical functional element juxtaposed onto MLL in all examined cases.","method":"Southern analysis and RT-PCR sequencing of leukemia patient samples","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple patient samples with consistent molecular breakpoint analysis, single lab","pmids":["7662954"],"is_preprint":false},{"year":2000,"finding":"AF10 contains an extended LAP/PHD finger domain that mediates homo-oligomerization of recombinant AF10 protein; AF10 also binds cruciform DNA via an AT-hook motif and is localized to the nucleus by a bipartite nuclear localization signal in its N-terminal region.","method":"Biochemical analysis of recombinant AF10 protein; GST pulldown for oligomerization; DNA-binding assay; subcellular fractionation","journal":"Journal of molecular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct biochemical reconstitution with recombinant protein, multiple orthogonal methods, single lab","pmids":["10860745"],"is_preprint":false},{"year":2001,"finding":"AF10 interacts with the Drosophila heterochromatin protein HP1 (in vitro and in vivo), and the Drosophila AF10 homolog dAF10 functions in heterochromatin-dependent gene silencing (position effect variegation), placing it in the HP1-dependent silencing pathway.","method":"In vitro binding assay and co-immunoprecipitation (in vivo); genetic analysis of position effect variegation in Drosophila","journal":"EMBO reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal in vitro and in vivo binding plus genetic functional assay, single lab, ortholog","pmids":["11266362"],"is_preprint":false},{"year":2002,"finding":"The AF10 leucine zipper domain interacts with GAS41 (a glioblastoma amplified gene product homologous to yeast ANC1); GAS41 in turn co-immunoprecipitates with INI1 (SNF5/SWI-SNF component), and INI1 is present in AF10 immunoprecipitates, placing AF10 in proximity to the SWI/SNF chromatin remodeling complex.","method":"Yeast two-hybrid screen; co-immunoprecipitation (in vivo confirmation)","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — yeast 2-hybrid plus reciprocal co-IP confirmation, single lab","pmids":["11756182"],"is_preprint":false},{"year":2002,"finding":"The AF10 leucine zipper (comprising two adjacent alpha-helical domains, 82 aa) is necessary and sufficient for MLL-AF10-mediated myeloid immortalization and leukemic transformation; deletion of the 29-aa leucine zipper within this region completely abrogates transforming activity; the same minimal domain also confers transcriptional activation when fused to MLL or GAL4 DNA-binding domains.","method":"Structure-function analysis with retroviral transduction of deletion mutants into primary murine myeloid progenitors; serial replating assays; in vivo leukemia model; GAL4 transcriptional reporter assay","journal":"Blood","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — multiple deletion mutants tested, in vitro and in vivo transformation assays, transcriptional readout, single lab with multiple orthogonal methods","pmids":["11986236"],"is_preprint":false},{"year":2001,"finding":"AF10 physically interacts with the synovial sarcoma-associated protein SYT; the interaction was mapped to the N-terminal region of SYT and a C-terminal region of AF10 outside known functional domains; co-localization confirmed in transfected cells.","method":"Yeast two-hybrid screen; co-immunoprecipitation of endogenous and epitope-tagged proteins; co-localization by immunofluorescence; sequential deletion mapping","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP plus localization and domain mapping, single lab","pmids":["11423977"],"is_preprint":false},{"year":2003,"finding":"The Drosophila AF10 homolog Alhambra (ALH) full-length protein has no activity on Polycomb group-responsive elements (PREs), but overexpression of the isolated leucine zipper domain activates several PREs; the PHD domain within the full-length protein inhibits the PRE-deregulating activity of the leucine zipper; this PRE deregulation is conserved in the human AF10 leucine zipper domain expressed in Drosophila.","method":"Drosophila genetics; PRE reporter assays; domain deletion/overexpression analysis in flies","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with domain-specific alleles in Drosophila ortholog, single lab","pmids":["12482966"],"is_preprint":false},{"year":2006,"finding":"CALM-AF10 fusion protein interacts with the H3K79 methyltransferase hDOT1L through the AF10 moiety; hDOT1L prevents nuclear export of CALM-AF10 and upregulates Hoxa5 through H3K79 methylation, contributing to leukemic transformation.","method":"Co-immunoprecipitation; ChIP for H3K79 methylation at Hoxa5 locus; retroviral bone marrow transformation assay; nuclear export assay","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP, ChIP showing H3K79me at specific locus, functional transformation assay, multiple orthogonal methods, single lab","pmids":["16921363"],"is_preprint":false},{"year":2006,"finding":"The CALM-AF10 fusion protein alters subcellular localization of the lymphoid transcription factor Ikaros by coexpression; the AF10 leucine zipper domain is required for the AF10-Ikaros interaction; the transcriptional repressor activity of Ikaros is reduced by AF10.","method":"Yeast two-hybrid screen; GST pulldown; co-immunoprecipitation; immunofluorescence co-localization; transcriptional reporter assay","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — GST pulldown plus co-IP plus localization plus reporter assay, single lab","pmids":["18037964"],"is_preprint":false},{"year":2006,"finding":"A novel CALM-interacting protein, CATS, increases the nuclear (and specifically nucleolar) localization of both CALM and the leukemogenic CALM/AF10 fusion protein; the CATS interaction domain was mapped to aa 221-335 of CALM.","method":"Yeast two-hybrid screen; GST pulldown; co-immunoprecipitation; co-localization by fluorescence microscopy; domain mapping","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal binding methods plus functional localization effect, single lab","pmids":["16491119"],"is_preprint":false},{"year":2009,"finding":"The CALM-AF10 fusion protein disrupts the normal AF10-mediated association of hDOT1L with chromatin, causing a global reduction of H3K79 methylation; cells with reduced H3K79 methylation show increased sensitivity to gamma-irradiation and chromosomal instability.","method":"ChIP analysis of H3K79 methylation; co-immunoprecipitation; gamma-irradiation sensitivity assays; cytogenetic analysis of leukemia patient samples","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP plus co-IP plus functional cellular assay, single lab","pmids":["19443658"],"is_preprint":false},{"year":2010,"finding":"MLLT10/AF10 and DOT1L are TCF4/β-catenin interactors in intestinal crypts; the AF10-DOT1L complex is recruited to Wnt target genes in a β-catenin-dependent manner and deposits H3K79 methylation over their coding regions; depletion of MLLT10/AF10 selectively impairs Wnt target gene expression and intestinal proliferation.","method":"Proteomics/mass spectrometry of TCF4/β-catenin complex; ChIP-H3K79me on Wnt target genes; siRNA knockdown with expression array; zebrafish morpholino experiments; genetic epistasis with apc-mutant zebrafish","journal":"PLoS biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — proteomics identification + ChIP + genetic epistasis + multiple model organisms + expression arrays, replicated across systems","pmids":["21103407"],"is_preprint":false},{"year":2011,"finding":"For CALM-AF10 leukemia transformation, the clathrin-binding domain of CALM (C-terminal 248 aa) and the octapeptide motif-leucine zipper (OM-LZ) domain of AF10 are the minimal sufficient domains; this 'minimal fusion' retains aberrant Hoxa cluster upregulation characteristic of full-length CALM-AF10.","method":"Structure-function analysis with CALM and AF10 domain deletion mutants; colony-forming assays; in vivo mouse leukemia model; gene expression analysis","journal":"Leukemia","confidence":"High","confidence_rationale":"Tier 2 / Strong — systematic domain deletion with in vitro and in vivo transformation assays, molecular phenotype confirmed, single lab","pmids":["21681188"],"is_preprint":false},{"year":2011,"finding":"Full-length CALM-AF10 localizes to the nucleus (not cytoplasm) in leukemia cells and has a propensity to homo-oligomerize as shown by FRET analysis; CALM-AF10 suppresses H3K79 methylation regardless of the presence of clathrin.","method":"Fluorescence microscopy; FRET analysis; ChIP for H3K79 methylation; mouse leukemia model with domain deletion constructs","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — FRET for oligomerization, ChIP for epigenetic mark, localization by imaging, single lab","pmids":["21706055"],"is_preprint":false},{"year":2012,"finding":"MLL-AF10 and CALM-AF10 mediated transformation is dependent on the H3K79 methyltransferase DOT1L; conditional Dot1l knockout abolishes in vitro transformation and in vivo leukemia initiation/maintenance; pharmacological DOT1L inhibition (EPZ004777) suppresses Hoxa cluster genes and Meis1, and selectively impairs proliferation of MLL-AF10 and CALM-AF10 transformed cells.","method":"Conditional Dot1l knockout mouse model; in vitro colony transformation assay; in vivo leukemia model; DOT1L inhibitor (EPZ004777) treatment; gene expression analysis","journal":"Leukemia","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — genetic knockout combined with pharmacological inhibition, in vitro and in vivo models, replicated across two fusion oncoproteins","pmids":["23138183"],"is_preprint":false},{"year":2012,"finding":"The PHD1-PHD2 module of the C. elegans AF10 ortholog ZFP-1 is essential for viability; the first PHD finger contributes to preferential binding of the PHD1-PHD2 domain to H3K4-methylated histone H3 tails; ZFP-1 occupancy genome-wide peaks at H3K4me-enriched promoters of active genes, and H3K4 methylation is required for ZFP-1 localization to promoters in the embryo.","method":"Genetic analysis (C. elegans deletion mutants); biochemical histone-tail binding assays; ChIP-seq for ZFP-1 and H3K4me marks","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — genetic viability assay, biochemical histone binding, genome-wide ChIP-seq, multiple orthogonal methods in the ortholog","pmids":["23263989"],"is_preprint":false},{"year":2013,"finding":"The C. elegans AF10 homolog ZFP-1 and its interacting partner DOT-1.1 globally reduce RNA Pol II transcription on essential widely expressed genes; the ZFP-1/DOT-1.1 complex increases H3K79 methylation and decreases H2B monoubiquitination (which promotes transcription), thereby negatively modulating Pol II elongation.","method":"Genomic (ChIP-seq, RNA-seq) and biochemical approaches; genetic knockdown; Pol II pausing analysis during development and stress response","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — genome-wide ChIP-seq, RNA-seq, biochemical co-complex analysis, multiple orthogonal methods in C. elegans ortholog","pmids":["23806335"],"is_preprint":false},{"year":2013,"finding":"CALM contains a CRM1-dependent nuclear export signal (NES) that mediates cytoplasmic localization of CALM-AF10 and is necessary for CALM-AF10-dependent transformation; NES motifs from heterologous proteins fused in-frame with AF10 are sufficient to immortalize murine hematopoietic progenitors; the CALM NES is essential for Hoxa gene upregulation and aberrant H3K79 methylation by CALM-AF10, possibly through mislocalization of DOT1L.","method":"NES mutational analysis; retroviral transformation assay; ChIP for H3K79me and HOXA gene expression; Leptomycin B (CRM1 inhibitor) treatment; heterologous NES fusion constructs","journal":"Blood","confidence":"High","confidence_rationale":"Tier 2 / Strong — systematic mutagenesis, functional transformation assay, pharmacological inhibition, ChIP validation, multiple orthogonal methods","pmids":["23487024"],"is_preprint":false},{"year":2014,"finding":"CRM1 physically localizes to HOXA loci and recruits CALM-AF10 to HOXA chromatin through the CALM NES-CRM1 interaction; genetic and pharmacological inhibition of CALM-CRM1 interaction prevents CALM-AF10 enrichment at HOXA chromatin and immediately abolishes HOXA transcription; CRM1 thus has a novel chromatin-binding and transcription factor-recruiting function.","method":"ChIP for CRM1 and CALM-AF10 at HOXA loci; genetic CALM NES mutants; CRM1 inhibitor treatment; transcriptional analysis","journal":"Leukemia","confidence":"High","confidence_rationale":"Tier 2 / Strong — ChIP showing CRM1 and CALM-AF10 co-occupancy, genetic NES mutants, pharmacological inhibition, multiple orthogonal methods","pmids":["25027513"],"is_preprint":false},{"year":2015,"finding":"The AF10 PZP domain (PHD finger-Zn knuckle-PHD finger module) reads unmodified H3K27; structural studies reveal that H3 binding triggers rearrangement of the PZP module to form an H3(22-27)-accommodating channel where unmodified H3K27 is encaged in a hydrogen-bond acceptor-lined cavity; H3K27 modification abrogates this interaction. In cells, PZP recognition of H3 is required for H3K79 dimethylation, expression of DOT1L-target genes, and proliferation of DOT1L-addicted leukemic cells.","method":"Crystal structure of PZP-H3 complex; mutagenesis of binding interface; histone peptide pulldowns; ChIP for H3K79me2 in cells with PZP mutants; cell proliferation assays","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure plus mutagenesis plus ChIP in cells plus functional proliferation assay, multiple orthogonal methods in single rigorous study","pmids":["26439302"],"is_preprint":false},{"year":2017,"finding":"Mllt10 knockout mice exhibit midline facial cleft associated with reduced H3K79 methylation in nasal processes; AP2α expression is directly regulated by Af10-dependent H3K79me at its locus, and is specifically reduced in nasal processes of Mllt10-KO embryos; pharmacological suppression of H3K79me completely phenocopies Mllt10-KO.","method":"Mllt10 knockout mouse; ChIP for H3K79me at AP2α locus in nasal processes; DOT1L inhibitor treatment phenocopy experiment; gene expression analysis","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO with defined molecular phenotype, ChIP at specific locus, pharmacological phenocopy, multiple orthogonal approaches","pmids":["28931923"],"is_preprint":false},{"year":2018,"finding":"Crystal structures of both apo AF10 OM-LZ and its complex with the coiled-coil domain of DOT1L were solved; disruption of the DOT1L-AF10 interface abrogates MLL-AF10-associated leukemic transformation; zinc stabilizes the DOT1L-AF10 complex and may regulate HOXA gene expression.","method":"X-ray crystallography of AF10 OM-LZ alone and in complex with DOT1L coiled-coil domain; interface mutagenesis; leukemic transformation assay","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure plus mutagenesis of interface plus functional leukemia transformation assay in single study","pmids":["29563185"],"is_preprint":false},{"year":2021,"finding":"AF10 fusions (PICALM-AF10 and MLL-AF10) directly recruit JAK1 kinase and activate a JAK/STAT-mediated inflammatory signaling cascade; genetic Jak1 deletion or pharmacological JAK/STAT inhibition elicits potent anti-oncogenic effects in mouse and human AF10-fusion AML models.","method":"Inducible mouse AML models; proteomics/protein interactome analysis; genetic Jak1 conditional knockout; pharmacological JAK/STAT inhibition; transcriptomic and epigenomic profiling","journal":"Blood","confidence":"High","confidence_rationale":"Tier 2 / Strong — proteomic identification of JAK1 as direct interactor, genetic KO, pharmacological inhibition, multiple AF10 fusions tested, multiple orthogonal approaches","pmids":["33690798"],"is_preprint":false},{"year":2021,"finding":"The AF10 PZP domain (AF10PZP) binds the nucleosome core particle through multivalent contacts with the histone H3 tail and DNA; AF10PZP associates with active chromatin marks and discriminates against H3K27me3; disruption of AF10PZP function in CALM-AF10 drives transformation, while incorporation of functional AF10PZP into CALM-AF10 prevents transformation, promotes nuclear localization, and downregulates Hoxa genes.","method":"Crystallography of AF10PZP-nucleosome complex; biochemical binding assays; mutagenesis; bone marrow transformation assays in vitro and in vivo; ChIP; gene expression analysis","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure, biochemical assays, mutagenesis, in vitro and in vivo functional transformation assays, ChIP, multiple orthogonal methods","pmids":["34226546"],"is_preprint":false},{"year":2021,"finding":"AF10 (MLLT10) is required for higher-order H3K79 methylation (H3K79me2/me3) in somatic cells; suppression of AF10 via RNAi or CRISPR/Cas9 significantly increases somatic cell reprogramming efficiency; re-expression of wild-type AF10 but not a DOT1L-binding-impaired AF10 mutant rescues H3K79 methylation and reduces reprogramming efficiency, establishing AF10 as a barrier to reprogramming through its regulation of DOT1L-dependent H3K79 methylation.","method":"Proximity-based labeling proteomics (BioID); RNAi and CRISPR/Cas9 knockdown; reprogramming efficiency assay; rescue with wild-type vs. DOT1L-binding mutant AF10; H3K79me western blot; transcriptomic analysis","journal":"Epigenetics & chromatin","confidence":"High","confidence_rationale":"Tier 2 / Strong — proteomics identification, genetic KO, structure-function rescue with DOT1L-binding mutant, multiple orthogonal approaches in single study","pmids":["34215314"],"is_preprint":false},{"year":2021,"finding":"Tip60 histone acetyltransferase is recruited by MLL-AF10 to the Hoxa9 locus where it acetylates H2A.Z to promote Hoxa9 gene expression; conditional deletion of Tip60 prevents development of MLL-AF10 leukemia.","method":"ChIP showing Tip60 recruitment to Hoxa9 by MLL-AF10; H2A.Z acetylation assay; conditional Tip60 knockout; leukemia development assay in mice","journal":"Leukemia","confidence":"High","confidence_rationale":"Tier 2 / Strong — ChIP, histone modification assay, genetic conditional KO with functional leukemia phenotype, multiple orthogonal methods","pmids":["33967269"],"is_preprint":false},{"year":2023,"finding":"DOT1L associates with MLLT10 in testis; DOT1L and MLLT10 co-localize to sex chromatin in meiotic and post-meiotic germ cells in an inter-dependent manner; both DOT1L and MLLT10 are essential for H3K79me2 in germ cells and for histone-to-protamine replacement during spermiogenesis; loss of either DOT1L or MLLT10 results in histone retention in sperm and male subfertility.","method":"Co-immunoprecipitation (DOT1L-MLLT10 in testis); immunofluorescence co-localization in germ cells; Mllt10 conditional knockout mouse; H3K79me2 ChIP/immunostaining; sperm histone retention assay","journal":"Development (Cambridge, England)","confidence":"High","confidence_rationale":"Tier 2 / Strong — co-IP, co-localization, conditional KO with defined epigenetic and developmental phenotype, multiple orthogonal methods","pmids":["37082953"],"is_preprint":false},{"year":2023,"finding":"AF10 deletion evicts H3K79me2/3 from gene bodies and redistributes H3K79me1 to transcription start sites; AF10 loss also redistributes RNA Pol II to a pattern characteristic of pluripotency at highly expressed housekeeping genes, facilitating iPSC formation without major steady-state transcriptional changes; this reveals that AF10 controls the patterning of H3K79 methylation orders to maintain cell identity.","method":"AF10 genetic deletion (CRISPR); CUT&RUN/ChIP for H3K79me1/me2/me3; RNA Pol II ChIP-seq; reprogramming efficiency assay; DOT1L chemical inhibition as comparison","journal":"Stem cell reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO, genome-wide ChIP-seq for multiple histone marks and Pol II, reprogramming assay, chemical inhibition orthogonal comparison","pmids":["37995701"],"is_preprint":false},{"year":2007,"finding":"The nuclear form of FLRG (follistatin-related gene) interacts with AF10 via the N-terminal PHD-containing region of AF10; FLRG enhances AF10 homo-oligomerization and augments the transcriptional activation properties of AF10 in reporter assays.","method":"Yeast two-hybrid screen; far-Western blot; co-immunoprecipitation; transcriptional reporter (Gal4-AF10 fusion); domain mapping","journal":"Biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple binding methods (Y2H, far-Western, co-IP) plus functional reporter, single lab","pmids":["17868029"],"is_preprint":false}],"current_model":"MLLT10/AF10 is a transcriptional co-regulator that functions primarily as a cofactor for the H3K79 methyltransferase DOT1L: AF10 binds DOT1L through its octapeptide motif-leucine zipper (OM-LZ) domain (structurally resolved by X-ray crystallography), while its N-terminal PZP domain (PHD-Zn knuckle-PHD) reads unmodified H3K27 on nucleosomes to target DOT1L-dependent H3K79 dimethylation and trimethylation to gene bodies of active genes; AF10 also homo-oligomerizes via its extended LAP/PHD finger, binds chromatin via an AT-hook, localizes to the nucleus via a bipartite NLS, and associates with the SWI/SNF complex through GAS41 and INI1; in development, AF10-dependent H3K79 methylation regulates AP2α expression in nasal processes and histone-to-protamine replacement during spermiogenesis; in Wnt signaling, AF10-DOT1L is recruited by TCF4/β-catenin to deposit H3K79me over Wnt target gene coding regions; in leukemia, AF10 fusion proteins (MLL-AF10, CALM-AF10) mislocalize or misdirect DOT1L to HOXA loci, driving aberrant H3K79 methylation and HOXA cluster gene upregulation, and also directly recruit JAK1 to activate JAK/STAT inflammatory signaling."},"narrative":{"mechanistic_narrative":"MLLT10/AF10 is a nuclear chromatin-associated cofactor that targets the H3K79 methyltransferase DOT1L to active chromatin, governing transcriptional programs in development, stem cell identity, and leukemia [PMID:23138183, PMID:26439302, PMID:37995701]. AF10 binds DOT1L directly through its octapeptide motif-leucine zipper (OM-LZ) domain, an interaction resolved by crystallography and stabilized by zinc, and disruption of this interface abolishes MLL-AF10 leukemic transformation [PMID:29563185]. Its N-terminal PZP module (PHD-Zn knuckle-PHD) engages the nucleosome through multivalent contacts with unmodified H3 (specifically unmethylated H3K27) and DNA, discriminating against repressive H3K27me3, thereby directing DOT1L-dependent H3K79 di- and trimethylation to gene bodies of active genes; PZP recognition is required for H3K79me2 deposition, DOT1L-target gene expression, and proliferation of DOT1L-addicted cells [PMID:26439302, PMID:34226546]. AF10 controls the higher-order patterning of H3K79 methylation across the genome and the redistribution of RNA Pol II, acting as a barrier to somatic cell reprogramming that maintains cell identity [PMID:34215314, PMID:37995701]. Through this DOT1L axis, AF10 directly regulates AP2α expression and midline facial development [PMID:28931923] and drives histone-to-protamine replacement during spermiogenesis, where DOT1L and MLLT10 co-localize to sex chromatin and are mutually required for germ-cell H3K79me2 [PMID:37082953]. In Wnt signaling, the AF10-DOT1L complex is recruited by TCF4/β-catenin to deposit H3K79 methylation over Wnt target gene coding regions, supporting intestinal proliferation [PMID:21103407]. In leukemia, AF10 fusion oncoproteins (MLL-AF10, CALM/PICALM-AF10) misdirect DOT1L to drive aberrant H3K79 methylation and HOXA cluster upregulation; the minimal transforming unit is the AF10 OM-LZ together with the CALM clathrin-binding/NES region, with the CALM NES recruiting CRM1 to HOXA chromatin [PMID:21681188, PMID:23138183, PMID:23487024, PMID:25027513]. AF10 fusions additionally recruit Tip60 to acetylate H2A.Z at Hoxa9 and directly recruit JAK1 to activate JAK/STAT inflammatory signaling, both of which are required for leukemogenesis [PMID:33690798, PMID:33967269].","teleology":[{"year":1995,"claim":"Established AF10's domain architecture and its central role in leukemia, defining the LAP/PHD zinc finger and identifying the leucine zipper as the element consistently fused to MLL in t(10;11) AML.","evidence":"Sequence homology analysis of zinc finger motifs and breakpoint mapping in AML patient samples","pmids":["7568208","7662954"],"confidence":"Medium","gaps":["LAP/PHD DNA-binding was predicted, not demonstrated","no mechanistic link to a methyltransferase yet","functional consequence of the leucine zipper fusion unresolved"]},{"year":2000,"claim":"Defined the biochemical properties of AF10 domains, showing the extended LAP/PHD finger drives homo-oligomerization, an AT-hook binds cruciform DNA, and a bipartite NLS directs nuclear localization.","evidence":"Biochemical reconstitution with recombinant AF10, GST pulldown, DNA-binding assay, and subcellular fractionation","pmids":["10860745"],"confidence":"Medium","gaps":["physiological relevance of cruciform DNA binding unclear","oligomerization partners in cells not defined","no chromatin target identified"]},{"year":2001,"claim":"Linked AF10 to chromatin silencing and other binding partners, connecting it to HP1-dependent heterochromatin in Drosophila and identifying SYT and (later) FLRG as interactors.","evidence":"In vitro/in vivo binding, co-IP, position-effect-variegation genetics in Drosophila, and yeast two-hybrid with domain mapping","pmids":["11266362","11423977","17868029"],"confidence":"Medium","gaps":["mechanistic role of HP1, SYT, FLRG interactions in mammalian AF10 function unestablished","ortholog-based silencing data may not transfer directly","FLRG/SYT not connected to the DOT1L axis"]},{"year":2002,"claim":"Showed the AF10 leucine zipper is necessary and sufficient for MLL-AF10 transformation and confers transcriptional activation, and placed AF10 in proximity to SWI/SNF via GAS41 and INI1.","evidence":"Deletion-mutant retroviral transformation in murine progenitors, serial replating, GAL4 reporter assays, and yeast two-hybrid/co-IP","pmids":["11986236","11756182"],"confidence":"High","gaps":["the leucine-zipper effector partner was not yet identified","relationship between SWI/SNF association and transformation unclear"]},{"year":2006,"claim":"Identified DOT1L as the key AF10 effector in leukemia, showing CALM-AF10 binds hDOT1L through the AF10 moiety to drive H3K79 methylation and Hoxa upregulation.","evidence":"Co-IP, ChIP for H3K79me at Hoxa5, nuclear export assay, and retroviral transformation; plus Ikaros and CATS interaction mapping","pmids":["16921363","18037964","16491119"],"confidence":"High","gaps":["how AF10 normally targets DOT1L to chromatin not yet defined","global versus local H3K79me effects unresolved"]},{"year":2009,"claim":"Revealed that CALM-AF10 disrupts normal AF10-mediated DOT1L-chromatin association, causing global H3K79me loss, genomic instability, and irradiation sensitivity.","evidence":"ChIP for H3K79me, co-IP, gamma-irradiation sensitivity assays, and cytogenetics of patient samples","pmids":["19443658"],"confidence":"Medium","gaps":["reconciling global H3K79me loss with locus-specific HOXA gain unresolved","molecular basis of mistargeting not yet structural"]},{"year":2010,"claim":"Defined a physiological developmental role by showing the AF10-DOT1L complex is recruited by TCF4/β-catenin to deposit H3K79me over Wnt target gene coding regions and drive intestinal proliferation.","evidence":"TCF4/β-catenin proteomics, ChIP-H3K79me, siRNA expression arrays, and zebrafish epistasis with apc mutants","pmids":["21103407"],"confidence":"High","gaps":["direct contacts between AF10 and the TCF4/β-catenin module not mapped","selectivity for Wnt over other active genes unexplained"]},{"year":2011,"claim":"Defined the minimal CALM-AF10 transforming unit (CALM clathrin-binding domain plus AF10 OM-LZ) and showed full-length CALM-AF10 localizes to the nucleus, homo-oligomerizes, and suppresses H3K79me independent of clathrin.","evidence":"Domain-deletion colony and mouse leukemia assays, FRET oligomerization analysis, and ChIP for H3K79me","pmids":["21681188","21706055"],"confidence":"High","gaps":["mechanism connecting oligomerization to H3K79me dysregulation unclear","role of CALM moiety in DOT1L mistargeting not yet defined"]},{"year":2012,"claim":"Established DOT1L as the genetic and pharmacological dependency of AF10-fusion leukemia and characterized AF10/DOT1L as a repressive elongation modulator in the ortholog.","evidence":"Conditional Dot1l knockout, EPZ004777 inhibition, and in vitro/in vivo leukemia models; plus C. elegans ZFP-1/DOT-1.1 ChIP-seq/RNA-seq","pmids":["23138183","23806335","23263989"],"confidence":"High","gaps":["ortholog ZFP-1 repressive role contrasts with activating mammalian role, leaving directionality context-dependent","PHD-module H3K4me reading shown in ortholog, not yet human"]},{"year":2014,"claim":"Resolved how CALM-AF10 reaches HOXA chromatin, showing the CALM CRM1-dependent NES is required for transformation and recruits CRM1, which itself occupies HOXA loci and brings in CALM-AF10.","evidence":"NES mutagenesis, heterologous NES fusions, Leptomycin B treatment, and ChIP for CRM1 and CALM-AF10 at HOXA","pmids":["23487024","25027513"],"confidence":"High","gaps":["how CRM1 selects HOXA chromatin not defined","generalizability beyond CALM-AF10 to MLL-AF10 unclear"]},{"year":2015,"claim":"Provided the structural basis for AF10 chromatin targeting, showing the PZP module reads unmodified H3K27 to license DOT1L-dependent H3K79me2 and DOT1L-target gene expression.","evidence":"Crystal structure of PZP-H3 complex, interface mutagenesis, histone peptide pulldowns, and ChIP/proliferation assays in cells","pmids":["26439302"],"confidence":"High","gaps":["nucleosomal (versus peptide) engagement not yet resolved at this stage","interplay between PZP reading and OM-LZ DOT1L binding not integrated"]},{"year":2017,"claim":"Demonstrated a non-leukemic developmental requirement, with Mllt10 knockout causing midline facial cleft through loss of Af10-dependent H3K79me at the AP2α locus.","evidence":"Mllt10 knockout mouse, ChIP for H3K79me at AP2α in nasal processes, and DOT1L-inhibitor phenocopy","pmids":["28931923"],"confidence":"High","gaps":["other AF10-dependent developmental target genes not catalogued","tissue-specific targeting mechanism unexplained"]},{"year":2018,"claim":"Provided atomic detail of the AF10-DOT1L interface, solving the OM-LZ/DOT1L coiled-coil complex, showing zinc stabilization, and proving the interface is required for MLL-AF10 transformation.","evidence":"X-ray crystallography of AF10 OM-LZ alone and with DOT1L coiled-coil, interface mutagenesis, and leukemic transformation assay","pmids":["29563185"],"confidence":"High","gaps":["functional role of zinc regulation in normal cells not defined","whether the interface is druggable in leukemia untested here"]},{"year":2021,"claim":"Integrated nucleosome-level PZP recognition, oncogenic JAK/STAT and Tip60 axes, and a reprogramming-barrier role, establishing AF10 as a multivalent chromatin reader controlling H3K79me-dependent cell identity and additional leukemogenic signaling.","evidence":"PZP-nucleosome crystallography with transformation rescue, BioID proteomics with reprogramming assays, and proteomics/genetic-pharmacological dissection of JAK1 and Tip60 in AML models","pmids":["34226546","34215314","33690798","33967269"],"confidence":"High","gaps":["how JAK1 recruitment relates mechanistically to H3K79me deposition unclear","whether Tip60/H2A.Z acetylation acts upstream or in parallel to DOT1L unresolved"]},{"year":2023,"claim":"Showed AF10 governs the higher-order patterning of H3K79 methylation and Pol II distribution to maintain cell identity and, in testis, partners with DOT1L for histone-to-protamine replacement during spermiogenesis.","evidence":"AF10 CRISPR deletion with CUT&RUN/ChIP-seq for H3K79me1/2/3 and Pol II plus reprogramming assays; and testis co-IP, co-localization, and Mllt10 conditional knockout with sperm histone-retention readout","pmids":["37995701","37082953"],"confidence":"High","gaps":["how AF10 selects which H3K79 methylation order to deposit at a given locus unknown","mechanism coupling H3K79me patterning to Pol II redistribution undefined"]},{"year":null,"claim":"How AF10's two chromatin-engaging modules (PZP nucleosome reading and OM-LZ DOT1L binding) are coordinated to choose specific loci and to set the order of H3K79 methylation, and how the non-DOT1L oncogenic activities (JAK1, Tip60, CRM1) are mechanistically integrated, remains unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["no unified model linking PZP/OM-LZ engagement to locus selection","mechanism setting H3K79me1 vs me2/me3 at a locus unknown","integration of JAK/STAT, Tip60, and CRM1 axes with the DOT1L methyltransferase axis undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0042393","term_label":"histone binding","supporting_discovery_ids":[20,24,16]},{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[2,24]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[5,12,29]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[20,22,24]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[2,14]},{"term_id":"GO:0000228","term_label":"nuclear chromosome","supporting_discovery_ids":[20,24,28]}],"pathway":[{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[20,24,28]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[12,21]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[15,18,22,23]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[12,23]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[21,27]}],"complexes":["AF10-DOT1L complex","SWI/SNF (proximity via GAS41/INI1)"],"partners":["DOT1L","GAS41","INI1/SMARCB1","JAK1","TCF4","CTNNB1","KAT5/TIP60","CRM1/XPO1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P55197","full_name":"Protein AF-10","aliases":["ALL1-fused gene from chromosome 10 protein"],"length_aa":1068,"mass_kda":113.3,"function":"Probably involved in transcriptional regulation. In vitro or as fusion protein with KMT2A/MLL1 has transactivation activity. Binds to cruciform DNA. In cells, binding to unmodified histone H3 regulates DOT1L functions including histone H3 'Lys-79' dimethylation (H3K79me2) and gene activation (PubMed:26439302)","subcellular_location":"Nucleus","url":"https://www.uniprot.org/uniprotkb/P55197/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MLLT10","classification":"Not Classified","n_dependent_lines":7,"n_total_lines":1208,"dependency_fraction":0.005794701986754967},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"HIST2H2BE","stoichiometry":0.2},{"gene":"SSRP1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/MLLT10","total_profiled":1310},"omim":[{"mim_id":"612708","title":"RANBP2-LIKE AND GRIP DOMAIN-CONTAINING PROTEIN 5; RGPD5","url":"https://www.omim.org/entry/612708"},{"mim_id":"608757","title":"CLEAVAGE FACTOR POLYNUCLEOTIDE KINASE SUBUNIT 1; CLP1","url":"https://www.omim.org/entry/608757"},{"mim_id":"607375","title":"DOT1-LIKE; DOT1L","url":"https://www.omim.org/entry/607375"},{"mim_id":"607174","title":"MENINGIOMA, FAMILIAL, SUSCEPTIBILITY TO","url":"https://www.omim.org/entry/607174"},{"mim_id":"602409","title":"MLLT10 HISTONE LYSINE METHYLTRANSFERASE DOT1L COFACTOR; MLLT10","url":"https://www.omim.org/entry/602409"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"testis","ntpm":86.9}],"url":"https://www.proteinatlas.org/search/MLLT10"},"hgnc":{"alias_symbol":["AF10"],"prev_symbol":[]},"alphafold":{"accession":"P55197","domains":[{"cath_id":"3.30.40.10","chopping":"40-208","consensus_level":"medium","plddt":92.4282,"start":40,"end":208}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P55197","model_url":"https://alphafold.ebi.ac.uk/files/AF-P55197-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P55197-F1-predicted_aligned_error_v6.png","plddt_mean":50.03},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MLLT10","jax_strain_url":"https://www.jax.org/strain/search?query=MLLT10"},"sequence":{"accession":"P55197","fasta_url":"https://rest.uniprot.org/uniprotkb/P55197.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P55197/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P55197"}},"corpus_meta":[{"pmid":"8643484","id":"PMC_8643484","title":"The t(10;11)(p13;q14) in the U937 cell line results in the fusion of the AF10 gene and CALM, encoding a new member of the AP-3 clathrin assembly protein family.","date":"1996","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/8643484","citation_count":268,"is_preprint":false},{"pmid":"16921363","id":"PMC_16921363","title":"Leukaemic transformation by CALM-AF10 involves upregulation of Hoxa5 by hDOT1L.","date":"2006","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/16921363","citation_count":147,"is_preprint":false},{"pmid":"7568208","id":"PMC_7568208","title":"The leukemia-associated-protein (LAP) domain, a cysteine-rich motif, is present in a wide range of proteins, including MLL, AF10, and MLLT6 proteins.","date":"1995","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/7568208","citation_count":137,"is_preprint":false},{"pmid":"7662954","id":"PMC_7662954","title":"The t(10;11) translocation in acute myeloid leukemia (M5) consistently fuses the leucine zipper motif of AF10 onto the HRX gene.","date":"1995","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/7662954","citation_count":115,"is_preprint":false},{"pmid":"23138183","id":"PMC_23138183","title":"Abrogation of MLL-AF10 and CALM-AF10-mediated transformation through genetic inactivation or pharmacological inhibition of the H3K79 methyltransferase Dot1l.","date":"2012","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/23138183","citation_count":114,"is_preprint":false},{"pmid":"12676784","id":"PMC_12676784","title":"CALM-AF10 is a common fusion transcript in T-ALL and is specific to the TCRgammadelta lineage.","date":"2003","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/12676784","citation_count":114,"is_preprint":false},{"pmid":"11986236","id":"PMC_11986236","title":"The AF10 leucine zipper is required for leukemic transformation of myeloid progenitors by MLL-AF10.","date":"2002","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/11986236","citation_count":112,"is_preprint":false},{"pmid":"17097559","id":"PMC_17097559","title":"Acute myeloid leukemia is propagated by a leukemic stem cell with lymphoid characteristics in a mouse model of CALM/AF10-positive leukemia.","date":"2006","source":"Cancer cell","url":"https://pubmed.ncbi.nlm.nih.gov/17097559","citation_count":99,"is_preprint":false},{"pmid":"7671224","id":"PMC_7671224","title":"Breakpoint heterogeneity in t(10;11) translocation in AML-M4/M5 resulting in fusion of AF10 and MLL is resolved by fluorescent in situ hybridization analysis.","date":"1995","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/7671224","citation_count":79,"is_preprint":false},{"pmid":"11756182","id":"PMC_11756182","title":"The MLL fusion partner AF10 binds GAS41, a protein that interacts with the human SWI/SNF complex.","date":"2002","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/11756182","citation_count":76,"is_preprint":false},{"pmid":"21103407","id":"PMC_21103407","title":"The leukemia-associated Mllt10/Af10-Dot1l are Tcf4/β-catenin coactivators essential for intestinal homeostasis.","date":"2010","source":"PLoS biology","url":"https://pubmed.ncbi.nlm.nih.gov/21103407","citation_count":75,"is_preprint":false},{"pmid":"10637482","id":"PMC_10637482","title":"Molecular analysis of the CALM/AF10 fusion: identical rearrangements in acute myeloid leukemia, acute lymphoblastic leukemia and malignant lymphoma patients.","date":"2000","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/10637482","citation_count":72,"is_preprint":false},{"pmid":"8388418","id":"PMC_8388418","title":"Recombinant IL-6 activates p42 and p44 mitogen-activated protein kinases in the IL-6 responsive B cell line, AF-10.","date":"1993","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/8388418","citation_count":72,"is_preprint":false},{"pmid":"26439302","id":"PMC_26439302","title":"The PZP Domain of AF10 Senses Unmodified H3K27 to Regulate DOT1L-Mediated Methylation of H3K79.","date":"2015","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/26439302","citation_count":68,"is_preprint":false},{"pmid":"18094714","id":"PMC_18094714","title":"The role of CALM-AF10 gene fusion in acute leukemia.","date":"2007","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/18094714","citation_count":64,"is_preprint":false},{"pmid":"10860745","id":"PMC_10860745","title":"Biochemical analyses of the AF10 protein: the extended LAP/PHD-finger mediates oligomerisation.","date":"2000","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/10860745","citation_count":64,"is_preprint":false},{"pmid":"17804713","id":"PMC_17804713","title":"Expression of a CALM-AF10 fusion gene leads to Hoxa cluster overexpression and acute leukemia in transgenic mice.","date":"2007","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/17804713","citation_count":59,"is_preprint":false},{"pmid":"23673860","id":"PMC_23673860","title":"New MLLT10 gene recombinations in pediatric T-acute lymphoblastic leukemia.","date":"2013","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/23673860","citation_count":55,"is_preprint":false},{"pmid":"10554802","id":"PMC_10554802","title":"Consistent detection of CALM-AF10 chimaeric transcripts in haematological malignancies with t(10;11)(p13;q14) and identification of novel transcripts.","date":"1999","source":"British journal of haematology","url":"https://pubmed.ncbi.nlm.nih.gov/10554802","citation_count":51,"is_preprint":false},{"pmid":"16491119","id":"PMC_16491119","title":"The novel CALM interactor CATS influences the subcellular localization of the leukemogenic fusion protein CALM/AF10.","date":"2006","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/16491119","citation_count":50,"is_preprint":false},{"pmid":"19443658","id":"PMC_19443658","title":"Global reduction of the epigenetic H3K79 methylation mark and increased chromosomal instability in CALM-AF10-positive leukemias.","date":"2009","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/19443658","citation_count":50,"is_preprint":false},{"pmid":"23806335","id":"PMC_23806335","title":"The ZFP-1(AF10)/DOT-1 complex opposes H2B ubiquitination to reduce Pol II transcription.","date":"2013","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/23806335","citation_count":46,"is_preprint":false},{"pmid":"22871473","id":"PMC_22871473","title":"PICALM-MLLT10 acute myeloid leukemia: a French cohort of 18 patients.","date":"2012","source":"Leukemia research","url":"https://pubmed.ncbi.nlm.nih.gov/22871473","citation_count":45,"is_preprint":false},{"pmid":"10221337","id":"PMC_10221337","title":"Mixed-lineage leukemia with t(10;11)(p13;q21): an analysis of AF10-CALM and CALM-AF10 fusion mRNAs and clinical features.","date":"1999","source":"Genes, chromosomes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/10221337","citation_count":42,"is_preprint":false},{"pmid":"11423977","id":"PMC_11423977","title":"The synovial sarcoma associated protein SYT interacts with the acute leukemia associated protein AF10.","date":"2001","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/11423977","citation_count":41,"is_preprint":false},{"pmid":"25027513","id":"PMC_25027513","title":"A critical role for CRM1 in regulating HOXA gene transcription in CALM-AF10 leukemias.","date":"2014","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/25027513","citation_count":39,"is_preprint":false},{"pmid":"22064352","id":"PMC_22064352","title":"CALM/AF10-positive leukemias show upregulation of genes involved in chromatin assembly and DNA repair processes and of genes adjacent to the breakpoint at 10p12.","date":"2011","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/22064352","citation_count":38,"is_preprint":false},{"pmid":"19383357","id":"PMC_19383357","title":"The CALM and CALM/AF10 interactor CATS is a marker for proliferation.","date":"2008","source":"Molecular oncology","url":"https://pubmed.ncbi.nlm.nih.gov/19383357","citation_count":38,"is_preprint":false},{"pmid":"21681188","id":"PMC_21681188","title":"The clathrin-binding domain of CALM and the OM-LZ domain of AF10 are sufficient to induce acute myeloid leukemia in mice.","date":"2011","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/21681188","citation_count":33,"is_preprint":false},{"pmid":"18037964","id":"PMC_18037964","title":"The leukemogenic CALM/AF10 fusion protein alters the subcellular localization of the lymphoid regulator Ikaros.","date":"2007","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/18037964","citation_count":31,"is_preprint":false},{"pmid":"23487024","id":"PMC_23487024","title":"A CALM-derived nuclear export signal is essential for CALM-AF10-mediated leukemogenesis.","date":"2013","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/23487024","citation_count":31,"is_preprint":false},{"pmid":"8896421","id":"PMC_8896421","title":"AF10 is split by MLL and HEAB, a human homolog to a putative Caenorhabditis elegans ATP/GTP-binding protein in an invins(10;11)(p12;q23q12).","date":"1996","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/8896421","citation_count":30,"is_preprint":false},{"pmid":"30707474","id":"PMC_30707474","title":"Acute leukemias harboring KMT2A/MLLT10 fusion: a 10-year experience from a single genomics laboratory.","date":"2019","source":"Genes, chromosomes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/30707474","citation_count":27,"is_preprint":false},{"pmid":"23564351","id":"PMC_23564351","title":"Involvement of Gpr125 in the myeloid sarcoma formation induced by cooperating MLL/AF10(OM-LZ) and oncogenic KRAS in a mouse bone marrow transplantation model.","date":"2013","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/23564351","citation_count":27,"is_preprint":false},{"pmid":"12482966","id":"PMC_12482966","title":"The leucine zipper motif of the Drosophila AF10 homologue can inhibit PRE-mediated repression: implications for leukemogenic activity of human MLL-AF10 fusions.","date":"2003","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/12482966","citation_count":27,"is_preprint":false},{"pmid":"9737689","id":"PMC_9737689","title":"Alternative splicing in wild-type AF10 and CALM cDNAs and in AF10-CALM and CALM-AF10 fusion cDNAs produced by the t(10;11)(p13-14;q14-q21) suggests a potential role for truncated AF10 polypeptides.","date":"1998","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/9737689","citation_count":25,"is_preprint":false},{"pmid":"11477655","id":"PMC_11477655","title":"t(10;11)-acute leukemias with MLL-AF10 and MLL-ABI1 chimeric transcripts: specific expression patterns of ABI1 gene in leukemia and solid tumor cell lines.","date":"2001","source":"Genes, chromosomes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/11477655","citation_count":24,"is_preprint":false},{"pmid":"20875875","id":"PMC_20875875","title":"Acute leukemia with PICALM-MLLT10 fusion gene: diagnostic and treatment struggle.","date":"2010","source":"Cancer genetics and cytogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/20875875","citation_count":23,"is_preprint":false},{"pmid":"11266362","id":"PMC_11266362","title":"The Drosophila homolog of the human AF10 is an HP1-interacting suppressor of position effect variegation.","date":"2001","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/11266362","citation_count":23,"is_preprint":false},{"pmid":"15135402","id":"PMC_15135402","title":"Purification and characterization of inulinase from Aspergillus niger AF10 expressed in Pichia pastoris.","date":"2004","source":"Protein expression and purification","url":"https://pubmed.ncbi.nlm.nih.gov/15135402","citation_count":23,"is_preprint":false},{"pmid":"9878787","id":"PMC_9878787","title":"Expression pattern and cellular distribution of the murine homologue of AF10.","date":"1998","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/9878787","citation_count":22,"is_preprint":false},{"pmid":"29563185","id":"PMC_29563185","title":"Structural and functional analysis of the DOT1L-AF10 complex reveals mechanistic insights into MLL-AF10-associated leukemogenesis.","date":"2018","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/29563185","citation_count":21,"is_preprint":false},{"pmid":"32765078","id":"PMC_32765078","title":"miR-331-3p Inhibits Tumor Cell Proliferation, Metastasis, Invasion by Targeting MLLT10 in Non-Small Cell Lung Cancer.","date":"2020","source":"Cancer management and research","url":"https://pubmed.ncbi.nlm.nih.gov/32765078","citation_count":21,"is_preprint":false},{"pmid":"12810254","id":"PMC_12810254","title":"Cytogenetics, fluorescence in situ hybridization, and reverse transcriptase polymerase chain reaction are necessary to clarify the various mechanisms leading to an MLL-AF10 fusion in acute myelocytic leukemia with 10;11 rearrangement.","date":"2003","source":"Cancer genetics and cytogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/12810254","citation_count":20,"is_preprint":false},{"pmid":"10602503","id":"PMC_10602503","title":"The cloning, mapping and expression of a novel gene, BRL, related to the AF10 leukaemia gene.","date":"1999","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/10602503","citation_count":19,"is_preprint":false},{"pmid":"32033330","id":"PMC_32033330","title":"The Association of the Copy Number Variation of the MLLT10 Gene with Growth Traits of Chinese Cattle.","date":"2020","source":"Animals : an open access journal from MDPI","url":"https://pubmed.ncbi.nlm.nih.gov/32033330","citation_count":18,"is_preprint":false},{"pmid":"20007546","id":"PMC_20007546","title":"Retroviral insertional mutagenesis identifies Zeb2 activation as a novel leukemogenic collaborating event in CALM-AF10 transgenic mice.","date":"2009","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/20007546","citation_count":18,"is_preprint":false},{"pmid":"11135434","id":"PMC_11135434","title":"Molecular analysis of the genomic inversion and insertion of AF10 into MLL suggests a single-step event.","date":"2001","source":"Genes, chromosomes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/11135434","citation_count":17,"is_preprint":false},{"pmid":"33967269","id":"PMC_33967269","title":"Tip60 activates Hoxa9 and Meis1 expression through acetylation of H2A.Z, promoting MLL-AF10 and MLL-ENL acute myeloid leukemia.","date":"2021","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/33967269","citation_count":16,"is_preprint":false},{"pmid":"31022428","id":"PMC_31022428","title":"Acute myeloid leukemia driven by the CALM-AF10 fusion gene is dependent on BMI1.","date":"2019","source":"Experimental hematology","url":"https://pubmed.ncbi.nlm.nih.gov/31022428","citation_count":16,"is_preprint":false},{"pmid":"33690798","id":"PMC_33690798","title":"A JAK/STAT-mediated inflammatory signaling cascade drives oncogenesis in AF10-rearranged AML.","date":"2021","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/33690798","citation_count":15,"is_preprint":false},{"pmid":"34226546","id":"PMC_34226546","title":"The role of the PZP domain of AF10 in acute leukemia driven by AF10 translocations.","date":"2021","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/34226546","citation_count":14,"is_preprint":false},{"pmid":"11165485","id":"PMC_11165485","title":"The Drosophila homolog of human AF10/AF17 leukemia fusion genes (Dalf) encodes a zinc finger/leucine zipper nuclear protein required in the nervous system for maintaining EVE expression and normal growth.","date":"2001","source":"Mechanisms of development","url":"https://pubmed.ncbi.nlm.nih.gov/11165485","citation_count":14,"is_preprint":false},{"pmid":"9844603","id":"PMC_9844603","title":"Interstitial insertion of AF10 into the ALL1 gene in a case of infant acute lymphoblastic leukemia.","date":"1998","source":"Cancer genetics and cytogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/9844603","citation_count":14,"is_preprint":false},{"pmid":"17074587","id":"PMC_17074587","title":"A case of acute myelogenous leukemia with MLL-AF10 fusion caused by insertion of 5' MLL into 10p12, with concurrent 3' MLL deletion.","date":"2006","source":"Cancer genetics and cytogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/17074587","citation_count":14,"is_preprint":false},{"pmid":"34215314","id":"PMC_34215314","title":"AF10 (MLLT10) prevents somatic cell reprogramming through regulation of DOT1L-mediated H3K79 methylation.","date":"2021","source":"Epigenetics & chromatin","url":"https://pubmed.ncbi.nlm.nih.gov/34215314","citation_count":13,"is_preprint":false},{"pmid":"21706055","id":"PMC_21706055","title":"The clathrin-binding domain of CALM-AF10 alters the phenotype of myeloid neoplasms in mice.","date":"2011","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/21706055","citation_count":13,"is_preprint":false},{"pmid":"15262427","id":"PMC_15262427","title":"MLL-MLLT10 fusion in acute monoblastic leukemia: variant complex rearrangements and 11q proximal breakpoint heterogeneity.","date":"2004","source":"Cancer genetics and cytogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/15262427","citation_count":13,"is_preprint":false},{"pmid":"11417476","id":"PMC_11417476","title":"Identification and molecular characterisation of a CALM-AF10 fusion in acute megakaryoblastic leukaemia.","date":"2001","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/11417476","citation_count":13,"is_preprint":false},{"pmid":"9305618","id":"PMC_9305618","title":"A novel MLL-AF10 fusion mRNA variant in a patient with acute myeloid leukemia detected by a new asymmetric reverse transcription PCR method.","date":"1997","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/9305618","citation_count":13,"is_preprint":false},{"pmid":"23263989","id":"PMC_23263989","title":"The conserved PHD1-PHD2 domain of ZFP-1/AF10 is a discrete functional module essential for viability in Caenorhabditis elegans.","date":"2012","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/23263989","citation_count":12,"is_preprint":false},{"pmid":"16111539","id":"PMC_16111539","title":"MLL-MLLT10 fusion gene in pediatric acute megakaryoblastic leukemia.","date":"2005","source":"Leukemia research","url":"https://pubmed.ncbi.nlm.nih.gov/16111539","citation_count":12,"is_preprint":false},{"pmid":"11911106","id":"PMC_11911106","title":"Incidence of MLL rearrangement in acute myeloid leukemia, and a CALM-AF10 fusion in M4 type acute myeloblastic leukemia.","date":"2002","source":"Leukemia & lymphoma","url":"https://pubmed.ncbi.nlm.nih.gov/11911106","citation_count":12,"is_preprint":false},{"pmid":"16213369","id":"PMC_16213369","title":"Cryptic MLL-AF10 fusion caused by insertion of duplicated 5' part of MLL into 10p12 in acute leukemia: a case report.","date":"2005","source":"Cancer genetics and cytogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/16213369","citation_count":12,"is_preprint":false},{"pmid":"38306602","id":"PMC_38306602","title":"Structural variants involving MLLT10 fusion are associated with adverse outcomes in pediatric acute myeloid leukemia.","date":"2024","source":"Blood advances","url":"https://pubmed.ncbi.nlm.nih.gov/38306602","citation_count":11,"is_preprint":false},{"pmid":"28931923","id":"PMC_28931923","title":"Mllt10 knockout mouse model reveals critical role of Af10-dependent H3K79 methylation in midfacial development.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/28931923","citation_count":11,"is_preprint":false},{"pmid":"35702670","id":"PMC_35702670","title":"Pleiotropic MLLT10 variation confers risk of meningioma and estrogen-mediated cancers.","date":"2022","source":"Neuro-oncology advances","url":"https://pubmed.ncbi.nlm.nih.gov/35702670","citation_count":11,"is_preprint":false},{"pmid":"26686248","id":"PMC_26686248","title":"The target cell of transformation is distinct from the leukemia stem cell in murine CALM/AF10 leukemia models.","date":"2015","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/26686248","citation_count":11,"is_preprint":false},{"pmid":"17868029","id":"PMC_17868029","title":"AF10-dependent transcription is enhanced by its interaction with FLRG.","date":"2007","source":"Biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/17868029","citation_count":11,"is_preprint":false},{"pmid":"11106826","id":"PMC_11106826","title":"CALM-AF10 fusion gene in leukemias: simple and inversion-associated translocation (10;11).","date":"2000","source":"Cancer genetics and cytogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/11106826","citation_count":11,"is_preprint":false},{"pmid":"11187895","id":"PMC_11187895","title":"Protean clinical manifestations in children with leukemias containing MLL-AF10 fusion.","date":"2000","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/11187895","citation_count":11,"is_preprint":false},{"pmid":"37743097","id":"PMC_37743097","title":"Treatment outcomes of childhood PICALM::MLLT10 acute leukaemias.","date":"2023","source":"British journal of haematology","url":"https://pubmed.ncbi.nlm.nih.gov/37743097","citation_count":10,"is_preprint":false},{"pmid":"30159143","id":"PMC_30159143","title":"Dysregulated transcriptional networks in KMT2A- and MLLT10-rearranged T-ALL.","date":"2018","source":"Biomarker research","url":"https://pubmed.ncbi.nlm.nih.gov/30159143","citation_count":10,"is_preprint":false},{"pmid":"24397609","id":"PMC_24397609","title":"Nuclear export signal within CALM is necessary for CALM-AF10-induced leukemia.","date":"2014","source":"Cancer science","url":"https://pubmed.ncbi.nlm.nih.gov/24397609","citation_count":10,"is_preprint":false},{"pmid":"20445327","id":"PMC_20445327","title":"Clathrin assembly lymphoid myeloid leukemia-AF10-positive acute leukemias: a report of 2 cases with a review of the literature.","date":"2010","source":"The Korean journal of laboratory medicine","url":"https://pubmed.ncbi.nlm.nih.gov/20445327","citation_count":10,"is_preprint":false},{"pmid":"12555219","id":"PMC_12555219","title":"Monocytic leukemia with CALM/AF10 rearrangement showing mediastinal emphysema.","date":"2003","source":"American journal of hematology","url":"https://pubmed.ncbi.nlm.nih.gov/12555219","citation_count":10,"is_preprint":false},{"pmid":"19711340","id":"PMC_19711340","title":"MLL/AF10(OM-LZ)-immortalized cells expressed cytokines and induced host cell proliferation in a mouse bone marrow transplantation model.","date":"2010","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/19711340","citation_count":10,"is_preprint":false},{"pmid":"27859216","id":"PMC_27859216","title":"Cooperation of MLL/AF10(OM-LZ) with PTPN11 activating mutation induced monocytic leukemia with a shorter latency in a mouse bone marrow transplantation model.","date":"2016","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/27859216","citation_count":10,"is_preprint":false},{"pmid":"32569758","id":"PMC_32569758","title":"MLLT10 in benign and malignant hematopoiesis.","date":"2020","source":"Experimental hematology","url":"https://pubmed.ncbi.nlm.nih.gov/32569758","citation_count":9,"is_preprint":false},{"pmid":"30227397","id":"PMC_30227397","title":"Mediastinal Myeloid Sarcoma with TP53 Mutation Preceding Acute Myeloid Leukemia with a PICALM-MLLT10 Fusion Gene.","date":"2018","source":"Acta haematologica","url":"https://pubmed.ncbi.nlm.nih.gov/30227397","citation_count":9,"is_preprint":false},{"pmid":"37478796","id":"PMC_37478796","title":"Cryptic KMT2A/MLLT10 fusion detected by next-generation sequencing in a case of pediatric acute megakaryoblastic leukemia.","date":"2023","source":"Cancer genetics","url":"https://pubmed.ncbi.nlm.nih.gov/37478796","citation_count":8,"is_preprint":false},{"pmid":"32843425","id":"PMC_32843425","title":"Failure of tofacitinib to achieve an objective response in a DDX3X-MLLT10 T-lymphoblastic leukemia with activating JAK3 mutations.","date":"2020","source":"Cold Spring Harbor molecular case studies","url":"https://pubmed.ncbi.nlm.nih.gov/32843425","citation_count":8,"is_preprint":false},{"pmid":"37082953","id":"PMC_37082953","title":"The DOT1L-MLLT10 complex regulates male fertility and promotes histone removal during spermiogenesis.","date":"2023","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/37082953","citation_count":7,"is_preprint":false},{"pmid":"24558261","id":"PMC_24558261","title":"Neuronal migration is regulated by endogenous RNAi and chromatin-binding factor ZFP-1/AF10 in Caenorhabditis elegans.","date":"2014","source":"Genetics","url":"https://pubmed.ncbi.nlm.nih.gov/24558261","citation_count":7,"is_preprint":false},{"pmid":"24692230","id":"PMC_24692230","title":"Characterization and inhibition of AF10-mediated interaction.","date":"2014","source":"Journal of peptide science : an official publication of the European Peptide Society","url":"https://pubmed.ncbi.nlm.nih.gov/24692230","citation_count":7,"is_preprint":false},{"pmid":"31739126","id":"PMC_31739126","title":"Molecular and phenotypic characterization of an early T-cell precursor acute lymphoblastic lymphoma harboring PICALM-MLLT10 fusion with aberrant expression of B-cell antigens.","date":"2019","source":"Cancer genetics","url":"https://pubmed.ncbi.nlm.nih.gov/31739126","citation_count":7,"is_preprint":false},{"pmid":"36158701","id":"PMC_36158701","title":"Broad genomic workup including optical genome mapping uncovers a DDX3X: MLLT10 gene fusion in acute myeloid leukemia.","date":"2022","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/36158701","citation_count":7,"is_preprint":false},{"pmid":"38971327","id":"PMC_38971327","title":"Clinicopathologic features and outcomes of acute leukemia harboring PICALM::MLLT10 fusion.","date":"2024","source":"Human pathology","url":"https://pubmed.ncbi.nlm.nih.gov/38971327","citation_count":6,"is_preprint":false},{"pmid":"38895061","id":"PMC_38895061","title":"PICALM::MLLT10 may indicate a new subgroup of acute leukemias with miscellaneous immunophenotype and poor initial treatment response but showing sensitivity to venetoclax.","date":"2024","source":"EJHaem","url":"https://pubmed.ncbi.nlm.nih.gov/38895061","citation_count":6,"is_preprint":false},{"pmid":"8373639","id":"PMC_8373639","title":"Interferon-alpha and dexamethasone effect on AF10 myeloma cell line sialytransferase activity.","date":"1993","source":"Biochemical medicine and metabolic biology","url":"https://pubmed.ncbi.nlm.nih.gov/8373639","citation_count":6,"is_preprint":false},{"pmid":"25193880","id":"PMC_25193880","title":"Effects of iron depletion on CALM-AF10 leukemias.","date":"2014","source":"Experimental hematology","url":"https://pubmed.ncbi.nlm.nih.gov/25193880","citation_count":6,"is_preprint":false},{"pmid":"9762409","id":"PMC_9762409","title":"A human B cell line AF10 expressing HIL-17.","date":"1998","source":"Biochemistry and molecular biology international","url":"https://pubmed.ncbi.nlm.nih.gov/9762409","citation_count":6,"is_preprint":false},{"pmid":"37995701","id":"PMC_37995701","title":"DOT1L interaction partner AF10 controls patterning of H3K79 methylation and RNA polymerase II to maintain cell identity.","date":"2023","source":"Stem cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/37995701","citation_count":5,"is_preprint":false},{"pmid":"28781666","id":"PMC_28781666","title":"Molecular studies reveal MLL-MLLT10/AF10 and ARID5B-MLL gene fusions displaced in a case of infantile acute lymphoblastic leukemia with complex karyotype.","date":"2017","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/28781666","citation_count":5,"is_preprint":false},{"pmid":"8946209","id":"PMC_8946209","title":"Gene BR140, which is related to AF10 and AF17, maps to chromosome band 3p25.","date":"1996","source":"Genes, chromosomes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/8946209","citation_count":5,"is_preprint":false},{"pmid":"36774707","id":"PMC_36774707","title":"Unexpected appearance of KMT2A::MLLT10 fusion transcript in acute myeloid leukemia with t(5;11)(q31;q23.3).","date":"2023","source":"Cancer genetics","url":"https://pubmed.ncbi.nlm.nih.gov/36774707","citation_count":5,"is_preprint":false},{"pmid":"15258852","id":"PMC_15258852","title":"Molecular genetics of the Alhambra (Drosophila AF10) complex locus of Drosophila.","date":"2004","source":"Molecular genetics and genomics : MGG","url":"https://pubmed.ncbi.nlm.nih.gov/15258852","citation_count":5,"is_preprint":false},{"pmid":"27510896","id":"PMC_27510896","title":"Transient spontaneous remission in congenital MLL-AF10 rearranged acute myeloid leukemia presenting with cardiorespiratory failure and meconium ileus.","date":"2016","source":"Molecular and cellular pediatrics","url":"https://pubmed.ncbi.nlm.nih.gov/27510896","citation_count":5,"is_preprint":false},{"pmid":"16518848","id":"PMC_16518848","title":"Acute monocytic leukemia with coexpression of minor BCR-ABL1 and PICALM-MLLT10 fusion genes along with overexpression of HOXA9.","date":"2006","source":"Genes, chromosomes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/16518848","citation_count":5,"is_preprint":false},{"pmid":"25435396","id":"PMC_25435396","title":"MLLT10 and IL3 rearrangement together with a complex four-way translocation and trisomy 4 in a patient with early T-cell precursor acute lymphoblastic leukemia: A case report.","date":"2014","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/25435396","citation_count":5,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":48053,"output_tokens":7716,"usd":0.129949,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":17170,"output_tokens":5635,"usd":0.113363,"stage2_stop_reason":"end_turn"},"total_usd":0.243312,"stage1_batch_id":"msgbatch_01BbTCMyrRPyhtEq6b1MdwCN","stage2_batch_id":"msgbatch_011dEed5bbxYvTAaQWKJevxu","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1995,\n      \"finding\": \"AF10 protein contains a conserved cysteine-rich LAP/PHD zinc finger motif (now called LAP finger) that is proposed to be a DNA-binding domain; the LAP domain consensus sequence was defined by homology across 25+ proteins and the domain is disrupted in leukemia-associated chromosomal translocations.\",\n      \"method\": \"Sequence homology analysis and structural characterization of zinc finger motif\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 / Weak — computational/sequence-based prediction only; no direct binding experiment performed on AF10 specifically\",\n      \"pmids\": [\"7568208\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"In the t(10;11)(p12;q23) translocation causing AML, the leucine zipper motif of AF10 is consistently fused onto the N-terminal region of MLL/HRX, establishing that the leucine zipper of AF10 is a critical functional element juxtaposed onto MLL in all examined cases.\",\n      \"method\": \"Southern analysis and RT-PCR sequencing of leukemia patient samples\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple patient samples with consistent molecular breakpoint analysis, single lab\",\n      \"pmids\": [\"7662954\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"AF10 contains an extended LAP/PHD finger domain that mediates homo-oligomerization of recombinant AF10 protein; AF10 also binds cruciform DNA via an AT-hook motif and is localized to the nucleus by a bipartite nuclear localization signal in its N-terminal region.\",\n      \"method\": \"Biochemical analysis of recombinant AF10 protein; GST pulldown for oligomerization; DNA-binding assay; subcellular fractionation\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct biochemical reconstitution with recombinant protein, multiple orthogonal methods, single lab\",\n      \"pmids\": [\"10860745\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"AF10 interacts with the Drosophila heterochromatin protein HP1 (in vitro and in vivo), and the Drosophila AF10 homolog dAF10 functions in heterochromatin-dependent gene silencing (position effect variegation), placing it in the HP1-dependent silencing pathway.\",\n      \"method\": \"In vitro binding assay and co-immunoprecipitation (in vivo); genetic analysis of position effect variegation in Drosophila\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal in vitro and in vivo binding plus genetic functional assay, single lab, ortholog\",\n      \"pmids\": [\"11266362\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"The AF10 leucine zipper domain interacts with GAS41 (a glioblastoma amplified gene product homologous to yeast ANC1); GAS41 in turn co-immunoprecipitates with INI1 (SNF5/SWI-SNF component), and INI1 is present in AF10 immunoprecipitates, placing AF10 in proximity to the SWI/SNF chromatin remodeling complex.\",\n      \"method\": \"Yeast two-hybrid screen; co-immunoprecipitation (in vivo confirmation)\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — yeast 2-hybrid plus reciprocal co-IP confirmation, single lab\",\n      \"pmids\": [\"11756182\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"The AF10 leucine zipper (comprising two adjacent alpha-helical domains, 82 aa) is necessary and sufficient for MLL-AF10-mediated myeloid immortalization and leukemic transformation; deletion of the 29-aa leucine zipper within this region completely abrogates transforming activity; the same minimal domain also confers transcriptional activation when fused to MLL or GAL4 DNA-binding domains.\",\n      \"method\": \"Structure-function analysis with retroviral transduction of deletion mutants into primary murine myeloid progenitors; serial replating assays; in vivo leukemia model; GAL4 transcriptional reporter assay\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — multiple deletion mutants tested, in vitro and in vivo transformation assays, transcriptional readout, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"11986236\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"AF10 physically interacts with the synovial sarcoma-associated protein SYT; the interaction was mapped to the N-terminal region of SYT and a C-terminal region of AF10 outside known functional domains; co-localization confirmed in transfected cells.\",\n      \"method\": \"Yeast two-hybrid screen; co-immunoprecipitation of endogenous and epitope-tagged proteins; co-localization by immunofluorescence; sequential deletion mapping\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP plus localization and domain mapping, single lab\",\n      \"pmids\": [\"11423977\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"The Drosophila AF10 homolog Alhambra (ALH) full-length protein has no activity on Polycomb group-responsive elements (PREs), but overexpression of the isolated leucine zipper domain activates several PREs; the PHD domain within the full-length protein inhibits the PRE-deregulating activity of the leucine zipper; this PRE deregulation is conserved in the human AF10 leucine zipper domain expressed in Drosophila.\",\n      \"method\": \"Drosophila genetics; PRE reporter assays; domain deletion/overexpression analysis in flies\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with domain-specific alleles in Drosophila ortholog, single lab\",\n      \"pmids\": [\"12482966\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"CALM-AF10 fusion protein interacts with the H3K79 methyltransferase hDOT1L through the AF10 moiety; hDOT1L prevents nuclear export of CALM-AF10 and upregulates Hoxa5 through H3K79 methylation, contributing to leukemic transformation.\",\n      \"method\": \"Co-immunoprecipitation; ChIP for H3K79 methylation at Hoxa5 locus; retroviral bone marrow transformation assay; nuclear export assay\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP, ChIP showing H3K79me at specific locus, functional transformation assay, multiple orthogonal methods, single lab\",\n      \"pmids\": [\"16921363\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"The CALM-AF10 fusion protein alters subcellular localization of the lymphoid transcription factor Ikaros by coexpression; the AF10 leucine zipper domain is required for the AF10-Ikaros interaction; the transcriptional repressor activity of Ikaros is reduced by AF10.\",\n      \"method\": \"Yeast two-hybrid screen; GST pulldown; co-immunoprecipitation; immunofluorescence co-localization; transcriptional reporter assay\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — GST pulldown plus co-IP plus localization plus reporter assay, single lab\",\n      \"pmids\": [\"18037964\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"A novel CALM-interacting protein, CATS, increases the nuclear (and specifically nucleolar) localization of both CALM and the leukemogenic CALM/AF10 fusion protein; the CATS interaction domain was mapped to aa 221-335 of CALM.\",\n      \"method\": \"Yeast two-hybrid screen; GST pulldown; co-immunoprecipitation; co-localization by fluorescence microscopy; domain mapping\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal binding methods plus functional localization effect, single lab\",\n      \"pmids\": [\"16491119\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"The CALM-AF10 fusion protein disrupts the normal AF10-mediated association of hDOT1L with chromatin, causing a global reduction of H3K79 methylation; cells with reduced H3K79 methylation show increased sensitivity to gamma-irradiation and chromosomal instability.\",\n      \"method\": \"ChIP analysis of H3K79 methylation; co-immunoprecipitation; gamma-irradiation sensitivity assays; cytogenetic analysis of leukemia patient samples\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP plus co-IP plus functional cellular assay, single lab\",\n      \"pmids\": [\"19443658\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"MLLT10/AF10 and DOT1L are TCF4/β-catenin interactors in intestinal crypts; the AF10-DOT1L complex is recruited to Wnt target genes in a β-catenin-dependent manner and deposits H3K79 methylation over their coding regions; depletion of MLLT10/AF10 selectively impairs Wnt target gene expression and intestinal proliferation.\",\n      \"method\": \"Proteomics/mass spectrometry of TCF4/β-catenin complex; ChIP-H3K79me on Wnt target genes; siRNA knockdown with expression array; zebrafish morpholino experiments; genetic epistasis with apc-mutant zebrafish\",\n      \"journal\": \"PLoS biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — proteomics identification + ChIP + genetic epistasis + multiple model organisms + expression arrays, replicated across systems\",\n      \"pmids\": [\"21103407\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"For CALM-AF10 leukemia transformation, the clathrin-binding domain of CALM (C-terminal 248 aa) and the octapeptide motif-leucine zipper (OM-LZ) domain of AF10 are the minimal sufficient domains; this 'minimal fusion' retains aberrant Hoxa cluster upregulation characteristic of full-length CALM-AF10.\",\n      \"method\": \"Structure-function analysis with CALM and AF10 domain deletion mutants; colony-forming assays; in vivo mouse leukemia model; gene expression analysis\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — systematic domain deletion with in vitro and in vivo transformation assays, molecular phenotype confirmed, single lab\",\n      \"pmids\": [\"21681188\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Full-length CALM-AF10 localizes to the nucleus (not cytoplasm) in leukemia cells and has a propensity to homo-oligomerize as shown by FRET analysis; CALM-AF10 suppresses H3K79 methylation regardless of the presence of clathrin.\",\n      \"method\": \"Fluorescence microscopy; FRET analysis; ChIP for H3K79 methylation; mouse leukemia model with domain deletion constructs\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — FRET for oligomerization, ChIP for epigenetic mark, localization by imaging, single lab\",\n      \"pmids\": [\"21706055\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"MLL-AF10 and CALM-AF10 mediated transformation is dependent on the H3K79 methyltransferase DOT1L; conditional Dot1l knockout abolishes in vitro transformation and in vivo leukemia initiation/maintenance; pharmacological DOT1L inhibition (EPZ004777) suppresses Hoxa cluster genes and Meis1, and selectively impairs proliferation of MLL-AF10 and CALM-AF10 transformed cells.\",\n      \"method\": \"Conditional Dot1l knockout mouse model; in vitro colony transformation assay; in vivo leukemia model; DOT1L inhibitor (EPZ004777) treatment; gene expression analysis\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — genetic knockout combined with pharmacological inhibition, in vitro and in vivo models, replicated across two fusion oncoproteins\",\n      \"pmids\": [\"23138183\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"The PHD1-PHD2 module of the C. elegans AF10 ortholog ZFP-1 is essential for viability; the first PHD finger contributes to preferential binding of the PHD1-PHD2 domain to H3K4-methylated histone H3 tails; ZFP-1 occupancy genome-wide peaks at H3K4me-enriched promoters of active genes, and H3K4 methylation is required for ZFP-1 localization to promoters in the embryo.\",\n      \"method\": \"Genetic analysis (C. elegans deletion mutants); biochemical histone-tail binding assays; ChIP-seq for ZFP-1 and H3K4me marks\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — genetic viability assay, biochemical histone binding, genome-wide ChIP-seq, multiple orthogonal methods in the ortholog\",\n      \"pmids\": [\"23263989\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"The C. elegans AF10 homolog ZFP-1 and its interacting partner DOT-1.1 globally reduce RNA Pol II transcription on essential widely expressed genes; the ZFP-1/DOT-1.1 complex increases H3K79 methylation and decreases H2B monoubiquitination (which promotes transcription), thereby negatively modulating Pol II elongation.\",\n      \"method\": \"Genomic (ChIP-seq, RNA-seq) and biochemical approaches; genetic knockdown; Pol II pausing analysis during development and stress response\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genome-wide ChIP-seq, RNA-seq, biochemical co-complex analysis, multiple orthogonal methods in C. elegans ortholog\",\n      \"pmids\": [\"23806335\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"CALM contains a CRM1-dependent nuclear export signal (NES) that mediates cytoplasmic localization of CALM-AF10 and is necessary for CALM-AF10-dependent transformation; NES motifs from heterologous proteins fused in-frame with AF10 are sufficient to immortalize murine hematopoietic progenitors; the CALM NES is essential for Hoxa gene upregulation and aberrant H3K79 methylation by CALM-AF10, possibly through mislocalization of DOT1L.\",\n      \"method\": \"NES mutational analysis; retroviral transformation assay; ChIP for H3K79me and HOXA gene expression; Leptomycin B (CRM1 inhibitor) treatment; heterologous NES fusion constructs\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — systematic mutagenesis, functional transformation assay, pharmacological inhibition, ChIP validation, multiple orthogonal methods\",\n      \"pmids\": [\"23487024\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"CRM1 physically localizes to HOXA loci and recruits CALM-AF10 to HOXA chromatin through the CALM NES-CRM1 interaction; genetic and pharmacological inhibition of CALM-CRM1 interaction prevents CALM-AF10 enrichment at HOXA chromatin and immediately abolishes HOXA transcription; CRM1 thus has a novel chromatin-binding and transcription factor-recruiting function.\",\n      \"method\": \"ChIP for CRM1 and CALM-AF10 at HOXA loci; genetic CALM NES mutants; CRM1 inhibitor treatment; transcriptional analysis\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — ChIP showing CRM1 and CALM-AF10 co-occupancy, genetic NES mutants, pharmacological inhibition, multiple orthogonal methods\",\n      \"pmids\": [\"25027513\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"The AF10 PZP domain (PHD finger-Zn knuckle-PHD finger module) reads unmodified H3K27; structural studies reveal that H3 binding triggers rearrangement of the PZP module to form an H3(22-27)-accommodating channel where unmodified H3K27 is encaged in a hydrogen-bond acceptor-lined cavity; H3K27 modification abrogates this interaction. In cells, PZP recognition of H3 is required for H3K79 dimethylation, expression of DOT1L-target genes, and proliferation of DOT1L-addicted leukemic cells.\",\n      \"method\": \"Crystal structure of PZP-H3 complex; mutagenesis of binding interface; histone peptide pulldowns; ChIP for H3K79me2 in cells with PZP mutants; cell proliferation assays\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure plus mutagenesis plus ChIP in cells plus functional proliferation assay, multiple orthogonal methods in single rigorous study\",\n      \"pmids\": [\"26439302\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Mllt10 knockout mice exhibit midline facial cleft associated with reduced H3K79 methylation in nasal processes; AP2α expression is directly regulated by Af10-dependent H3K79me at its locus, and is specifically reduced in nasal processes of Mllt10-KO embryos; pharmacological suppression of H3K79me completely phenocopies Mllt10-KO.\",\n      \"method\": \"Mllt10 knockout mouse; ChIP for H3K79me at AP2α locus in nasal processes; DOT1L inhibitor treatment phenocopy experiment; gene expression analysis\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO with defined molecular phenotype, ChIP at specific locus, pharmacological phenocopy, multiple orthogonal approaches\",\n      \"pmids\": [\"28931923\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Crystal structures of both apo AF10 OM-LZ and its complex with the coiled-coil domain of DOT1L were solved; disruption of the DOT1L-AF10 interface abrogates MLL-AF10-associated leukemic transformation; zinc stabilizes the DOT1L-AF10 complex and may regulate HOXA gene expression.\",\n      \"method\": \"X-ray crystallography of AF10 OM-LZ alone and in complex with DOT1L coiled-coil domain; interface mutagenesis; leukemic transformation assay\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure plus mutagenesis of interface plus functional leukemia transformation assay in single study\",\n      \"pmids\": [\"29563185\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"AF10 fusions (PICALM-AF10 and MLL-AF10) directly recruit JAK1 kinase and activate a JAK/STAT-mediated inflammatory signaling cascade; genetic Jak1 deletion or pharmacological JAK/STAT inhibition elicits potent anti-oncogenic effects in mouse and human AF10-fusion AML models.\",\n      \"method\": \"Inducible mouse AML models; proteomics/protein interactome analysis; genetic Jak1 conditional knockout; pharmacological JAK/STAT inhibition; transcriptomic and epigenomic profiling\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — proteomic identification of JAK1 as direct interactor, genetic KO, pharmacological inhibition, multiple AF10 fusions tested, multiple orthogonal approaches\",\n      \"pmids\": [\"33690798\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"The AF10 PZP domain (AF10PZP) binds the nucleosome core particle through multivalent contacts with the histone H3 tail and DNA; AF10PZP associates with active chromatin marks and discriminates against H3K27me3; disruption of AF10PZP function in CALM-AF10 drives transformation, while incorporation of functional AF10PZP into CALM-AF10 prevents transformation, promotes nuclear localization, and downregulates Hoxa genes.\",\n      \"method\": \"Crystallography of AF10PZP-nucleosome complex; biochemical binding assays; mutagenesis; bone marrow transformation assays in vitro and in vivo; ChIP; gene expression analysis\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure, biochemical assays, mutagenesis, in vitro and in vivo functional transformation assays, ChIP, multiple orthogonal methods\",\n      \"pmids\": [\"34226546\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"AF10 (MLLT10) is required for higher-order H3K79 methylation (H3K79me2/me3) in somatic cells; suppression of AF10 via RNAi or CRISPR/Cas9 significantly increases somatic cell reprogramming efficiency; re-expression of wild-type AF10 but not a DOT1L-binding-impaired AF10 mutant rescues H3K79 methylation and reduces reprogramming efficiency, establishing AF10 as a barrier to reprogramming through its regulation of DOT1L-dependent H3K79 methylation.\",\n      \"method\": \"Proximity-based labeling proteomics (BioID); RNAi and CRISPR/Cas9 knockdown; reprogramming efficiency assay; rescue with wild-type vs. DOT1L-binding mutant AF10; H3K79me western blot; transcriptomic analysis\",\n      \"journal\": \"Epigenetics & chromatin\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — proteomics identification, genetic KO, structure-function rescue with DOT1L-binding mutant, multiple orthogonal approaches in single study\",\n      \"pmids\": [\"34215314\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Tip60 histone acetyltransferase is recruited by MLL-AF10 to the Hoxa9 locus where it acetylates H2A.Z to promote Hoxa9 gene expression; conditional deletion of Tip60 prevents development of MLL-AF10 leukemia.\",\n      \"method\": \"ChIP showing Tip60 recruitment to Hoxa9 by MLL-AF10; H2A.Z acetylation assay; conditional Tip60 knockout; leukemia development assay in mice\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — ChIP, histone modification assay, genetic conditional KO with functional leukemia phenotype, multiple orthogonal methods\",\n      \"pmids\": [\"33967269\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"DOT1L associates with MLLT10 in testis; DOT1L and MLLT10 co-localize to sex chromatin in meiotic and post-meiotic germ cells in an inter-dependent manner; both DOT1L and MLLT10 are essential for H3K79me2 in germ cells and for histone-to-protamine replacement during spermiogenesis; loss of either DOT1L or MLLT10 results in histone retention in sperm and male subfertility.\",\n      \"method\": \"Co-immunoprecipitation (DOT1L-MLLT10 in testis); immunofluorescence co-localization in germ cells; Mllt10 conditional knockout mouse; H3K79me2 ChIP/immunostaining; sperm histone retention assay\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — co-IP, co-localization, conditional KO with defined epigenetic and developmental phenotype, multiple orthogonal methods\",\n      \"pmids\": [\"37082953\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"AF10 deletion evicts H3K79me2/3 from gene bodies and redistributes H3K79me1 to transcription start sites; AF10 loss also redistributes RNA Pol II to a pattern characteristic of pluripotency at highly expressed housekeeping genes, facilitating iPSC formation without major steady-state transcriptional changes; this reveals that AF10 controls the patterning of H3K79 methylation orders to maintain cell identity.\",\n      \"method\": \"AF10 genetic deletion (CRISPR); CUT&RUN/ChIP for H3K79me1/me2/me3; RNA Pol II ChIP-seq; reprogramming efficiency assay; DOT1L chemical inhibition as comparison\",\n      \"journal\": \"Stem cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO, genome-wide ChIP-seq for multiple histone marks and Pol II, reprogramming assay, chemical inhibition orthogonal comparison\",\n      \"pmids\": [\"37995701\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"The nuclear form of FLRG (follistatin-related gene) interacts with AF10 via the N-terminal PHD-containing region of AF10; FLRG enhances AF10 homo-oligomerization and augments the transcriptional activation properties of AF10 in reporter assays.\",\n      \"method\": \"Yeast two-hybrid screen; far-Western blot; co-immunoprecipitation; transcriptional reporter (Gal4-AF10 fusion); domain mapping\",\n      \"journal\": \"Biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple binding methods (Y2H, far-Western, co-IP) plus functional reporter, single lab\",\n      \"pmids\": [\"17868029\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MLLT10/AF10 is a transcriptional co-regulator that functions primarily as a cofactor for the H3K79 methyltransferase DOT1L: AF10 binds DOT1L through its octapeptide motif-leucine zipper (OM-LZ) domain (structurally resolved by X-ray crystallography), while its N-terminal PZP domain (PHD-Zn knuckle-PHD) reads unmodified H3K27 on nucleosomes to target DOT1L-dependent H3K79 dimethylation and trimethylation to gene bodies of active genes; AF10 also homo-oligomerizes via its extended LAP/PHD finger, binds chromatin via an AT-hook, localizes to the nucleus via a bipartite NLS, and associates with the SWI/SNF complex through GAS41 and INI1; in development, AF10-dependent H3K79 methylation regulates AP2α expression in nasal processes and histone-to-protamine replacement during spermiogenesis; in Wnt signaling, AF10-DOT1L is recruited by TCF4/β-catenin to deposit H3K79me over Wnt target gene coding regions; in leukemia, AF10 fusion proteins (MLL-AF10, CALM-AF10) mislocalize or misdirect DOT1L to HOXA loci, driving aberrant H3K79 methylation and HOXA cluster gene upregulation, and also directly recruit JAK1 to activate JAK/STAT inflammatory signaling.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"MLLT10/AF10 is a nuclear chromatin-associated cofactor that targets the H3K79 methyltransferase DOT1L to active chromatin, governing transcriptional programs in development, stem cell identity, and leukemia [#15, #20, #28]. AF10 binds DOT1L directly through its octapeptide motif-leucine zipper (OM-LZ) domain, an interaction resolved by crystallography and stabilized by zinc, and disruption of this interface abolishes MLL-AF10 leukemic transformation [#22]. Its N-terminal PZP module (PHD-Zn knuckle-PHD) engages the nucleosome through multivalent contacts with unmodified H3 (specifically unmethylated H3K27) and DNA, discriminating against repressive H3K27me3, thereby directing DOT1L-dependent H3K79 di- and trimethylation to gene bodies of active genes; PZP recognition is required for H3K79me2 deposition, DOT1L-target gene expression, and proliferation of DOT1L-addicted cells [#20, #24]. AF10 controls the higher-order patterning of H3K79 methylation across the genome and the redistribution of RNA Pol II, acting as a barrier to somatic cell reprogramming that maintains cell identity [#25, #28]. Through this DOT1L axis, AF10 directly regulates AP2\\u03b1 expression and midline facial development [#21] and drives histone-to-protamine replacement during spermiogenesis, where DOT1L and MLLT10 co-localize to sex chromatin and are mutually required for germ-cell H3K79me2 [#27]. In Wnt signaling, the AF10-DOT1L complex is recruited by TCF4/\\u03b2-catenin to deposit H3K79 methylation over Wnt target gene coding regions, supporting intestinal proliferation [#12]. In leukemia, AF10 fusion oncoproteins (MLL-AF10, CALM/PICALM-AF10) misdirect DOT1L to drive aberrant H3K79 methylation and HOXA cluster upregulation; the minimal transforming unit is the AF10 OM-LZ together with the CALM clathrin-binding/NES region, with the CALM NES recruiting CRM1 to HOXA chromatin [#13, #15, #18, #19]. AF10 fusions additionally recruit Tip60 to acetylate H2A.Z at Hoxa9 and directly recruit JAK1 to activate JAK/STAT inflammatory signaling, both of which are required for leukemogenesis [#23, #26].\",\n  \"teleology\": [\n    {\n      \"year\": 1995,\n      \"claim\": \"Established AF10's domain architecture and its central role in leukemia, defining the LAP/PHD zinc finger and identifying the leucine zipper as the element consistently fused to MLL in t(10;11) AML.\",\n      \"evidence\": \"Sequence homology analysis of zinc finger motifs and breakpoint mapping in AML patient samples\",\n      \"pmids\": [\"7568208\", \"7662954\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"LAP/PHD DNA-binding was predicted, not demonstrated\", \"no mechanistic link to a methyltransferase yet\", \"functional consequence of the leucine zipper fusion unresolved\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Defined the biochemical properties of AF10 domains, showing the extended LAP/PHD finger drives homo-oligomerization, an AT-hook binds cruciform DNA, and a bipartite NLS directs nuclear localization.\",\n      \"evidence\": \"Biochemical reconstitution with recombinant AF10, GST pulldown, DNA-binding assay, and subcellular fractionation\",\n      \"pmids\": [\"10860745\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"physiological relevance of cruciform DNA binding unclear\", \"oligomerization partners in cells not defined\", \"no chromatin target identified\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Linked AF10 to chromatin silencing and other binding partners, connecting it to HP1-dependent heterochromatin in Drosophila and identifying SYT and (later) FLRG as interactors.\",\n      \"evidence\": \"In vitro/in vivo binding, co-IP, position-effect-variegation genetics in Drosophila, and yeast two-hybrid with domain mapping\",\n      \"pmids\": [\"11266362\", \"11423977\", \"17868029\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"mechanistic role of HP1, SYT, FLRG interactions in mammalian AF10 function unestablished\", \"ortholog-based silencing data may not transfer directly\", \"FLRG/SYT not connected to the DOT1L axis\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Showed the AF10 leucine zipper is necessary and sufficient for MLL-AF10 transformation and confers transcriptional activation, and placed AF10 in proximity to SWI/SNF via GAS41 and INI1.\",\n      \"evidence\": \"Deletion-mutant retroviral transformation in murine progenitors, serial replating, GAL4 reporter assays, and yeast two-hybrid/co-IP\",\n      \"pmids\": [\"11986236\", \"11756182\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"the leucine-zipper effector partner was not yet identified\", \"relationship between SWI/SNF association and transformation unclear\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Identified DOT1L as the key AF10 effector in leukemia, showing CALM-AF10 binds hDOT1L through the AF10 moiety to drive H3K79 methylation and Hoxa upregulation.\",\n      \"evidence\": \"Co-IP, ChIP for H3K79me at Hoxa5, nuclear export assay, and retroviral transformation; plus Ikaros and CATS interaction mapping\",\n      \"pmids\": [\"16921363\", \"18037964\", \"16491119\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"how AF10 normally targets DOT1L to chromatin not yet defined\", \"global versus local H3K79me effects unresolved\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Revealed that CALM-AF10 disrupts normal AF10-mediated DOT1L-chromatin association, causing global H3K79me loss, genomic instability, and irradiation sensitivity.\",\n      \"evidence\": \"ChIP for H3K79me, co-IP, gamma-irradiation sensitivity assays, and cytogenetics of patient samples\",\n      \"pmids\": [\"19443658\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"reconciling global H3K79me loss with locus-specific HOXA gain unresolved\", \"molecular basis of mistargeting not yet structural\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Defined a physiological developmental role by showing the AF10-DOT1L complex is recruited by TCF4/\\u03b2-catenin to deposit H3K79me over Wnt target gene coding regions and drive intestinal proliferation.\",\n      \"evidence\": \"TCF4/\\u03b2-catenin proteomics, ChIP-H3K79me, siRNA expression arrays, and zebrafish epistasis with apc mutants\",\n      \"pmids\": [\"21103407\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"direct contacts between AF10 and the TCF4/\\u03b2-catenin module not mapped\", \"selectivity for Wnt over other active genes unexplained\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Defined the minimal CALM-AF10 transforming unit (CALM clathrin-binding domain plus AF10 OM-LZ) and showed full-length CALM-AF10 localizes to the nucleus, homo-oligomerizes, and suppresses H3K79me independent of clathrin.\",\n      \"evidence\": \"Domain-deletion colony and mouse leukemia assays, FRET oligomerization analysis, and ChIP for H3K79me\",\n      \"pmids\": [\"21681188\", \"21706055\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"mechanism connecting oligomerization to H3K79me dysregulation unclear\", \"role of CALM moiety in DOT1L mistargeting not yet defined\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Established DOT1L as the genetic and pharmacological dependency of AF10-fusion leukemia and characterized AF10/DOT1L as a repressive elongation modulator in the ortholog.\",\n      \"evidence\": \"Conditional Dot1l knockout, EPZ004777 inhibition, and in vitro/in vivo leukemia models; plus C. elegans ZFP-1/DOT-1.1 ChIP-seq/RNA-seq\",\n      \"pmids\": [\"23138183\", \"23806335\", \"23263989\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"ortholog ZFP-1 repressive role contrasts with activating mammalian role, leaving directionality context-dependent\", \"PHD-module H3K4me reading shown in ortholog, not yet human\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Resolved how CALM-AF10 reaches HOXA chromatin, showing the CALM CRM1-dependent NES is required for transformation and recruits CRM1, which itself occupies HOXA loci and brings in CALM-AF10.\",\n      \"evidence\": \"NES mutagenesis, heterologous NES fusions, Leptomycin B treatment, and ChIP for CRM1 and CALM-AF10 at HOXA\",\n      \"pmids\": [\"23487024\", \"25027513\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"how CRM1 selects HOXA chromatin not defined\", \"generalizability beyond CALM-AF10 to MLL-AF10 unclear\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Provided the structural basis for AF10 chromatin targeting, showing the PZP module reads unmodified H3K27 to license DOT1L-dependent H3K79me2 and DOT1L-target gene expression.\",\n      \"evidence\": \"Crystal structure of PZP-H3 complex, interface mutagenesis, histone peptide pulldowns, and ChIP/proliferation assays in cells\",\n      \"pmids\": [\"26439302\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"nucleosomal (versus peptide) engagement not yet resolved at this stage\", \"interplay between PZP reading and OM-LZ DOT1L binding not integrated\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Demonstrated a non-leukemic developmental requirement, with Mllt10 knockout causing midline facial cleft through loss of Af10-dependent H3K79me at the AP2\\u03b1 locus.\",\n      \"evidence\": \"Mllt10 knockout mouse, ChIP for H3K79me at AP2\\u03b1 in nasal processes, and DOT1L-inhibitor phenocopy\",\n      \"pmids\": [\"28931923\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"other AF10-dependent developmental target genes not catalogued\", \"tissue-specific targeting mechanism unexplained\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Provided atomic detail of the AF10-DOT1L interface, solving the OM-LZ/DOT1L coiled-coil complex, showing zinc stabilization, and proving the interface is required for MLL-AF10 transformation.\",\n      \"evidence\": \"X-ray crystallography of AF10 OM-LZ alone and with DOT1L coiled-coil, interface mutagenesis, and leukemic transformation assay\",\n      \"pmids\": [\"29563185\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"functional role of zinc regulation in normal cells not defined\", \"whether the interface is druggable in leukemia untested here\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Integrated nucleosome-level PZP recognition, oncogenic JAK/STAT and Tip60 axes, and a reprogramming-barrier role, establishing AF10 as a multivalent chromatin reader controlling H3K79me-dependent cell identity and additional leukemogenic signaling.\",\n      \"evidence\": \"PZP-nucleosome crystallography with transformation rescue, BioID proteomics with reprogramming assays, and proteomics/genetic-pharmacological dissection of JAK1 and Tip60 in AML models\",\n      \"pmids\": [\"34226546\", \"34215314\", \"33690798\", \"33967269\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"how JAK1 recruitment relates mechanistically to H3K79me deposition unclear\", \"whether Tip60/H2A.Z acetylation acts upstream or in parallel to DOT1L unresolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Showed AF10 governs the higher-order patterning of H3K79 methylation and Pol II distribution to maintain cell identity and, in testis, partners with DOT1L for histone-to-protamine replacement during spermiogenesis.\",\n      \"evidence\": \"AF10 CRISPR deletion with CUT&RUN/ChIP-seq for H3K79me1/2/3 and Pol II plus reprogramming assays; and testis co-IP, co-localization, and Mllt10 conditional knockout with sperm histone-retention readout\",\n      \"pmids\": [\"37995701\", \"37082953\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"how AF10 selects which H3K79 methylation order to deposit at a given locus unknown\", \"mechanism coupling H3K79me patterning to Pol II redistribution undefined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How AF10's two chromatin-engaging modules (PZP nucleosome reading and OM-LZ DOT1L binding) are coordinated to choose specific loci and to set the order of H3K79 methylation, and how the non-DOT1L oncogenic activities (JAK1, Tip60, CRM1) are mechanistically integrated, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"no unified model linking PZP/OM-LZ engagement to locus selection\", \"mechanism setting H3K79me1 vs me2/me3 at a locus unknown\", \"integration of JAK/STAT, Tip60, and CRM1 axes with the DOT1L methyltransferase axis undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0042393\", \"supporting_discovery_ids\": [20, 24, 16]},\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [2, 24]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [5, 12, 29]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [20, 22, 24]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [2, 14]},\n      {\"term_id\": \"GO:0000228\", \"supporting_discovery_ids\": [20, 24, 28]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [20, 24, 28]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [12, 21]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [15, 18, 22, 23]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [12, 23]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [21, 27]}\n    ],\n    \"complexes\": [\"AF10-DOT1L complex\", \"SWI/SNF (proximity via GAS41/INI1)\"],\n    \"partners\": [\"DOT1L\", \"GAS41\", \"INI1/SMARCB1\", \"JAK1\", \"TCF4\", \"CTNNB1\", \"KAT5/Tip60\", \"CRM1/XPO1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}