| 2007 |
KMT2C/MLL3 is a catalytic subunit of a Set1-like histone H3K4 methyltransferase complex (PTIP complex/ASCOM) that also contains MLL4, ASH2L, RBBP5, WDR5, hDPY-30, NCOA6, PTIP, PA1, and UTX; the complex carries robust H3K4 methyltransferase activity. hDPY-30 binds ASH2L directly, and PA1 binds PTIP directly, requiring PTIP for interaction with the rest of the complex. |
Co-immunoprecipitation of endogenous complex, in vitro HMT activity assay, direct binding assays |
The Journal of biological chemistry |
High |
17500065
|
| 2009 |
KMT2C/MLL3 (within ASCOM complex) acts as a coactivator for p53 through direct interaction between p53 and ASCOM; ASCOM is required for H3K4 trimethylation and expression of endogenous p53 target genes in response to DNA damage (doxorubicin). Targeted inactivation of MLL3 H3K4-methylation activity in mice causes ureter epithelial tumors, exacerbated in p53+/- background. |
Co-IP demonstrating direct p53-ASCOM interaction; ChIP for H3K4me3 at p53 target genes; siRNA knockdown; mouse knock-in of catalytically inactive MLL3 |
Proceedings of the National Academy of Sciences of the United States of America |
High |
19433796
|
| 2008 |
KMT2C/MLL3 H3K4 methyltransferase activity within ASCOM is required for PPARγ-dependent adipogenesis; ASC-2, MLL3, and MLL4 are recruited to the PPARγ-activated aP2 gene during adipogenesis; PPARγ directly interacts with purified ASCOM. H3K4 methylation of aP2 is induced in WT but not in ASC-2-null MEFs and only partially induced in MLL3(Δ/Δ) MEFs. |
Mouse knock-in of catalytically inactive MLL3 (MLL3Δ/Δ), MEF adipogenesis assays, ChIP for H3K4 methylation, GST pulldown of PPARγ with ASCOM |
Proceedings of the National Academy of Sciences of the United States of America |
High |
19047629
|
| 2008 |
ASC-2 is required for ligand-induced recruitment of MLL3 and MLL4 to liver X receptors (LXRs); LXR ligand T1317 induces H3K4 trimethylation of LXR target genes in an ASC-2- and MLL3/4-dependent manner. ASC-2 functions as an essential adaptor to recruit MLL3/4 to LXRs. |
ChIP for H3K4me3 and complex components at LXR target gene promoters; ASC-2 null cells; MLL3(Δ/Δ) catalytically inactive mutant cells |
Molecular endocrinology (Baltimore, Md.) |
High |
18372346
|
| 2009 |
The C-terminal SET domain of MLL3 (and MLL4) directly interacts with INI1, an integral subunit of the SWI/SNF chromatin-remodeling complex; this interaction is required for mutual facilitation of ASCOM and SWI/SNF recruitment to nuclear receptor target genes. ASCOM and SWI/SNF are tightly colocalized in the nucleus. |
Co-IP of endogenous complexes, direct binding/pulldown assays, mutational analysis of SET domain, ChIP for complex recruitment to NR target genes |
Molecular endocrinology (Baltimore, Md.) |
High |
19221051
|
| 2011 |
MLL3 (as part of ASCOM) is required for H3K4me3-mediated activation of bile acid transporter genes BSEP, MRP2, and NTCP by FXR and glucocorticoid receptor; MLL3 and NCOA6 are recruited to FXR/GR elements at transporter promoters in a ligand-dependent manner; this recruitment and H3K4me3 is significantly diminished in mouse livers after bile duct ligation. |
ChIP in HepG2 cells and mouse liver, siRNA knockdown, GST pulldown of NCOA6 with FXR, luciferase reporter assay |
American journal of physiology. Gastrointestinal and liver physiology |
High |
21330447
|
| 2011 |
MLL3 complex is required for p53-dependent transactivation of SHP (small heterodimer partner) to regulate bile acid homeostasis; p53 directs recruitment of MLL3 complex components to p53-response elements of SHP; p53-dependent H3K4 trimethylation of SHP requires MLL3. |
ChIP in HepG2 cells and mouse liver (MLL3Δ/Δ mice), in vivo bile acid measurements |
Molecular endocrinology (Baltimore, Md.) |
High |
22034226
|
| 2013 |
KMT2C/MLL3 is a clock-controlled factor that directly and indirectly modulates circadian output gene expression in the liver; catalytic inactivation of MLL3 H3K4 methyltransferase activity severely compromises oscillation of core clock gene promoters (Bmal1, mCry1, mPer2, Rev-erbα), implicating rhythmic H3K4 methylation in circadian transcription. |
MLL3 catalytic-inactivation mouse model (MLL3Δ/Δ), ChIP-seq for H3K4 methylation at circadian gene loci, transcriptomic profiling |
Proceedings of the National Academy of Sciences of the United States of America |
High |
23297224
|
| 2013 |
MLL3 and MLL4 are major regulators of H3K4 monomethylation (H3K4me1) at enhancers in human cancer cells and mouse embryonic fibroblasts; ChIP-seq shows MLL4 is preferentially found at enhancer regions; loss of both MLL3 and MLL4 (in MLL3-mutant HCT116 cells with MLL4 knocked out) causes the greatest loss of H3K4me1 at enhancer regions. |
ChIP-seq for H3K4me1, H3K4me3, and MLL4 in HCT116 cells and MEFs; MLL4 knockout by homologous recombination |
Molecular and cellular biology |
High |
24081332
|
| 2013 |
In Drosophila, the MLL3/4 ortholog Trr restricts tissue growth and is required for H3K4 monomethylation; trr mutant clones display markedly reduced H3K4me1 without changes in H3K4me2/3; Trr and UTX (H3K27 demethylase) are found in the same protein complex, and trr mutant phenotype resembles Utx mutant phenotype. |
Drosophila genetic clonal analysis, immunostaining for H3K4me1/2/3 in trr mutant clones, co-complex analysis |
Molecular and cellular biology |
High |
23459941
|
| 2014 |
An ~50% reduction in MLL3 gene dosage (haploinsufficiency) impairs differentiation of hematopoietic stem and progenitor cells (HSPC) and cooperates with other -7/del(7q) events to promote leukemogenesis; established using RNAi and CRISPR/Cas9 approaches in mouse models. |
RNAi knockdown and CRISPR/Cas9 gene editing in mouse hematopoietic cells, transplantation assays, differentiation assays |
Cancer cell |
High |
24794707
|
| 2015 |
Somatic cancer mutations in the MLL3 SET domain alter catalytic properties: N4848S renders the enzyme inactive by disrupting cofactor binding; Y4884C converts MLL3 from a monomethyltransferase to a trimethyltransferase with H3K4me1 as preferred substrate; expression of Y4884C leads to aberrant H3K4me3 in cells. |
In vitro methyltransferase assays with purified mutant proteins, cellular expression of cancer mutants with histone modification analysis |
Clinical epigenetics |
High |
25829971
|
| 2015 |
WDR5 has a unique inhibitory role in the MLL3 core complex: unlike MLL1, MLL3 SET domain assembles with RbBP5/Ash2L independently of the Win motif-WDR5 interaction; WDR5 inhibits monomethylation activity of MLL3 core complex in a Win-motif-dependent manner; WRAD subcomplex catalyzes weak H3K4 monomethylation within the MLL3 core complex. |
In vitro reconstitution of MLL3 core complex, methyltransferase activity assays, small angle X-ray scattering (SAXS) for solution structure |
The Journal of biological chemistry |
High |
26324722
|
| 2015 |
MLL3 and MLL4 complexes bind directly to MAFA and MAFB transcription factors in islet β-cells; MLL3/4 and their complexes (~1.5 MDa) co-fractionate with MafA in size-fractionated β-cell extracts; knockdown of NCOA6 (core MLL3/4 subunit) reduces expression of a subset of MAFA/MAFB target genes. |
In-cell biochemical pulldown with mass spectrometry, size-exclusion fractionation, NCOA6 knockdown, gene expression analysis |
Diabetes |
Medium |
26180087
|
| 2017 |
Mll3/4 proteins (not their H3K4me1 catalytic activity) are required for enhancer RNA synthesis and Pol II occupancy at enhancers; loss of Mll3/4 proteins depletes enhancer Pol II and eRNA synthesis with downregulation of target genes exhibiting reduced polymerase in gene bodies (suggestive of pause-release defects). Loss of H3K4me1 alone (in catalytically deficient cells) has only minor effects on transcription. |
CRISPR/Cas9 genome editing to generate catalytically deficient vs protein-null Mll3/4 cells; ChIP-seq for H3K4me1, H3K27ac, Pol II; GRO-seq for nascent transcription |
Molecular cell |
High |
28483418
|
| 2017 |
MLL3/MLL4 are required for CBP/p300 binding on enhancers during adipogenesis; MLL3/MLL4 are required for super-enhancer formation; MLL4 and CBP identify super-enhancers of adipogenesis. The data suggest sequential enhancer priming by MLL3/MLL4 (H3K4me1/2) followed by enhancer activation by CBP/p300 (H3K27ac). |
ChIP-seq for MLL4, CBP, H3K4me1/2/3, H3K27ac, lineage TFs, Pol II; RNA-seq; ATAC-seq; conditional MLL3/4 knockout in brown adipocyte differentiation |
Nucleic acids research |
High |
28398509
|
| 2019 |
Extended PHD6 domain (ePHD6) of MLL3 and MLL4, involving the sixth PHD domain and its preceding zinc finger, specifically recognizes an H4H18-containing histone H4 fragment; modifications of residues surrounding H4H18 modulate H4 binding to MLL3/4. The interaction between ePHD6 and histone H4 is required for nucleosomal methylation activity and MLL4-mediated neuronal differentiation. |
Crystal structure of ePHD6-H4 complex, in vitro methyltransferase assays with nucleosomes, mutagenesis, cellular differentiation assays |
Nature communications |
High |
30604749
|
| 2019 |
H3.3K4M mutation prevents enhancer activation in adipogenesis specifically by destabilizing MLL3/4 proteins (but not other Set1-like H3K4 methyltransferases MLL1, MLL2, SET1A, SET1B); deletion of the enzymatic SET domain similarly destabilizes MLL3/4 proteins and impairs adipose tissue and muscle development. |
Transgenic mouse lines expressing H3.3K4M and conditional SET domain deletion; western blot for protein stability; ChIP-seq for enhancer marks; adipogenesis assays |
Nucleic acids research |
High |
30335158
|
| 2019 |
KMT2C/MLL3 knockdown in bladder cancer cells leads to deficiency in homologous recombination-mediated double-strand break DNA repair; cells with low KMT2C activity exhibit higher endogenous DNA damage and genomic instability, and rely on PARP1/2 for DNA repair, creating synthetic lethality with PARP inhibitor olaparib. |
KMT2C knockdown in bladder cancer cell lines, HR-repair assays, γH2AX/DNA damage quantification, PARP inhibitor sensitivity assays, ChIP for epigenetic marks at DNA repair genes |
EMBO reports |
Medium |
30665945
|
| 2018 |
KMT2C is necessary for hormone-driven ERα activity and estrogen-dependent breast cancer proliferation; KMT2C knockdown causes H3K4me1 and H3K27ac loss selectively at ERα enhancers; KMT2C loss suppresses estrogen-dependent gene expression but promotes tumor outgrowth under hormone-depleted conditions. |
KMT2C knockdown, ChIP-seq for H3K4me1 and H3K27ac at ERα enhancers, RNA-seq, estrogen-dependent proliferation assays |
Oncogene |
High |
29755131
|
| 2020 |
A small UTX stabilization domain (USD, ~80 amino acids) within Trr/MLL3/MLL4 binds and stabilizes UTX/KDM6A in vivo; nuclear UTX stability is enhanced when the USD is fused with the MLL4 HMG-box. COMPASS-dependent UTX stabilization is an essential non-catalytic function of Trr/MLL3/MLL4. |
Drosophila genetic rescue screen for minimal Trr domain, co-IP and stability assays for UTX, domain mapping with MLL3/4 human sequences |
Genes & development |
High |
33033055
|
| 2020 |
Cancer-derived UTX TPR mutations G137V and D336G impair UTX interaction with ASH2L and MLL3/4-specific components PTIP and PA1; interaction-compromised UTX mutants are preferentially localized to the cytoplasm, demonstrating that MLL3/4 complexes retain UTX in the nucleus through TPR-mediated interaction. WT UTX suppresses colony formation in soft agar while G137V fails, linking UTX-MLL3/4 interaction to tumor suppressor function. |
Co-IP of UTX mutants with MLL3/4 complex components, subcellular fractionation, CRISPR-Cas9 knock-in of G137V in HCT116, soft-agar colony formation assay |
Oncogene |
High |
32071397
|
| 2021 |
KMT2C/MLL3 is directly recruited to DNA damage sites by Ago2 and small noncoding DNA damage response RNA (DDR RNA); at damage sites, KMT2C mediates H3K4 methylation, chromatin relaxation, secondary recruitment of DDR factors, and amplification of DDR signals along chromatin. This is a transcription-independent role of KMT2C in DDR. |
ChIP at DNA damage sites, Ago2 co-IP with KMT2C, chromatin relaxation assays, HR repair assays, PARP inhibitor sensitivity assays in KMT2C/D-mutant cells |
Cancer research |
Medium |
33853832
|
| 2021 |
ASXL2 subunit of the BAP1 complex mediates a direct interaction with MLL3/COMPASS; ASXL2 loss results in decreased MLL3 occupancy at enhancers and reduced BAP1-MLL3 target gene expression. CARM1/PRMT4 methylates ASXL2 at R639/R641, which blocks ASXL2 binding to MLL3 and impairs expression of MLL3/COMPASS-dependent genes. |
Co-IP demonstrating ASXL2-MLL3 interaction, ChIP-seq for MLL3 occupancy upon ASXL2 loss, in vitro methylation of ASXL2 by CARM1, binding assays with methylated vs unmethylated ASXL2 |
Science advances |
High |
36197977
|
| 2021 |
Kmt2c deletions markedly enhance murine HSC self-renewal without altering proliferation rates; Kmt2c-deficient HSCs fail to differentiate appropriately in response to interleukin-1 when driven into cycle; Kmt2c deletions mitigate histone methylation/acetylation changes that normally accrue as HSCs cycle and impair enhancer recruitment during HSC differentiation. |
Mouse HSC transplantation assays, competitive repopulation assays, ChIP for histone marks, cytokine stimulation assays, RNA-seq |
Cell reports |
High |
33596429
|
| 2021 |
MLL3 binds the enhancer of PD-L1 and promotes its transcription; depletion of MLL3 decreases H3K4me1 at the PD-L1 enhancer and RNA Pol II Ser-5p at the PD-L1 promoter. |
ChIP for H3K4me1 and Pol II at PD-L1 locus, MLL3 depletion in prostate cancer cells, xenograft mouse experiments |
Biochimica et biophysica acta. Molecular basis of disease |
Medium |
30385408
|
| 2021 |
MLL3 depletion downregulates H3K4me1 and H3K27ac on an enhancer ~7 kb upstream of TNS3, reduces enhancer-promoter looping (by 3C assay), and impairs TNS3 expression; loss of TNS3 mediates the enhanced cell migration phenotype caused by MLL3 loss in cancer cells. |
CRISPR/sgRNA MLL3 depletion, RNA-seq, ChIP-seq for H3K4me1 and H3K27ac, 3C assay, dCas9-KRAB enhancer repression, TNS3 rescue experiments |
Cell death & disease |
Medium |
33824309
|
| 2021 |
MLL3 directly regulates DNMT3A expression through histone methylation at the DNMT3A locus; KMT2C deficiency leads to both histone H3K4 and DNA hypomethylation in SCLC; forced DNMT3A expression restrained metastasis of KMT2C-deficient SCLC through repressing MEIS/HOX genes. |
SCLC organoid-based mouse models, ChIP for H3K4 methylation at DNMT3A locus, WGBS for DNA methylation, RNA-seq, DNMT3A rescue experiments |
Nature cancer |
High |
35449309
|
| 2021 |
The MLL3 core complex catalyzes H3K4 methylation via a random sequential bi-bi kinetic mechanism (distinct from MLL4's ordered sequential bi-bi mechanism); for MLL3, AdoMet binding is NOT a prerequisite for H3 peptide binding; MLL4 requires AdoMet binding prior to H3 binding due to conformational stabilization of the active site. |
Steady-state kinetic analyses of MLL3 and MLL4 SET domain ternary complexes, inhibition studies, fluorescence polarization binding assays, molecular dynamics simulations |
The Journal of biological chemistry |
High |
33823156
|
| 2023 |
KMT2C/MLL3 co-activates the CDKN2A tumor suppressor locus by binding to it and establishing H3K4 methylation; disruption of Kmt2c cooperates with Myc overexpression in hepatocellular carcinoma with blunted MLL3 binding at Cdkn2a, reduced H3K4 methylation, and low p16/Ink4a and p19/Arf expression. Endogenous Kmt2c restoration reverses chromatin/transcriptional effects and triggers Ink4a/Arf-dependent apoptosis. |
ChIP-seq for MLL3 and H3K4 methylation at Cdkn2a, CRISPR and conditional Kmt2c knockout mouse model, liver organoid cancer models, genetic rescue experiments |
eLife |
High |
37261974
|
| 2023 |
MLL3 loss leads to increased IFNγ signaling in breast cancer cells, which contributes to hybrid EMT induction and enhanced metastatic capacity; MLL3 loss promotes phenotypic plasticity (hybrid EMT) rather than unidirectional EMT, and BET inhibition suppresses MLL3-mutant tumor growth. |
MLL3 deletion in breast cancer cells, transcriptomic profiling, IFNγ signaling inhibition experiments, in vivo metastasis assays, BET inhibitor treatment |
Nature cell biology |
Medium |
36604594
|
| 2023 |
Constitutive elimination of both MLL3 and MLL4 enzymatic activities prevents gastrulation and causes early embryonic lethality; selective elimination of MLL3/4 enzymatic activities in embryonic (but not extraembryonic) lineages leaves gastrulation largely intact. MLL3/4-catalyzed H3K4me1 is largely dispensable for enhancer activation during ESC differentiation but required for GATA6 binding at ExEn enhancers. |
Conditional and constitutive MLL3/4 SET-domain knockout mouse models, ChIP-seq for H3K4me1, RNA-seq, embryo phenotyping |
Nature genetics |
High |
37012455
|
| 2023 |
Loss of MLL3/4 in ESCs uncouples H3K4me1, H3K27ac, and gene activation: MLL3/4 activity is required at most sites that dynamically gain or lose H3K4me1 but dispensable at stably methylated sites; many sites gain H3K27ac independent of MLL3/4 or H3K4me1; transcriptional activation of nearby genes is largely unaffected despite failure to gain active histone marks at thousands of enhancers. |
MLL3/4 double knockout ESCs, ChIP-seq for H3K4me1, H3K27ac, and Pol II, RNA-seq during early ESC differentiation |
Genome biology |
High |
36869380
|
| 2023 |
Loss of Kmt2c in TNBC drives brain metastasis through KDM6A-dependent upregulation of MMP3: Kmt2c knockout alters H3K4me1, H3K27ac, and H3K27me3 chromatin marks and enhances KDM6A binding at loci correlating with gene expression; KDM6A downregulation or inhibition diminishes Mmp3 upregulation and prevents brain metastasis similarly to direct Mmp3 knockdown. |
TNBC mouse models with Kmt2c or Kmt2d deletion, ChIP-seq for multiple histone marks, RNA-seq, KDM6A inhibition, in vivo metastasis assays |
Nature cell biology |
High |
38926506
|
| 2024 |
KMT2C binds to active enhancers and CpG-poor promoters regulating urothelial lineage programs; Kmt2c/d knockout leads to diminished H3K4me1, H3K27ac, and nascent RNA transcription at these sites causing impaired urothelial differentiation; Kmt2c/d loss causes redistribution of KMT2A-menin from KMT2D-occupied enhancers to CpG-high and bivalent promoters, derepressing signal-induced immediate early genes. |
Genetically engineered mouse models with urothelium-specific Kmt2c/d knockout, ChIP-seq for H3K4me1, H3K27ac, KMT2D, KMT2A-menin, nascent RNA-seq, EGFR inhibitor sensitivity assays |
Nature genetics |
High |
39806204
|
| 2025 |
KMT2C maintains ASPP2 expression via enhancer-promoter communication in response to AR inhibition; KMT2C inactivation reduces ASPP2, triggering ΔNp63-dependent luminal-to-DNPC transdifferentiation in prostate cancer; the DNPC state maintains fatty acid synthesis through ΔNp63-mediated SREBP1c transactivation, fueling DNPC growth via HRAS palmitoylation and MAPK signaling. |
KMT2C conditional knockout in prostate organoids and mouse models, ChIP-seq for enhancer-promoter interactions, ChIA-PET/Hi-ChIP, RNA-seq, rescue experiments with ASPP2, lipid metabolism assays |
Cancer cell |
Medium |
40280125
|
| 2025 |
MLL3 loss stabilizes HIF1α in breast cancer cells, leading to increased CCL2 secretion by tumor cells and recruitment of CCR2+ regulatory T cells (Tregs); tumor-infiltrating Tregs differentiate into ICOShiGITRhi effectors secreting TGF-β and IL-10 in a BLIMP-1-dependent manner; antibody targeting of ICOS or GITR depletes tumor Tregs and inhibits tumorigenesis. |
Mouse mammary-stem-cell-based tumor model with MLL3/p53 loss and PI3K activation, HIF1α stabilization assays, CCL2 secretion ELISA, Treg depletion experiments, anti-ICOS/GITR antibody treatment, transcriptomic profiling of Tregs |
Immunity |
Medium |
40749665
|
| 2016 |
A PTIP-PA1 subcomplex functions independently of MLL3/MLL4 in promoting transcription for immunoglobulin class switch recombination; quantitative proteomics in primary lymphocytes identified a tandem BRCT domain of PTIP sufficient for CSR, with PA1 as its main functional partner, functioning separately from the MLL3/MLL4 complex and from PTIP's DNA damage localization. |
Quantitative proteomics in primary lymphocytes, domain mapping of PTIP, CSR assays in B cells, domain-deletion mutant analysis |
Genes & development |
Medium |
26744420
|
| 2012 |
Drosophila cara mitad (cmi), encoding the N-terminal PHD/HMG portion of the ancestral MLL2/3 gene, associates with TRR (the C-terminal SET domain protein, ortholog of MLL3/4) and the EcR-USP nuclear receptor; CMI is required for hormone-dependent transcription. Genetic tests show that hormone-stimulated transcription requires CMI chromatin binding, H3K4 methylation by TRR, and H3K27 demethylation by UTX. |
Co-IP of CMI with TRR and EcR-USP, genetic epistasis in Drosophila, ChIP for H3K4me3 in cmi mutants |
Development (Cambridge, England) |
Medium |
22569554
|
| 2022 |
KMT2C methyltransferase activity regulates p16INK4A expression; impaired KMT2C methyltransferase activity (SET domain deletion) in mouse prostate drives proliferation, PIN formation, and combined with PTEN loss triggers senescence loss and metastasis. KMT2C-mutated tumors show enrichment of MYC gene signatures and loss of p16INK4A expression. |
Conditional Kmt2c SET domain deletion in mouse prostate epithelium, Pten-deficient mouse model, ChIP for H3K4me at INK4A locus, RNA-seq, in vivo tumor phenotyping |
Molecular cancer |
Medium |
35354467
|
| 2022 |
YTHDC1 binds to and stabilizes m6A-modified KMT2C mRNA; YTHDC1 knockdown reduces KMT2C protein levels and compromises DDR via reduced H3K4 methylation at DDR gene loci in B-ALL cells. |
RNA immunoprecipitation (RIP) for YTHDC1-KMT2C mRNA interaction, m6A-seq, KMT2C protein stability assays, ChIP for H3K4 methylation at DDR genes, YTHDC1 knockdown with phenotypic rescue by KMT2C |
Leukemia |
Medium |
39501105
|
| 2020 |
MLL3/MLL4-associated PAGR1 (PA1) cooperates with phosphorylated CREB and ligand-activated glucocorticoid receptor to directly control induction of C/EBPβ and C/EBPδ in early adipogenesis; deletion of Pagr1 prevents induction of C/EBPβ/δ and severely impairs adipogenesis, rescued by ectopic C/EBPβ or PPARγ expression. |
Conditional Pagr1 deletion in white/brown preadipocytes (Myf5+ precursors), adipogenesis assays, ChIP for PAGR1 and CREB at C/EBPβ/δ loci, rescue with C/EBPβ/PPARγ expression |
Molecular and cellular biology |
Medium |
32601106
|