ENAapplication/xmlftp.sra.ebi.ac.uk/vol1/fastq/SRR957/SRR957356/SRR957356.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR957/SRR957354/SRR957354.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR957/SRR957351/SRR957351.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR957/SRR957355/SRR957355.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR957/SRR957357/SRR957357.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR957/SRR957350/SRR957350.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR957/SRR957353/SRR957353.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR957/SRR957352/SRR957352.fastq.gzprimaryOK2000000GenomicsNIDDK, NIHhttps://www.ebi.ac.uk/ena/browser/view/PRJNA217217Mus musculusEnhancers play a central role in cell-type-specific gene expression and are marked by H3K4me1/2. Active enhancers are further marked by H3K27ac. However, the methyltransferases responsible for the deposition of H3K4me1/2 on enhancers remain elusive. Furthermore, the functions of these methyltransferases on enhancers and associated cell-type-specific gene expression are poorly understood. Here, we identify MLL4 (KMT2D) as a major H3K4 mono- and di-methyltransferase in mammalian cells. Using adipogenesis and myogenesis as model systems, we show that MLL4 exhibits cell-type- and differentiation-stage-specific genomic binding and is predominantly localized on enhancers. MLL4 co-localizes with lineage-determining transcription factors (TFs) on active enhancers during differentiation. Deletion of MLL4 dramatically decreases H3K4me1/2 and H3K27ac on enhancers and leads to severe defects in cell-type-specific gene expression and cell differentiation. Finally, we provide evidence that lineage-determining TFs recruit and require MLL4 to establish enhancers critical for cell-type-specific gene expression. Together, these results identify MLL4 as an H3K4 mono-/di-methyltransferase required for enhancer activation during cell differentiation. Overall design: ChIP-Seq analyses of MLL4 binding profiles at D0 (day 0), D2 (day 2), and D7 (day 7) of adipogenesis in WT (MLL3-/-) and MLL4 KO (MLL3-/-;MLL4-/-) brown preadipocytes. D0 and D2 input DNA samples can be found in GSE50417.ENABC032281, KABUK1, MLL2, Mll2, TNRC21, MLL4, C430014K11Rik, Mll4, wide, HRX2, WBP-7, ALR, KMT2B, wide/broad, CAGL114, Genomes, KMS, TRX2., AAD10, Maps, BC058659, broad, WBP7, MLL1Bmouse, mouse <Mus musculus>, house mouse.0.00.00.00.00.00falseMus musculusGenome-wide maps of MLL4 during brown preadipocyte differentiation2022-05-122013-12-19PRJNA217217GSE502092436873410090