title: DOT1L/KMT4 recruitment and H3K79 methylation are ubiquitously coupled with gene transcription in mammalian cells.
abstract: The histone H3 lysine 79 methyltransferase DOT1L/KMT4 can promote an oncogenic pattern of gene expression through binding with several MLL fusion partners found in acute leukemia. However, the normal function of DOT1L in mammalian gene regulation is poorly understood. Here we report that DOT1L recruitment is ubiquitously coupled with active transcription in diverse mammalian cell types. DOT1L preferentially occupies the proximal transcribed region of active genes, correlating with enrichment of H3K79 di- and trimethylation. Furthermore, Dot1l mutant fibroblasts lacked H3K79 di- and trimethylation at all sites examined, indicating that DOT1L is the sole enzyme responsible for these marks. Importantly, we identified chromatin immunoprecipitation (ChIP) assay conditions necessary for reliable H3K79 methylation detection. ChIP-chip tiling arrays revealed that levels of all degrees of genic H3K79 methylation correlate with mRNA abundance and dynamically respond to changes in gene activity. Conversion of H3K79 monomethylation into di- and trimethylation correlated with the transition from low- to high-level gene transcription. We also observed enrichment of H3K79 monomethylation at intergenic regions occupied by DNA-binding transcriptional activators. Our findings highlight several similarities between the patterning of H3K4 methylation and that of H3K79 methylation in mammalian chromatin, suggesting a widespread mechanism for parallel or sequential recruitment of DOT1L and MLL to genes in their normal "on" state.
authors.affiliation: Division of Hematology, Abramson Research Center 315A, The Children's Hospital of Philadelphia, 3400 Civic Center Blvd., Philadelphia, PA 19104, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Steger
authors.forename: Martina I
authors.initials: MI
authors.lastname: Lefterova
authors.forename: Lei
authors.initials: L
authors.lastname: Ying
authors.forename: Aaron J
authors.initials: AJ
authors.lastname: Stonestrom
authors.forename: Michael
authors.initials: M
authors.lastname: Schupp
authors.forename: David
authors.initials: D
authors.lastname: Zhuo
authors.forename: Adam L
authors.initials: AL
authors.lastname: Vakoc
authors.forename: Ja-Eun
authors.initials: JE
authors.lastname: Kim
authors.forename: Junjie
authors.initials: J
authors.lastname: Chen
authors.forename: Mitchell A
authors.initials: MA
authors.lastname: Lazar
authors.forename: Gerd A
authors.initials: GA
authors.lastname: Blobel
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Vakoc
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000378:metabolism; D000818:Animals; D018955:CD36 Antigens / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D002843:Chromatin / Q000235:genetics; D047369:Chromatin Immunoprecipitation; D050982:GATA1 Transcription Factor / Q000378:metabolism; D005914:Globins / Q000235:genetics / Q000378:metabolism; D011495:Histone-Lysine N-Methyltransferase; D006657:Histones / Q000378:metabolism*; D008239:Lysine / Q000235:genetics / Q000378:metabolism; D008745:Methylation; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D047495:PPAR gamma / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate; D014158:Transcription, Genetic / Q000235:genetics*
chemical_list: D018955:CD36 Antigens; D002843:Chromatin; D050982:GATA1 Transcription Factor; C494689:Gata1 protein, mouse; D006657:Histones; D047495:PPAR gamma; D012967:Sodium Dodecyl Sulfate; D005914:Globins; C473477:DOT1L protein, human; C481527:Dot1l protein, mouse; D008780:Methyltransferases; D011495:Histone-Lysine N-Methyltransferase; D008239:Lysine