Abstract
The MLL gene is a common target of chromosomal translocations found in human leukemia. MLL-fusion leukemia has a consistently poor outcome. One of the most common translocation partners is AF9 (MLLT3). MLL-AF9 recruits DOT1L, a histone 3 lysine 79 methyltransferase (H3K79me1/me2/me3), leading to aberrant gene transcription. We show that DOT1L has three AF9 binding sites and present the nuclear magnetic resonance (NMR) solution structure of a DOT1L-AF9 complex. We generate structure-guided point mutations and find that they have graded effects on recruitment of DOT1L to MLL-AF9. Chromatin immunoprecipitation sequencing (ChIP-seq) analyses of H3K79me2 and H3K79me3 show that graded reduction of the DOT1L interaction with MLL-AF9 results in differential loss of H3K79me2 and me3 at MLL-AF9 target genes. Furthermore, the degree of DOT1L recruitment is linked to the level of MLL-AF9 hematopoietic transformation.
Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Binding Sites
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Chromatin Immunoprecipitation
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Histone-Lysine N-Methyltransferase
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Histones / metabolism*
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Humans
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Magnetic Resonance Spectroscopy
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Methylation
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Methyltransferases / chemistry
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Methyltransferases / genetics
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Methyltransferases / metabolism*
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Molecular Sequence Data
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Myeloid-Lymphoid Leukemia Protein / chemistry
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Myeloid-Lymphoid Leukemia Protein / genetics
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Myeloid-Lymphoid Leukemia Protein / metabolism*
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Oncogene Proteins, Fusion / chemistry
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Oncogene Proteins, Fusion / genetics
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Oncogene Proteins, Fusion / metabolism*
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Point Mutation
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Protein Binding
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Protein Structure, Tertiary
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Sequence Analysis, DNA
Substances
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Histones
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MLL-AF9 fusion protein, human
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Oncogene Proteins, Fusion
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Recombinant Proteins
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Myeloid-Lymphoid Leukemia Protein
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DOT1L protein, human
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Methyltransferases
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Histone-Lysine N-Methyltransferase