Abstract
Histone H3 Lys4 (H3K4) methylation is a prevalent mark associated with transcription activation. A common feature of several H3K4 methyltransferase complexes is the presence of three structural components (RbBP5, Ash2L and WDR5) and a catalytic subunit containing a SET domain. Here we report the first biochemical reconstitution of a functional four-component mixed-lineage leukemia protein-1 (MLL1) core complex. This reconstitution, combined with in vivo assays, allows direct analysis of the contribution of each component to MLL1 enzymatic activity and their roles in transcriptional regulation. Moreover, taking clues from a crystal structure analysis, we demonstrate that WDR5 mediates interactions of the MLL1 catalytic unit both with the common structural platform and with the histone substrate. Mechanistic insights gained from this study can be generalized to the whole family of SET1-like histone methyltransferases in mammals.
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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Catalytic Domain
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Cell Line
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism
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Heterotrimeric GTP-Binding Proteins / chemistry
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Heterotrimeric GTP-Binding Proteins / metabolism*
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Histone-Lysine N-Methyltransferase / genetics
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Histone-Lysine N-Methyltransferase / metabolism*
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Histones / metabolism
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Humans
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Intracellular Signaling Peptides and Proteins
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Lysine / metabolism*
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Methylation
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Multiprotein Complexes
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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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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism
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RNA Interference
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Recombinant Proteins / chemistry
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Recombinant Proteins / metabolism
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Transcription Factors / genetics
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Transcription Factors / metabolism
Substances
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ASH2L protein, human
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DNA-Binding Proteins
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Histones
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Intracellular Signaling Peptides and Proteins
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KMT2A protein, human
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Multiprotein Complexes
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Nuclear Proteins
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RBBP5 protein, human
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Recombinant Proteins
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Transcription Factors
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WDR5 protein, human
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Myeloid-Lymphoid Leukemia Protein
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Histone-Lysine N-Methyltransferase
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Heterotrimeric GTP-Binding Proteins
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Lysine