Physical Association and Coordinate Function of the H3 K4 Methyltransferase MLL1 and the H4 K16 Acetyltransferase MOF

A stable complex containing MLL1 and MOF has been immunoaffinity purified from a human cell line that stably expresses an epitope-tagged WDR5 subunit. Stable interactions between MLL1 and MOF were confirmed by reciprocal immunoprecipitation, cosedimentation, and cotransfection analyses, and interaction sites were mapped to MLL1 C-terminal and MOF zinc finger domains. The purified complex has a robust MLL1-mediated histone methyltransferase activity that can effect mono-, di-, and trimethylation of H3 K4 and a MOF-mediated histone acetyltransferase activity that is specific for H4 K16. Importantly, both activities are required for optimal transcription activation on a chromatin template in vitro and on an endogenous MLL1 target gene, Hox a9, in vivo. These results indicate an activator-based mechanism for joint MLL1 and MOF recruitment and targeted methylation and acetylation and provide a molecular explanation for the closely correlated distribution of H3 K4 methylation and H4 K16 acetylation on active genes.

[1]  Paul Tempst,et al.  Proteolytic Cleavage of MLL Generates a Complex of N- and C-Terminal Fragments That Confers Protein Stability and Subnuclear Localization , 2003, Molecular and Cellular Biology.

[2]  R. Roeder,et al.  Ordered Cooperative Functions of PRMT1, p300, and CARM1 in Transcriptional Activation by p53 , 2004, Cell.

[3]  Kevin Struhl,et al.  Targeted recruitment of Set1 histone methylase by elongating Pol II provides a localized mark and memory of recent transcriptional activity. , 2003, Molecular cell.

[4]  A. Burlingame,et al.  Histone Acetylation and Deacetylation , 2002, Molecular & Cellular Proteomics.

[5]  S. Korsmeyer,et al.  MLL and CREB Bind Cooperatively to the Nuclear Coactivator CREB-Binding Protein , 2001, Molecular and Cellular Biology.

[6]  P. Meltzer,et al.  Activating Signal Cointegrator 2 Belongs to a Novel Steady-State Complex That Contains a Subset of Trithorax Group Proteins , 2003, Molecular and Cellular Biology.

[7]  T. Richmond,et al.  Preparation of nucleosome core particle from recombinant histones. , 1999, Methods in enzymology.

[8]  Z. Wang,et al.  Activator-dependent transcription from chromatin in vitro involving targeted histone acetylation by p300. , 2000, Molecular cell.

[9]  J. Tamkun,et al.  Modulation of ISWI function by site‐specific histone acetylation , 2002, EMBO reports.

[10]  Christian Beisel,et al.  Histone methylation by the Drosophila epigenetic transcriptional regulator Ash1 , 2002, Nature.

[11]  R. Roeder,et al.  Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. , 1983, Nucleic acids research.

[12]  C. Allis,et al.  Linking Global Histone Acetylation to the Transcription Enhancement of X-chromosomal Genes in Drosophila Males* , 2001, The Journal of Biological Chemistry.

[13]  Tony Kouzarides,et al.  Histone H3 lysine 4 methylation patterns in higher eukaryotic genes , 2004, Nature Cell Biology.

[14]  G. Kay,et al.  Menin associates with a trithorax family histone methyltransferase complex and with the hoxc8 locus. , 2004, Molecular cell.

[15]  S. Korsmeyer,et al.  Altered Hox expression and segmental identity in Mll-mutant mice , 1995, Nature.

[16]  Danny Reinberg,et al.  The constantly changing face of chromatin. , 2003, Science of aging knowledge environment : SAGE KE.

[17]  B. Chait,et al.  Rapidly switchable matrix-assisted laser desorption/ionization and electrospray quadrupole-time-of-flight mass spectrometry for protein identification , 2000, Journal of the American Society for Mass Spectrometry.

[18]  Stuart L Schreiber,et al.  Methylation of histone H3 K4 mediates association of the Isw1p ATPase with chromatin. , 2003, Molecular cell.

[19]  John R. Yates,et al.  Chd1 chromodomain links histone H3 methylation with SAGA- and SLIK-dependent acetylation , 2005, Nature.

[20]  C. Allis,et al.  The language of covalent histone modifications , 2000, Nature.

[21]  J. Lucchesi,et al.  A new human member of the MYST family of histone acetyl transferases with high sequence similarity to Drosophila MOF. , 2000, Biochimica et biophysica acta.

[22]  J. Workman,et al.  Histone Acetyltransferase Complexes Stabilize SWI/SNF Binding to Promoter Nucleosomes , 2001, Cell.

[23]  Karl Mechtler,et al.  Methylation of histone H3 lysine 9 creates a binding site for HP1 proteins , 2001, Nature.

[24]  T. Straub,et al.  Functional integration of the histone acetyltransferase MOF into the dosage compensation complex , 2004, The EMBO journal.

[25]  Nevan J. Krogan,et al.  COMPASS, a Histone H3 (Lysine 4) Methyltransferase Required for Telomeric Silencing of Gene Expression* , 2002, The Journal of Biological Chemistry.

[26]  O. Rozenblatt-Rosen,et al.  Menin and MLL cooperatively regulate expression of cyclin-dependent kinase inhibitors. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[27]  Jacques Côté,et al.  The diverse functions of histone acetyltransferase complexes. , 2003, Trends in genetics : TIG.

[28]  R. Roeder,et al.  Reconstitution and transcriptional analysis of chromatin in vitro. , 2004, Methods in enzymology.

[29]  R. Roeder,et al.  Selective requirements for histone H3 and H4 N termini in p300-dependent transcriptional activation from chromatin. , 2002, Molecular cell.

[30]  P. Becker,et al.  Activation of transcription through histone H4 acetylation by MOF, an acetyltransferase essential for dosage compensation in Drosophila. , 2000, Molecular cell.

[31]  B. Turner,et al.  Cellular Memory and the Histone Code , 2002, Cell.

[32]  C. Allis,et al.  Methylation of histone H3 at lysine 4 is highly conserved and correlates with transcriptionally active nuclei in Tetrahymena. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[33]  J. Hess.  MLL: a histone methyltransferase disrupted in leukemia. , 2004, Trends in molecular medicine.

[34]  Thomas A. Milne,et al.  WDR5 Associates with Histone H3 Methylated at K4 and Is Essential for H3 K4 Methylation and Vertebrate Development , 2005, Cell.

[35]  Danny Reinberg,et al.  Histone lysine methylation: a signature for chromatin function. , 2003, Trends in genetics : TIG.

[36]  E. Bradbury,et al.  A mass spectrometric “Western blot” to evaluate the correlations between histone methylation and histone acetylation , 2004, Proteomics.

[37]  W. Herr,et al.  Leukemia Proto-Oncoprotein MLL Forms a SET1-Like Histone Methyltransferase Complex with Menin To Regulate Hox Gene Expression , 2004, Molecular and Cellular Biology.

[38]  Stuart L. Schreiber,et al.  Active genes are tri-methylated at K4 of histone H3 , 2002, Nature.

[39]  B. Morgan,et al.  Genetic analysis of histone H4: essential role of lysines subject to reversible acetylation. , 1990, Science.

[40]  M. Grunstein,et al.  Genetic evidence for an interaction between SIR3 and histone H4 in the repression of the silent mating loci in Saccharomyces cerevisiae. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[41]  B. Chait,et al.  Automatic identification of proteins with a MALDI-quadrupole ion trap mass spectrometer. , 2001, Analytical chemistry.

[42]  B. Turner,et al.  Histone H4 isoforms acetylated at specific lysine residues define individual chromosomes and chromatin domains in Drosophila polytene nuclei , 1992, Cell.

[43]  Philip R Gafken,et al.  Mass spectrometric quantification of acetylation at specific lysines within the amino-terminal tail of histone H4. , 2003, Analytical biochemistry.

[44]  K. Yamamura,et al.  Analysis of the murine Hoxa-9 cDNA: an alternatively spliced transcript encodes a truncated protein lacking the homeodomain. , 1998, Gene.

[45]  W. Herr,et al.  Human Sin3 deacetylase and trithorax-related Set1/Ash2 histone H3-K4 methyltransferase are tethered together selectively by the cell-proliferation factor HCF-1. , 2003, Genes & development.

[46]  C. Allis,et al.  Histone and chromatin cross-talk. , 2003, Current opinion in cell biology.

[47]  Toshiki Mori,et al.  ALL-1 is a histone methyltransferase that assembles a supercomplex of proteins involved in transcriptional regulation. , 2002, Molecular cell.

[48]  C. Allis,et al.  The Drosophila MSL Complex Acetylates Histone H4 at Lysine 16, a Chromatin Modification Linked to Dosage Compensation , 2000, Molecular and Cellular Biology.

[49]  J. Workman,et al.  Analysis of nucleosome disruption by ATP-driven chromatin remodeling complexes. , 1999, Methods in molecular biology.

[50]  C. Croce,et al.  Trithorax and dCBP Acting in a Complex to Maintain Expression of a Homeotic Gene , 2001, Science.

[51]  Steven J Altschuler,et al.  Genomic characterization reveals a simple histone H4 acetylation code. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[52]  Thomas A Milne,et al.  MLL targets SET domain methyltransferase activity to Hox gene promoters. , 2002, Molecular cell.