Binding of the MLL PHD3 finger to histone H3K4me3 is required for MLL-dependent gene transcription.
The MLL (mixed-lineage leukemia) proto-oncogene encodes a histone methyltransferase that creates the methylated histone H3K4 epigenetic marks, commonly associated with actively transcribed genes. In addition to its canonical histone methyltransferase SET domain, the MLL protein contains three plant homeodomain (PHD) fingers that are well conserved between species but whose potential roles and requirements for MLL function are unknown. Here, we demonstrate that the third PHD domain of MLL (PHD3) binds histone H3 trimethylated at lysine 4 (H3K4me3) with high affinity and specificity and H3K4me2 with 8-fold lower affinity. Biochemical and structural analyses using NMR and fluorescence spectroscopy identified key amino acids essential for the interaction with H3K4me3. Site-directed mutations of the residues involved in recognition of H3K4me3 compromised in vitro H3K4me3 binding but not in vivo localization of full-length MLL to chromatin sites in target promoters of MEIS1 and HOXA genes. Whereas intact PHD3 finger was necessary for MLL occupancy at these promoters, H3K4me3 binding was critical for MLL transcriptional activity. These results demonstrate that MLL occupancy and target gene activation can be functionally separated. Furthermore, these findings reveal that MLL not only "writes" the H3K4me3 mark but also binds the mark, and this binding is required for the transcriptional maintenance functions of MLL.
Chang
Pei-Yun
PY
Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Hom
Robert A
RA
Musselman
Catherine A
CA
Zhu
Li
L
Kuo
Alex
A
Gozani
Or
O
Kutateladze
Tatiana G
TG
Cleary
Michael L
ML
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002460:Cell Line; D005786:Gene Expression Regulation; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008239:Lysine / Q000235:genetics / Q000378:metabolism*; D008745:Methylation; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D051788:Myeloid-Lymphoid Leukemia Protein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D000090063:Proto-Oncogene Mas; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D014158:Transcription, Genetic
D006657:Histones; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D011993:Recombinant Fusion Proteins; D051788:Myeloid-Lymphoid Leukemia Protein; D008239:Lysine
Proteins. 2005 Jun 1;59(4):687-96
15815974
Biochemistry. 1989 Jul 25;28(15):6150-6
2675964
Biochemistry. 1997 Apr 1;36(13):3959-70
9092826
Mol Cell Biol. 2001 May;21(10):3589-97
11313484
Proc Natl Acad Sci U S A. 2005 Oct 11;102(41):14481-2
16203969
Hematol Oncol. 2005 Mar;23(1):1-9
16118769
Mutat Res. 2008 Dec 1;647(1-2):3-12
18682256
Nature. 2006 Jul 6;442(7098):91-5
16728978
Biochemistry. 1996 Aug 13;35(32):10256-61
8756680
J Biol Chem. 2007 Jan 26;282(4):2450-5
17142463
Nature. 2006 Jul 6;442(7098):86-90
16728976
Hum Mol Genet. 2001 Sep 1;10(18):1995-2011
11555636
Nature. 2006 Jul 6;442(7098):96-9
16728974
Nature. 2006 Jul 6;442(7098):100-3
16728977
J Biomol NMR. 1995 Nov;6(3):277-93
8520220
Mol Interv. 2009 Dec;9(6):314-23
20048137
Biochemistry. 1989 Mar 21;28(6):2387-91
2730871