title: Targeting recruitment of disruptor of telomeric silencing 1-like (DOT1L): characterizing the interactions between DOT1L and mixed lineage leukemia (MLL) fusion proteins.
abstract: The MLL fusion proteins, AF9 and ENL, activate target genes in part via recruitment of the histone methyltransferase DOT1L (disruptor of telomeric silencing 1-like). Here we report biochemical, biophysical, and functional characterization of the interaction between DOT1L and MLL fusion proteins, AF9/ENL. The AF9/ENL-binding site in human DOT1L was mapped, and the interaction site was identified to a 10-amino acid region (DOT1L865-874). This region is highly conserved in DOT1L from a variety of species. Alanine scanning mutagenesis analysis shows that four conserved hydrophobic residues from the identified binding motif are essential for the interactions with AF9/ENL. Binding studies demonstrate that the entire intact C-terminal domain of AF9/ENL is required for optimal interaction with DOT1L. Functional studies show that the mapped AF9/ENL interacting site is essential for immortalization by MLL-AF9, indicating that DOT1L interaction with MLL-AF9 and its recruitment are required for transformation by MLL-AF9. These results strongly suggest that disruption of interaction between DOT1L and AF9/ENL is a promising therapeutic strategy with potentially fewer adverse effects than enzymatic inhibition of DOT1L for MLL fusion protein-associated leukemia.
authors.affiliation: From the Department of Pathology and; the Chemical Biology Doctoral Program, University of Michigan Medical School, Ann Arbor, Michigan 48109 and.
authors.forename: Chenxi
authors.initials: C
authors.lastname: Shen
authors.affiliation: From the Department of Pathology and.
authors.forename: Stephanie Y
authors.initials: SY
authors.lastname: Jo
authors.affiliation: the School of Medical Engineering, Hefei University of Technology, Hefei, Anhui 230009, China.
authors.forename: Chenzhong
authors.initials: C
authors.lastname: Liao
authors.affiliation: From the Department of Pathology and; the Chemical Biology Doctoral Program, University of Michigan Medical School, Ann Arbor, Michigan 48109 and.
authors.forename: Jay L
authors.initials: JL
authors.lastname: Hess
authors.affiliation: From the Department of Pathology and; the Chemical Biology Doctoral Program, University of Michigan Medical School, Ann Arbor, Michigan 48109 and. Electronic address: zanetan@med.umich.edu.
authors.forename: Zaneta
authors.initials: Z
authors.lastname: Nikolovska-Coleska
mesh_terms: D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D057809:HEK293 Cells; D011495:Histone-Lysine N-Methyltransferase; D006801:Humans; D008780:Methyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D051788:Myeloid-Lymphoid Leukemia Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015514:Oncogene Proteins, Fusion / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D035602:Transcriptional Elongation Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: C090862:ELL protein, human; C497422:KMT2A protein, human; C489901:MLLT3 protein, human; D009687:Nuclear Proteins; D015514:Oncogene Proteins, Fusion; D035602:Transcriptional Elongation Factors; D051788:Myeloid-Lymphoid Leukemia Protein; C473477:DOT1L protein, human; D008780:Methyltransferases; D011495:Histone-Lysine N-Methyltransferase
keywords: AF9; DOT1L; ENL; Fusion Protein; Histone Methylation; Mixed Lineage Leukemia; Peptides; Protein-Protein Interactions; Surface Plasmon Resonance (SPR)