title: Validation and structural characterization of the LEDGF/p75-MLL interface as a new target for the treatment of MLL-dependent leukemia.
abstract: Mixed lineage leukemia (MLL) fusion-driven acute leukemias represent a genetically distinct subset of leukemias with poor prognosis. MLL forms a ternary complex with the lens epithelium-derived growth factor (LEDGF/p75) and MENIN. LEDGF/p75, a chromatin reader recognizing H3K36me3 marks, contributes to the association of the MLL multiprotein complex to chromatin. Formation of this complex is critical for the development of MLL leukemia. Available X-ray data represent only a partial structure of the LEDGF/p75-MLL-MENIN complex. Using nuclear magnetic resonance spectroscopy, we identified an additional LEDGF/p75-MLL interface, which overlaps with the binding site of known LEDGF/p75 interactors-HIV-1 integrase, PogZ, and JPO2. Binding of these proteins or MLL to LEDGF/p75 is mutually exclusive. The resolved structure, as well as mutational analysis, shows that the interaction is primarily sustained via two aromatic residues of MLL (F148 and F151). Colony-forming assays in MLL-AF9(+) leukemic cells expressing MLL interaction-defective LEDGF/p75 mutants revealed that this interaction is essential for transformation. Finally, we show that the clonogenic growth of primary murine MLL-AF9-expressing leukemic blasts is selectively impaired upon overexpression of a LEDGF/p75-binding cyclic peptide CP65, originally developed to inhibit the LEDGF/p75-HIV-1 integrase interaction. The newly defined protein-protein interface therefore represents a new target for the development of therapeutics against LEDGF/p75-dependent MLL fusion-driven leukemic disorders. Cancer Res; 74(18); 5139-51. ©2014 AACR.
authors.affiliation: KU Leuven, Laboratory for Molecular Virology and Gene Therapy, Department of Pharmaceutical and Pharmacological Sciences, Leuven, Belgium.
authors.forename: Kateřina
authors.initials: K
authors.lastname: Cermáková
authors.affiliation: Institute of Organic Chemistry and Biochemistry of the ASCR, v.v.i., Structural Biology, Prague, Czech Republic. Charles University in Prague, Department of Genetics and Microbiology, Faculty of Science, Prague, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Tesina
authors.affiliation: KU Leuven, Laboratory for Molecular Virology and Gene Therapy, Department of Pharmaceutical and Pharmacological Sciences, Leuven, Belgium.
authors.forename: Jonas
authors.initials: J
authors.lastname: Demeulemeester
authors.affiliation: KU Leuven, Laboratory for Molecular Virology and Gene Therapy, Department of Pharmaceutical and Pharmacological Sciences, Leuven, Belgium.
authors.forename: Sara
authors.initials: S
authors.lastname: El Ashkar
authors.affiliation: Department of Biomedicine, University Hospital and Children's Hospital Basel (UKBB) ZLF, Basel, Switzerland.
authors.forename: Hélène
authors.initials: H
authors.lastname: Méreau
authors.affiliation: Department of Biomedicine, University Hospital and Children's Hospital Basel (UKBB) ZLF, Basel, Switzerland.
authors.forename: Juerg
authors.initials: J
authors.lastname: Schwaller
authors.affiliation: Institute of Organic Chemistry and Biochemistry of the ASCR, v.v.i., Structural Biology, Prague, Czech Republic. Institute of Molecular Genetics of the ASCR, v.v.i., Structural Biology, Prague, Czech Republic.
authors.forename: Pavlína
authors.initials: P
authors.lastname: Rezáčová
authors.affiliation: Institute of Organic Chemistry and Biochemistry of the ASCR, v.v.i., Structural Biology, Prague, Czech Republic. Jan.Derijck@med.kuleuven.be veverka@uochb.cas.cz.
authors.forename: Vaclav
authors.initials: V
authors.lastname: Veverka
authors.affiliation: KU Leuven, Laboratory for Molecular Virology and Gene Therapy, Department of Pharmaceutical and Pharmacological Sciences, Leuven, Belgium. Jan.Derijck@med.kuleuven.be veverka@uochb.cas.cz.
authors.forename: Jan
authors.initials: J
authors.lastname: De Rijck
mesh_terms: D000818:Animals; D001665:Binding Sites; D019427:HIV Integrase / Q000737:chemistry / Q000378:metabolism; D011495:Histone-Lysine N-Methyltransferase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015470:Leukemia, Myeloid, Acute / Q000235:genetics / Q000378:metabolism*; D061986:MCF-7 Cells; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008958:Models, Molecular; D058990:Molecular Targeted Therapy; D051788:Myeloid-Lymphoid Leukemia Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015514:Oncogene Proteins, Fusion / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; C497422:KMT2A protein, human; D015514:Oncogene Proteins, Fusion; C119380:lens epithelium-derived growth factor; D051788:Myeloid-Lymphoid Leukemia Protein; D011495:Histone-Lysine N-Methyltransferase; C497423:Kmt2a protein, mouse; D019427:HIV Integrase