title: The PHD3 domain of MLL acts as a CYP33-regulated switch between MLL-mediated activation and repression .
abstract: The mixed lineage leukemia (MLL) gene plays a critical role in epigenetic regulation of gene expression and is a frequent target of chromosomal translocations leading to leukemia. MLL plant homeodomain 3 (PHD3) is lost in all MLL translocation products, and reinsertion of PHD3 into MLL fusion proteins abrogates their transforming activity. PHD3 has been shown to interact with the RNA-recognition motif (RRM) domain of human nuclear Cyclophilin33 (CYP33). Here, we show that CYP33 mediates downregulation of the expression of MLL target genes HOXC8, HOXA9, CDKN1B, and C-MYC, in a proline isomerase-dependent manner. This downregulation correlates with the reduction of trimethylated lysine 4 of histone H3 (H3K4me3) and histone H3 acetylation. We have structurally characterized both the PHD3 and CYP33 RRM domains and analyzed their binding to one another. The PHD3 domain binds H3K4me3 (preferentially) and the CYP33 RRM domain at distinct sites. Our binding data show that binding of H3K4me3 to PHD3 and binding of the CYP33 RRM domain to PHD3 are mutually inhibitory, implying that PHD3 is a molecular switch for the transition between activation and repression of target genes. To explore the possible mechanism of CYP33/PHD3-mediated repression, we have analyzed the CYP33 proline isomerase activity on various H3 and H4 peptides and shown selectivity for two sites in H3. Our results provide a possible mechanism for the MLL PHD3 domain to act as a switch between activation and repression.
authors.affiliation: Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, Virginia 22908, USA.
authors.forename: Sangho
authors.initials: S
authors.lastname: Park
authors.forename: Ute
authors.initials: U
authors.lastname: Osmers
authors.forename: Gayathree
authors.initials: G
authors.lastname: Raman
authors.forename: Rebecca H
authors.initials: RH
authors.lastname: Schwantes
authors.forename: Manuel O
authors.initials: MO
authors.lastname: Diaz
authors.forename: John H
authors.initials: JH
authors.lastname: Bushweller
mesh_terms: D001665:Binding Sites; D021983:Cyclophilins / Q000502:physiology*; D015536:Down-Regulation; D005786:Gene Expression Regulation; D006657:Histones / Q000378:metabolism; D006801:Humans; D051788:Myeloid-Lymphoid Leukemia Protein / Q000737:chemistry / Q000502:physiology*; D019696:Peptidylprolyl Isomerase / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012097:Repressor Proteins; D015854:Up-Regulation
chemical_list: D006657:Histones; D012097:Repressor Proteins; D051788:Myeloid-Lymphoid Leukemia Protein; D021983:Cyclophilins; C114522:PPIE protein, human; D019696:Peptidylprolyl Isomerase