title: A phospho-dependent mechanism involving NCoR and KMT2D controls a permissive chromatin state at Notch target genes.
abstract: The transcriptional shift from repression to activation of target genes is crucial for the fidelity of Notch responses through incompletely understood mechanisms that likely involve chromatin-based control. To activate silenced genes, repressive chromatin marks are removed and active marks must be acquired. Histone H3 lysine-4 (H3K4) demethylases are key chromatin modifiers that establish the repressive chromatin state at Notch target genes. However, the counteracting histone methyltransferase required for the active chromatin state remained elusive. Here, we show that the RBP-J interacting factor SHARP is not only able to interact with the NCoR corepressor complex, but also with the H3K4 methyltransferase KMT2D coactivator complex. KMT2D and NCoR compete for the C-terminal SPOC-domain of SHARP. We reveal that the SPOC-domain exclusively binds to phosphorylated NCoR. The balance between NCoR and KMT2D binding is shifted upon mutating the phosphorylation sites of NCoR or upon inhibition of the NCoR kinase CK2β. Furthermore, we show that the homologs of SHARP and KMT2D in Drosophila also physically interact and control Notch-mediated functions in vivo Together, our findings reveal how signaling can fine-tune a committed chromatin state by phosphorylation of a pivotal chromatin-modifier.
authors.affiliation: University Medical Center Ulm, Center for Internal Medicine, Department of Internal Medicine I, Albert-Einstein-Allee 23, 89081 Ulm, Germany franz.oswald@uni-ulm.de.
authors.forename: Franz
authors.initials: F
authors.lastname: Oswald
authors.affiliation: Swiss Institute for Experimental Cancer Research, Lausanne, Switzerland.
authors.forename: Patrick
authors.initials: P
authors.lastname: Rodriguez
authors.affiliation: Institute of Biochemistry, University of Giessen, Friedrichstrasse 24, 35392 Giessen, Germany Spemann Graduate School of Biology and Medicine (SGBM), Faculty of Biology, Albert Ludwigs University Freiburg, Germany.
authors.forename: Benedetto Daniele
authors.initials: BD
authors.lastname: Giaimo
authors.affiliation: Instituto de Neurociencias, Consejo Superior de Investigaciones Cientificas-Universidad Miguel Hernández, Campus de Sant Joan, Alicante, Spain.
authors.forename: Zeus A
authors.initials: ZA
authors.lastname: Antonello
authors.affiliation: Instituto de Neurociencias, Consejo Superior de Investigaciones Cientificas-Universidad Miguel Hernández, Campus de Sant Joan, Alicante, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Mira
authors.affiliation: Max-Planck-Institute of Immunobiology and Epigenetics, Stübeweg 51, 79108 Freiburg, Germany.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Mittler
authors.affiliation: University Medical Center Ulm, Center for Internal Medicine, Department of Internal Medicine I, Albert-Einstein-Allee 23, 89081 Ulm, Germany.
authors.forename: Verena N
authors.initials: VN
authors.lastname: Thiel
authors.affiliation: Department of Molecular Genetics, Biochemistry, and Microbiology, University of Cincinnati, Cincinnati, OH 45267, USA.
authors.forename: Kelly J
authors.initials: KJ
authors.lastname: Collins
authors.affiliation: Department of Molecular Genetics, Biochemistry, and Microbiology, University of Cincinnati, Cincinnati, OH 45267, USA.
authors.forename: Nassif
authors.initials: N
authors.lastname: Tabaja
authors.affiliation: Institute for Biochemistry and Molecular Biology, Ulm University, Albert-Einstein-Allee 11, 89081 Ulm, Germany International Graduate School in Molecular Medicine Ulm (IGradU), Ulm University, Albert-Einstein-Allee 11, 89081 Ulm, Germany.
authors.forename: Wiebke
authors.initials: W
authors.lastname: Cizelsky
authors.affiliation: Institute for Biochemistry and Molecular Biology, Ulm University, Albert-Einstein-Allee 11, 89081 Ulm, Germany International Graduate School in Molecular Medicine Ulm (IGradU), Ulm University, Albert-Einstein-Allee 11, 89081 Ulm, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Rothe
authors.affiliation: Institute for Biochemistry and Molecular Biology, Ulm University, Albert-Einstein-Allee 11, 89081 Ulm, Germany.
authors.forename: Susanne J
authors.initials: SJ
authors.lastname: Kühl
authors.affiliation: Institute for Biochemistry and Molecular Biology, Ulm University, Albert-Einstein-Allee 11, 89081 Ulm, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Kühl
authors.affiliation: Institute of Biochemistry, University of Giessen, Friedrichstrasse 24, 35392 Giessen, Germany.
authors.forename: Francesca
authors.initials: F
authors.lastname: Ferrante
authors.affiliation: Institute of Biochemistry, University of Giessen, Friedrichstrasse 24, 35392 Giessen, Germany.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Hein
authors.affiliation: Department of Molecular Genetics, Biochemistry, and Microbiology, University of Cincinnati, Cincinnati, OH 45267, USA.
authors.forename: Rhett A
authors.initials: RA
authors.lastname: Kovall
authors.affiliation: Instituto de Neurociencias, Consejo Superior de Investigaciones Cientificas-Universidad Miguel Hernández, Campus de Sant Joan, Alicante, Spain.
authors.forename: Maria
authors.initials: M
authors.lastname: Dominguez
authors.affiliation: Institute of Biochemistry, University of Giessen, Friedrichstrasse 24, 35392 Giessen, Germany Tilman.Borggrefe@biochemie.med.uni-giessen.de.
authors.forename: Tilman
authors.initials: T
authors.lastname: Borggrefe
mesh_terms: D000818:Animals; D047390:Casein Kinase II / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D002843:Chromatin / Q000378:metabolism*; D056970:Co-Repressor Proteins / Q000378:metabolism*; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D042421:Histone Code; D011495:Histone-Lysine N-Methyltransferase; D018398:Homeodomain Proteins / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D051379:Mice; D051788:Myeloid-Lymphoid Leukemia Protein / Q000378:metabolism*; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D054730:Protein Interaction Domains and Motifs; D016601:RNA-Binding Proteins; D051880:Receptors, Notch / Q000378:metabolism*; D014158:Transcription, Genetic; D014982:Xenopus laevis
chemical_list: D002843:Chromatin; D056970:Co-Repressor Proteins; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; D051880:Receptors, Notch; C476510:SPEN protein, human; C121253:Spen protein, mouse; D051788:Myeloid-Lymphoid Leukemia Protein; D011495:Histone-Lysine N-Methyltransferase; C510860:Kmt2d protein, mouse; D047390:Casein Kinase II