A phospho-dependent mechanism involving NCoR and KMT2D controls a permissive chromatin state at Notch target genes

Nucleic Acids Res. 2016 Jun 2;44(10):4703-20. doi: 10.1093/nar/gkw105. Epub 2016 Feb 23.

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.

MeSH terms

  • Animals
  • Casein Kinase II / metabolism
  • Cell Line
  • Cell Line, Tumor
  • Chromatin / metabolism*
  • Co-Repressor Proteins / metabolism*
  • DNA-Binding Proteins
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism
  • Gene Expression Regulation*
  • Histone Code
  • Histone-Lysine N-Methyltransferase
  • Homeodomain Proteins / chemistry
  • Homeodomain Proteins / metabolism
  • Humans
  • Mice
  • Myeloid-Lymphoid Leukemia Protein / metabolism*
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / metabolism*
  • Phosphorylation
  • Protein Interaction Domains and Motifs
  • RNA-Binding Proteins
  • Receptors, Notch / metabolism*
  • Transcription, Genetic*
  • Xenopus laevis

Substances

  • Chromatin
  • Co-Repressor Proteins
  • DNA-Binding Proteins
  • Drosophila Proteins
  • Homeodomain Proteins
  • Nuclear Proteins
  • RNA-Binding Proteins
  • Receptors, Notch
  • SPEN protein, human
  • Spen protein, mouse
  • Myeloid-Lymphoid Leukemia Protein
  • Histone-Lysine N-Methyltransferase
  • Kmt2b protein, mouse
  • Casein Kinase II