Interaction of the NuRD corepressor complex with chromatin (360G-Wellcome-207216_Z_17_Z)

Histone deacetylates (HDACs) are important enzymes that play an essential role in cell development and differentiation. HDACs function as part of large protein complexes that are recruited to chromatin to regulate gene expression. The NuRD complex is one of these HDAC-containing complexes, of which the core protein components are known, but the mechanism of recruitment to chromatin is unclear. Our understanding of recruitment to the genome will be enhanced by structural and functional studies on a dimer of NuRD proteins, RBBP4 and MTA1. The MTA1:RBBP4 dimer has been shown to bind to chromatin through parts of the unstructured amino-terminus of histone H3. RBBP4 has also been shown to bind to the transcription factor FOG1 through an amino-terminal motif that is highly conserved in several other transcription factors including BCL11B and SALL1. In this project we propose to purify the MTA1:RBBP4 dimer from mammalian cells and then investigate the tightness of binding to fluorescently labelled peptides using fluorescence anisotropy. We will also set up crystallisation trials to gain structural insight into the mode of binding of these peptides to the MTA1:RBBP4 dimer.