Investigation of the underlying activities and mechanisms of chromodomain helicase DNA binding protein 4 (CHD4)

Zhong Y1, Paudel B2, Mackay J1 and Van Oijen A2

  1. School of Life and Environmental Science, University of Sydney, Australia.
  2. School of Chemistry, University of Wollongong, Australia.

Nucleosome is the fundamental unit of eukaryotic genome, each of which generally consists of a histone octamer core with 147 bp DNA wrapping around, allowing the whole genome can be efficiently packed into a small nucleus. On the other hand, the presence of nucleosome remodelers ensures the genetic information still remains accessible to regulatory factors. Although it was known for a long time that these remodelers can weaken the interaction between the DNA and histones in an ATP-dependent manner to expose part of the DNA sequence, the underlying mechanisms have not been fully explored, especially for the CHD remodeler family. In the project, we will assemble recombinant nucleosomes and will use them as a substrate to test the remodeling ability of CHD4 and also NuRD (Nucleosome Remodeling Deacetylase) complex, using single molecule FRET (fluorescence resonance energy transfer) experiments. The FRET experiment involves fluorescent labelling of one of the histones and also of the DNA, which allows the monitoring of the translocation of the nucleosome in real time. Other experimental approaches, including bulk experimental measurement of CHD4 remodeling rate in real-time and gel-based remodeling assays, will also be carried out. Together with CryoEM structural studies, these experiments will be used to delineate the biochemical mechanism of remodelling by CHD-family chromatin remodelling proteins.