Oral Presentation The 45th Lorne Conference on Protein Structure and Function 2020

CHD4 slides nucleosomes by decoupling entry- and exit-side DNA translocation (#54)

Joel Mackay 1 , Jessica Zhong 1 , Bishnu Paudel 1 , Antoine van Oijen 2 , Daniel Ryan 3 , Jason Low 1
  1. School of Life and Environmental Sciences, University of Sydney, Sydney, NSW, Australia
  2. Molecular Horizons, School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW, Australia
  3. JCSMR, Australian National University, Canberra, ACT, Australia

Any biological process involving the genome – be it transcription, replication or repair – requires the remodelling of histone-DNA interactions. This remodelling is achieved by ATP-dependent DNA translocase enzymes. We set out to determine the mechanism by which one prominent remodeller – the CHD4 – can move histones relative to DNA. We have used single-molecule FRET measurements to demonstrate that the binding energy for CHD4-nucleosome complex formation – even in the absence of nucleotide –triggers significant conformational changes in DNA at the entry side, effectively priming the system for remodelling. During remodelling, flanking DNA enters the nucleosome in a continuous, gradual manner but exits in concerted 4–6 base-pair steps. This decoupling of entry- and exit-side translocation suggests that ATP-driven movement of entry-side DNA builds up strain inside the nucleosome that is subsequently released at the exit side by DNA expulsion. We propose a mechanism for nucleosome sliding based on these and published data.