Biophysical Characterization of Chromatin Remodeling Protein CHD4.
Chromatin-remodeling ATPases modulate histones-DNA interactions within nucleosomes and regulate transcription. At the heart of remodeling, ATPase is a helicase-like motor flanked by a variety of conserved targeting domains. CHD4 is the core subunit of the nucleosome remodeling and deacetylase complex NuRD and harbors tandem plant homeo finger (tPHD) and chromo (tCHD) domains. We describe a multifaceted approach to link the domain structure with function, using quantitative assays for DNA and histone binding, ATPase activity, shape reconstruction from solution scattering data, and single molecule translocation assays. These approaches are complementary to high-resolution structure determination.
Faculty of Life Sciences, Manchester Institute of Biotechnology, University of Manchester, Manchester, UK.
Rosa
R
Morra
Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK.
Tomas
T
Fessl
Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK.
Yuchong
Y
Wang
School of Life Sciences, University of Sussex, Brighton, UK.
Erika J
EJ
Mancini
Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK. R.Tuma@leeds.ac.uk.
Roman
R
Tuma
D001324:Autoantigens / Q000737:chemistry* / Q000378:metabolism*; D055592:Biophysical Phenomena; D004247:DNA / Q000378:metabolism; D000067493:Dynamic Light Scattering; D006657:Histones / Q000378:metabolism; D006801:Humans; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex / Q000737:chemistry* / Q000378:metabolism*; D000072417:Protein Domains; D021381:Protein Transport; D000072760:Single Molecule Imaging
D001324:Autoantigens; C115323:CHD4 protein, human; D006657:Histones; D004247:DNA; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex
ATPase; FRET; Nucleosome; SAXS; Surface plasmon resonance; TIRF