title: The Sequence of Nucleosomal DNA Modulates Sliding by the Chd1 Chromatin Remodeler.
abstract: Chromatin remodelers are ATP-dependent enzymes that are critical for reorganizing and repositioning nucleosomes in concert with many basic cellular processes. For the chromodomain helicase DNA-binding protein 1 (Chd1) remodeler, nucleosome sliding has been shown to depend on the DNA flanking the nucleosome, transcription factor binding at the nucleosome edge, and the presence of the histone H2A/H2B dimer on the entry side. Here, we report that Chd1 is also sensitive to the sequence of DNA within the nucleosome and slides nucleosomes made with the 601 Widom positioning sequence asymmetrically. Kinetic and equilibrium experiments show that poly(dA:dT) tracts perturb remodeling reactions if within one and a half helical turns of superhelix location 2 (SHL2), where the Chd1 ATPase engages nucleosomal DNA. These sequence-dependent effects do not rely on the Chd1 DNA-binding domain and are not due to differences in nucleosome affinity. Using site-specific cross-linking, we show that internal poly(dA:dT) tracts do not block the engagement of the ATPase motor with SHL2, yet they promote multiple translational positions of DNA with respect to both Chd1 and the histone core. We speculate that Chd1 senses the sequence-dependent response of DNA as the remodeler ATPase perturbs the duplex at SHL2. These results suggest that the sequence sensitivity of histones and remodelers occur at unique segments of DNA on the nucleosome, allowing them to work together or in opposition to determine nucleosome positions throughout the genome.
authors.affiliation: Department of Biology, Johns Hopkins University, Baltimore, MD 21218, USA.
authors.forename: Jessica
authors.initials: J
authors.lastname: Winger
authors.affiliation: Department of Biology, Johns Hopkins University, Baltimore, MD 21218, USA; T.C. Jenkins Department of Biophysics, Johns Hopkins University, Baltimore, MD 21218, USA. Electronic address: gdbowman@jhu.edu.
authors.forename: Gregory D
authors.initials: GD
authors.lastname: Bowman
mesh_terms: D002843:Chromatin / Q000378:metabolism*; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
chemical_list: C489847:CHD1 protein, S cerevisiae; D002843:Chromatin; D004268:DNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; D004247:DNA
keywords: Snf2 ATPase; Widom 601 positioning sequence; chromatin remodeling; nucleosome sliding; poly(dA:dT) tracts