Biochemical Analysis of Dimethyl Suberimidate-crosslinked Yeast Nucleosomes
UMass Chan AffiliationsDepartment of Molecular, Cell, and Cancer Biology
Streptavidin affinity chromatography
Amino Acids, Peptides, and Proteins
Laboratory and Basic Science Research
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AbstractNucleosomes are the fundamental unit of eukaryotic chromosome packaging, comprised of 147 bp of DNA wrapped around two molecules of each of the core histone proteins H2A, H2B, H3, and H4. Nucleosomes are symmetrical, with one axis of symmetry centered on the homodimeric interaction between the C-termini of the H3 molecules. To explore the functional consequences of nucleosome symmetry, we designed an obligate pair of H3 heterodimers, termed H3X and H3Y, allowing us to compare cells with single or double H3 alterations. Our biochemical validation of the heterodimeric X-Y interaction included intra-nucleosomal H3 crosslinking using dimethyl suberimidate (DMS). Here, we provide a detailed protocol for the use of DMS to analyze yeast nucleosomes.
Bio Protoc. 2018 Mar 20;8(6). doi: 10.21769/BioProtoc.2770. Link to article on publisher's site
Permanent Link to this Itemhttp://hdl.handle.net/20.500.14038/40599
RightsCopyright Ichikawa and Kaufman. This article is distributed under the terms of the Creative Commons Attribution License (CC BY 4.0).
Except where otherwise noted, this item's license is described as Copyright Ichikawa and Kaufman. This article is distributed under the terms of the Creative Commons Attribution License (CC BY 4.0).