UMMS Affiliation

Department of Molecular, Cell, and Cancer Biology

Publication Date

3-20-2018

Document Type

Article

Disciplines

Amino Acids, Peptides, and Proteins | Biochemistry | Cell Biology | Cells | Fungi | Genetic Phenomena | Investigative Techniques | Laboratory and Basic Science Research | Microbiology | Molecular Biology

Abstract

Nucleosomes 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.

Keywords

Chromatin, Crosslink, Histone, Nucleosome, Protein-protein interaction, Streptavidin affinity chromatography

Rights and Permissions

Copyright Ichikawa and Kaufman. This article is distributed under the terms of the Creative Commons Attribution License (CC BY 4.0).

DOI of Published Version

10.21769/BioProtoc.2770

Source

Bio Protoc. 2018 Mar 20;8(6). doi: 10.21769/BioProtoc.2770. Link to article on publisher's site

Journal/Book/Conference Title

Bio-protocol

Related Resources

Link to Article in PubMed

PubMed ID

29644260

Creative Commons License

Creative Commons Attribution 4.0 License
This work is licensed under a Creative Commons Attribution 4.0 License.

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