UMMS Affiliation
Department of Molecular, Cell, and Cancer Biology
Publication Date
2018-03-20
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
PubMed ID
29644260
Repository Citation
Ichikawa Y, Kaufman PD. (2018). Biochemical Analysis of Dimethyl Suberimidate-crosslinked Yeast Nucleosomes. Open Access Articles. https://doi.org/10.21769/BioProtoc.2770. Retrieved from https://escholarship.umassmed.edu/oapubs/3403
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.
Included in
Amino Acids, Peptides, and Proteins Commons, Biochemistry Commons, Cell Biology Commons, Cells Commons, Fungi Commons, Genetic Phenomena Commons, Investigative Techniques Commons, Laboratory and Basic Science Research Commons, Microbiology Commons, Molecular Biology Commons