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Biochemical Analysis of Dimethyl Suberimidate-crosslinked Yeast Nucleosomes
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-ter...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Bio-Protocol
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5891137/ https://www.ncbi.nlm.nih.gov/pubmed/29644260 http://dx.doi.org/10.21769/BioProtoc.2770 |
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author | Ichiakwa, Yuichi Kaufman, Paul D. |
author_facet | Ichiakwa, Yuichi Kaufman, Paul D. |
author_sort | Ichiakwa, Yuichi |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-5891137 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Bio-Protocol |
record_format | MEDLINE/PubMed |
spelling | pubmed-58911372018-04-09 Biochemical Analysis of Dimethyl Suberimidate-crosslinked Yeast Nucleosomes Ichiakwa, Yuichi Kaufman, Paul D. Bio Protoc Methods Article 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. Bio-Protocol 2018-03-20 /pmc/articles/PMC5891137/ /pubmed/29644260 http://dx.doi.org/10.21769/BioProtoc.2770 Text en ©Copyright Ichiakwa and Kaufman. https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (CC BY 4.0). |
spellingShingle | Methods Article Ichiakwa, Yuichi Kaufman, Paul D. Biochemical Analysis of Dimethyl Suberimidate-crosslinked Yeast Nucleosomes |
title | Biochemical Analysis of Dimethyl Suberimidate-crosslinked Yeast Nucleosomes |
title_full | Biochemical Analysis of Dimethyl Suberimidate-crosslinked Yeast Nucleosomes |
title_fullStr | Biochemical Analysis of Dimethyl Suberimidate-crosslinked Yeast Nucleosomes |
title_full_unstemmed | Biochemical Analysis of Dimethyl Suberimidate-crosslinked Yeast Nucleosomes |
title_short | Biochemical Analysis of Dimethyl Suberimidate-crosslinked Yeast Nucleosomes |
title_sort | biochemical analysis of dimethyl suberimidate-crosslinked yeast nucleosomes |
topic | Methods Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5891137/ https://www.ncbi.nlm.nih.gov/pubmed/29644260 http://dx.doi.org/10.21769/BioProtoc.2770 |
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