5ONG

X-Ray crystal structure of a nucleosome core particle with its DNA site-specifically crosslinked to the histone octamer

5ONG の概要

• エントリーDOI: 10.2210/pdb5ong/pdb
• 関連するPDBエントリー: 5OMX
• 分子名称: DNA (147-MER), Histone H3.2, Histone H4, ... (9 entities in total)
• 機能のキーワード: nucleosome core particle, histones, disulfide, convertible nucleotide, dna, dna binding protein
• 由来する生物種: Homo sapiens (Human); 詳細
• 細胞内の位置: Nucleus: P84233 P62799 P02281; Chromosome . Nucleus : Q6AZJ8
• タンパク質・核酸の鎖数: 10
• 化学式量合計: 200322.31

構造登録者

Frouws, T.D.,Barth, P.D.,Richmond, T.J. (登録日: 2017-08-03, 公開日: 2017-11-22, 最終更新日: 2024-11-13)

主引用文献

Frouws, T.D.,Barth, P.D.,Richmond, T.J.
Site-Specific Disulfide Crosslinked Nucleosomes with Enhanced Stability.
J. Mol. Biol., 430:45-57, 2018

PubMed Abstract: We engineered nucleosome core particles (NCPs) with two site-specific cysteine crosslinks that increase the stability of the particle. The first disulfide was introduced between the two copies of H2A via an H2A-N38C point mutation, effectively crosslinking the two H2A/H2B heterodimers together to stabilize the histone octamer against H2A/H2B dimer dissociation. The second crosslink was engineered between an R40C point mutation on the N-terminal tail of H3 and the NCP DNA ends by the introduction of a convertible nucleotide. This crosslink maintains the nucleosome DNA in a fixed translational setting relative to the histone octamer and prevents dilution-driven dissociation. The X-ray crystal structures of NCPs containing the disulfides in isolation and in combination were determined. Both disulfides stabilize the structure of the NCP without disturbing the overall structure. Nucleosomes containing these modifications will be advantageous for biochemical and structural studies as a consequence of their greater resistance to dissociation during high dilution in purification, elevated salt for crystallization and vitrification for cryogenic electron microscopy.

PubMed: 29113904
DOI: 10.1016/j.jmb.2017.10.029

実験手法

X-RAY DIFFRACTION (2.797 Å)