5ONW

X-Ray crystal structure of a nucleosome core particle with its DNA site-specifically crosslinked to the histone octamer and the two H2A/H2B dimers crosslinked via H2A N38C

Summary for 5ONW

• Entry DOI: 10.2210/pdb5onw/pdb
• Related: 5OMX 5ONG
• Descriptor: DNA (147-MER), Histone H3.2, Histone H4, ... (9 entities in total)
• Functional Keywords: nucleosome core particle, histones, disulfide, convertible nucleotide, dna, dna binding protein
• Biological source: Homo sapiens (Human); More
• Cellular location: Nucleus: P84233 P62799 P02281; Chromosome . Nucleus : Q6AZJ8
• Total number of polymer chains: 10
• Total formula weight: 200300.39

Authors

Frouws, T.D.,Barth, P.D.,Richmond, T.J. (deposition date: 2017-08-04, release date: 2017-11-22, Last modification date: 2024-11-06)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (2.8 Å)