Atomic resolution cryo-EM structure of a native-like CENP-A nucleosome aided by an antibody fragment.
Nat Commun
; 10(1): 2301, 2019 05 24.
Article
em En
| MEDLINE
| ID: mdl-31127102
ABSTRACT
Genomic DNA in eukaryotes is organized into chromatin through association with core histones to form nucleosomes, each distinguished by their DNA sequences and histone variants. Here, we used a single-chain antibody fragment (scFv) derived from the anti-nucleosome antibody mAb PL2-6 to stabilize human CENP-A nucleosome containing a native α-satellite DNA and solved its structure by the cryo-electron microscopy (cryo-EM) to 2.6 Å resolution. In comparison, the corresponding cryo-EM structure of the free CENP-A nucleosome could only reach 3.4 Å resolution. We find that scFv binds to a conserved acidic patch on the histone H2A-H2B dimer without perturbing the nucleosome structure. Our results provide an atomic resolution cryo-EM structure of a nucleosome and insight into the structure and function of the CENP-A nucleosome. The scFv approach is applicable to the structural determination of other native-like nucleosomes with distinct DNA sequences.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
DNA Satélite
/
Nucleossomos
/
Proteína Centromérica A
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article