Atomic resolution cryo-EM structure of a native-like CENP-A nucleosome aided by an antibody fragment
By Bing-Rui Zhou, K. Yadav, M. Borgnia, Jingjun Hong, B. Cao, A. Olins, D. Olins, Yawen Bai, Ping Zhang. 2019
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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. CENP-A histone variants replace histones H3 at centromeres. Here the authors use a single-chain antibody fragment (scFv) to stabilize human CENP-A nucleosome containing a native α-satellite DNA and solved its structure by cryo-EM to 2.6 Å resolution, providing insight into the structure and function of the CENP-A nucleosome.
Subfields: Molecular Biology, Structural Biology
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- Distinct Structures and Dynamics of Chromatosomes with Different Human Linker Histone Isoforms (2020, methodology): “We have previously shown that a single-chain antibody fragment (scFv) that binds to the core histones can prevent the nucleosome from dissociating during the blotting-freezing process of sample preparation for single-particle cryo-EM imaging, allowing determination of the nucleosome structures with ”