Document Type

Article

Publication Date

5-24-2019

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.

Comments

Originally published:
Bing-Rui Zhou, Sathish Yadav, ,K.N., Borgnia, M., Hong, J., Cao, B., Olins, A. L., . . . Zhang, P. (2019). Atomic resolution cryo-EM structure of a native-like CENP-A nucleosome aided by an antibody fragment. Nature Communications, 10, 1-7. doi:http://dx.doi.org.une.idm.oclc.org/10.1038/s41467-019-10247-4

© 2019 The Authors. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”).

Supplementary materials descriptions:
1: Supplementary Figures: 1 – 14, Supplementary Table 1
2: Supplementary Movie: Cryo-EM map and structural features of human CENP-A nucleosome bound to the scFv.

AtomicResolutionCryoEMsupplemental1.pdf (9476 kB)
Supplementary file 1 (.pdf)

AtomicResolutionCryoEMsupplemental2.mp4 (13056 kB)
Supplementary file 2 (.mp4)

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