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-Structure paper
Title | Atomic resolution cryo-EM structure of a native-like CENP-A nucleosome aided by an antibody fragment. |
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Journal, issue, pages | Nat Commun, Vol. 10, Issue 1, Page 2301, Year 2019 |
Publish date | May 24, 2019 |
Authors | Bing-Rui Zhou / K N Sathish Yadav / Mario Borgnia / Jingjun Hong / Baohua Cao / Ada L Olins / Donald E Olins / Yawen Bai / Ping Zhang / |
PubMed 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 ...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. |
External links | Nat Commun / PubMed:31127102 / PubMed Central |
Methods | EM (single particle) |
Resolution | 2.6 - 3.395 Å |
Structure data | EMDB-0586, PDB-6o1d: EMDB-8938, PDB-6dzt: |
Source |
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Keywords | NUCLEAR PROTEIN / Nucleosome / Single chain antibody / charge-charge interaction / acidic patch. / CENP-A / Centromere / Widom's 601 DNA / anti-nucleosome antibody / acidic patch / Native alpha satellite DNA |