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TitleA method for cryo-EM analysis of eukaryotic nucleosomes reconstituted in bacterial cells.
Journal, issue, pagesiScience, Vol. 29, Issue 1, Page 114453, Year 2026
Publish dateJan 16, 2026
AuthorsCheng-Han Ho / Yuki Kobayashi / Mitsuo Ogasawara / Yoshimasa Takizawa / Hitoshi Kurumizaka /
PubMed AbstractConventional methods for preparing nucleosomes are time-consuming and technically demanding. In the present study, we extended the approach of generating nucleosomes in by the co-expression of all ...Conventional methods for preparing nucleosomes are time-consuming and technically demanding. In the present study, we extended the approach of generating nucleosomes in by the co-expression of all four histones, allowing nucleosomes to be assembled in cells. The bacterially reconstituted nucleosomes can be readily prepared from the cells and directly subjected to cryo-EM single particle analysis. Using this method, we obtained a 2.56 Å nucleosome structure that is highly similar to a previously reported nucleosome crystal structure, validating the use of nucleosomes formed in for cryo-EM analysis. Unexpectedly, we also discovered a non-canonical nucleosome structure, in which two hexasomes are closely packed. This system provides a robust and efficient platform for structural studies of nucleosomes and nucleosome variants, and may facilitate the discovery of chromatin architectures.
External linksiScience / PubMed:41541688 / PubMed Central
MethodsEM (single particle)
Resolution4.94 Å
Structure data

65716

9w74

EMDB-65716, PDB-9w74:
Cryo-EM structure of the close-packed di-hexasome (CPDH)
Method: EM (single particle) / Resolution: 4.94 Å

Source
  • xenopus laevis (African clawed frog)
  • Mus musculus (house mouse)
  • escherichia coli (E. coli)
KeywordsGENE REGULATION / Chromatin / Nucleosome / Hexasome / Subnucleosome / Histone / DNA

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