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TitleStructural and dynamic mechanisms of CBF3-guided centromeric nucleosome formation.
Journal, issue, pagesNat Commun, Vol. 12, Issue 1, Page 1763, Year 2021
Publish dateMar 19, 2021
AuthorsRuifang Guan / Tengfei Lian / Bing-Rui Zhou / Emily He / Carl Wu / Martin Singleton / Yawen Bai /
PubMed AbstractAccurate chromosome segregation relies on the specific centromeric nucleosome-kinetochore interface. In budding yeast, the centromere CBF3 complex guides the deposition of CENP-A, an H3 variant, to ...Accurate chromosome segregation relies on the specific centromeric nucleosome-kinetochore interface. In budding yeast, the centromere CBF3 complex guides the deposition of CENP-A, an H3 variant, to form the centromeric nucleosome in a DNA sequence-dependent manner. Here, we determine the structures of the centromeric nucleosome containing the native CEN3 DNA and the CBF3core bound to the canonical nucleosome containing an engineered CEN3 DNA. The centromeric nucleosome core structure contains 115 base pair DNA including a CCG motif. The CBF3core specifically recognizes the nucleosomal CCG motif through the Gal4 domain while allosterically altering the DNA conformation. Cryo-EM, modeling, and mutational studies reveal that the CBF3core forms dynamic interactions with core histones H2B and CENP-A in the CEN3 nucleosome. Our results provide insights into the structure of the budding yeast centromeric nucleosome and the mechanism of its assembly, which have implications for analogous processes of human centromeric nucleosome formation.
External linksNat Commun / PubMed:33741944 / PubMed Central
MethodsEM (single particle)
Resolution3.1 - 4.2 Å
Structure data

22696

7k78

EMDB-22696, PDB-7k78:
antibody and nucleosome complex
Method: EM (single particle) / Resolution: 3.1 Å

22697

7k79

EMDB-22697, PDB-7k79:
CBF3
Method: EM (single particle) / Resolution: 4.0 Å

22698

7k7g

EMDB-22698, PDB-7k7g:
nucleosome and Gal4 complex
Method: EM (single particle) / Resolution: 4.2 Å

Chemicals

ChemComp-ZN:
Unknown entry

Source
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
  • saccharomyces cerevisiae (brewer's yeast)
  • mus musculus (house mouse)
KeywordsDNA BINDING PROTEIN/DNA / nucleosome / DNA BINDING PROTEIN / DNA BINDING PROTEIN-DNA complex / CBF3

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